TMP D19 C

Chemical Senses Advance Access published August 1, 2013
Chem. Senses doi:10.1093/chemse/bjt036
ABSTRACTS
Abstracts from the 35th Annual Meeting of AChemS
#1 GIVAUDAN LECTURE: NON-MODEL MODELS Recent advances in the structural biology of G protein-cou-
IN OLFACTION pled receptors have helped to unravel the intricacies of ligand
binding. Similarly structural and biochemical analyses of het-
erotrimeric G proteins have affirmed our understanding of the
Bill S. Hansson
mechanism underlying effector interactions and GTPase activ-
Max Planck Institute for Chemical Ecology, Jena, Germany ity. The recent crystal structure of a prototypic GPCR, the beta2
-adrenergic receptor (beta2 AR), in a complex with the stimu-
Our knowledge regarding olfactory structure and function latory G protein, Gs, trapped in its nucleotide-free state, has
has taken a quantum leap since the characterization of puta-
Downloaded from http://chemse.oxfordjournals.org/ by Sergey Novikov on October 17, 2013

now provided models for activation of G proteins by GPCRs.
tive olfactory receptors both in vertebrates and insects. Still, These data have helped to delineate how hormone binding to
the understanding of both ecological and evolutionary pro- GPCRs leads to GDP release on G proteins, the principle step
cesses is lagging behind. What do different odors really mean that precedes GTP binding and G protein activation. The crys-
to animals, and how has the system evolved to allow them to tal structure, together with data from single particle reconstruc-
respond behaviorally in a relevant manner. To understand tions by electron microscopy and deuterium exchange mass
these topics better we use both a model insect (Drosophila spectrometry, reveal dramatic changes in the G protein alpha-
melanogaster), and a number of non-model arthropods subunit. The structural data also suggest that G proteins may
(moths, ants, primitive insects, land-living crustaceans). allosterically regulate the receptor by stabilizing a closed con-
I will spend a short introductory part on recent progress in formation on the extracellular face of the receptor. Radioligand
our work on drosophilid flies, touching on investigations of binding analyses suggest that G protein coupling slows ligand
hard-wired smell-driven behavior. The rest of my talk I will dissociation, consistent with the observed structural changes in
devote to non-model arthropods. the extracellular face. Such structural changes account for the
In the desert ant, Cataglyphis fortis, we have built on the clas- slower observed ligand dissociation rates and likely account for
sic investigations by the group of Rüdiger Wehner in Zürich. G protein-dependent high affinity agonist binding. Together
They demonstrated the amazing ability of these tiny insects to these data support a plausible model for the mechanism for
find their way home in a salt desert using vector-based orienta- receptor-mediated nucleotide exchange, G protein activation
tion. In our investigations we show how the ants use odor cues and agonist binding. Acknowledgements: GM083118
to pinpoint their nest entrance, but how this orientation is, in a
life-saving fashion, weighed against vector orientation. We also
show how the ants use olfactory cues to find their main source #3 SYMPOSIUM: THE STRUCTURAL BASIS OF
of food; dead insects, in a highly efficient way, and how this CHEMOSENSORY SIGNALING
smell-based food search is independent of the home vector.
To investigate the evolution of arthropod olfactory systems Structural Determinants of TRPV Channel Activation and
we make use of both highly primitive, archeopteran insects (in Desensitization
comparison to their neopteran relatives), and of land-living Rachelle Gaudet
crustaceans that have entered the terrestrial environment dur-
Harvard University Cambridge, MA, USA
ing the last five million years. Using a combination of tran-
scriptomics, electrophysiology, morphology and bioassays we My lab is broadly interested in the mechanisms of
show that the neopteran divide coincides with a drastic devel- signaling and transport across cellular membranes. Much
opment in the insect olfactory system, and that some land- of our research centers on TRP channels and their role in
living crustaceans have developed enormous central olfactory sensory perception. We focus on temperature-sensitive ion
systems, while others seem to have abandoned olfaction. channels, particularly TRPV1 and TRPA1. TRP channels
are challenging structural biology targets because they are
large multidomain eukaryotic membrane proteins and are
#2 SYMPOSIUM: THE STRUCTURAL BASIS OF not naturally abundant. We take complementary approaches
CHEMOSENSORY SIGNALING to obtain structural and functional information on TRP
channels. One strategy is to divide and conquer: determine
G protein coupling GPCRs: structural and functional insights
crystal structures of isolated domains of TRP channels. The
into reciprocal G protein and GPCR interactions
results can then combined with genetic, biochemical and
Roger K. Sunahara physiological data to advance our understanding of TRP
University of Michigan Medical School Ann Arbor, MI, USA channel function. It also provides valuable insights as we
© The Author 2013. Published by Oxford University Press. All rights reserved.
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Page 2 of 124 Abstracts
tackle the structure determination of whole TRP channels. every amino acid in the receptor and screened each mutant’s
Acknowledgements: NIH grant R01GM081340 to RG. functional activity in response to unique ligands, both agonists
and inhibitors, using a Ca2+-flux signaling assay. We identified
#4 SYMPOSIUM: THE STRUCTURAL BASIS OF amino acids that, when substituted, abrogate signaling for all
CHEMOSENSORY SIGNALING agonists, as well as residues important for the activity of only
specific ligands. These critical residues define both the binding
Atomistic Structures of human olfactory and taste receptors sites for various TAS2R16 ligands as well as motifs important
William A Goddard, Soo-Kyung Kim for signal transduction, and will help guide the structural under-
standing of bitter taste receptor function. Acknowledgements:
Ravinder Abrol Materials and Process Simulation Center,
NIH DC002995 NIH DC010105 NIH GM072379
California Instute of Technology Pasadena, CA, USA
Olfactory receptors (ORs) responsible for mediating the sense #7 INDUSTRY SYMPOSIUM: TASTE AND SMELL
of smell are G protein-coupled receptors (GPCRs). Through IN TRANSLATION: APPLICATIONS FROM BASIC
differential interactions the ~400 ORs allow us to recognize
RESEARCH
countless ligands. However little is known about the specific
interactions responsible. We use first principles (non homology) Individual differences in oral fat detection and their health
methods to predict the ensemble of 24 low energy structures implications
for the human OR1G1 (hOR1G1) using the GEnSeMBLE Richard Mattes
(GPCR Ensemble of Structures in Membrane BiLayer
Environment) method. The final best structure has strong Purdue University, West Lafayette IN 47907 USA
interhelical H-bonding networks in TMs 1-2-7 (N1.50, D2.50 The fat content of foods and the oral perception of this
and N7.49), and TMs 2–4 (N2.45 and W4.50), which have been fat influence food choice and a wide range of physiological
observed in x-ray and predicted structures for some members responses with health implications. Triacylglycerol, the
of the class A (rhodopsin family) GPCRs. We also find a salt- predominant form of dietary fat, generally contributes
bridge between the conserved D3.49 and K6.30 in the D(E)RY positive somatosensory attributes to foods and are viewed as
region, associated with the inactive form for class A GPCRs. orexigenic. In contrast, non-esterified fatty acids, likely taste
In addition, a hOR specific interaction was formed between stimuli, typically elicit negative hedonic responses and are
the conserved D/E3.39 and H6.40 in hORs, which might be an anorexigenic. The latter also exerts unique effects on cephalic,
important conformational constraint in all hORs since they are intestinal and post-absorptive lipid metabolism. However,
highly conserved (~94 and 98% respectively) among all hORs. individual variability in taste and physiological responses
We expect our predicted 3D structures for hOR1G1 to provide is extremely high raising question about distinct groups of
a guide in understanding the structural patterns for all hORs. fat tasters/responder This presentation will examine possible
This might help design better odorants for foods and perfumes sources of individual variability such as learning, BMI status,
and improved drugs for neurodegenerative disorders, such as age, gender and stimuli properties to facilitate better study
anosmia (inability to detect odors) or hyposmia (decreased abil- designs and interpretation of the functions of “fat taste.”
ity to detect odors). The structure and conserved elements of
the hORs are compared to our recent prediction of the atomis- #8 INDUSTRY SYMPOSIUM: TASTE AND SMELL
tic structure for the human TAS2R38 Bitter Receptor. IN TRANSLATION: APPLICATIONS FROM BASIC
RESEARCH
#5 SYMPOSIUM: THE STRUCTURAL BASIS OF
Mechanisms for Fat Taste
CHEMOSENSORY SIGNALING
Timothy Gilbertson
Comprehensive mapping of functional sites for agonists
and inhibitors of the bitter taste receptor TAS2R16 Department of Biology, Utah State University, Logan UT
84322 USA
Joseph B. Rucker
It has been well over a decade since our laboratory first iden-
Integral Molecular Philadelphia, PA, USA
tified the ability of free fatty acids to activate mammalian
Bitter tastes are detected at the cellular level by a diverse family taste receptors cells, consistent with there being a “taste of
of taste receptors (TAS2Rs) belonging to the G protein-coupled fat”. Since that time, the ability of fatty acids to act as the
receptor (GPCR) superfamily. The lack of direct structural proximate stimuli for fat taste has been validated in a number
information for TAS2Rs (and most GPCRs) limits our under- of species spanning the molecular, cellular and behavioral
standing of their structural features responsible for ligand bind- levels. We have recently identified several novel fatty acid-
ing and signaling. Using a high-throughput approach called activated G protein-coupled receptors that, in conjunction
Shotgun Mutagenesis Mapping, we created a comprehensive with the fatty acid binding protein CD36, allow the recogni-
TAS2R16 mutation library with defined point mutations at tion of chemically distinct classes of free fatty acids. This
Abstracts Page 3 of 124
presentation will summarize what is known about the recep- increased postprandial PYY and GLP-1 favouring enhanced
tors and transduction pathway for free fatty acids. satiety. An early and exaggerated insulin response mediates
improved glycaemic control. The rodent model of bypass
#9 INDUSTRY SYMPOSIUM: TASTE AND SMELL showed elevated PYY, GLP-1 and gut hypertrophy compared
IN TRANSLATION: APPLICATIONS FROM BASIC with sham-operated rats. Moreover, exogenous PYY reduced
RESEARCH food intake while blockade of endogenous PYY increased
food intake. A prospective follow-up human study of gastric
Insights from olfactory receptor screening bypass showed progressively increasing PYY, enteroglucagon,
Joel Mainland and GLP-1 responses associated with enhanced satiety.
Blocking these responses in animal and human models leads
Monell Chemical Senses Center, Philadelphia PA 19104 USA
to increased food intake. Thus, following gastric bypass, a
A major advances in the taste field in recent decades was the pleiotrophic endocrine response may contribute to improved
identification of receptors that mediate taste. Expression of these glycaemic control, appetite reduction, and long-term lowering
receptors in heterologous cell-based assays has allowed scientists of body weight. We have shown that changes occur in the
in both academia and industry to screen for novel taste agonists, sensory, reward and physiological domains of taste that
antagonists, and modifiers. Similar studies in olfactory receptors may mechanistically contribute to the alterations in food
have lagged behind the taste field due to the size of the receptor preferences after gastric bypass. The sustained nature of weight
family as well as difficulties in expressing the receptors in heter- loss, reduced appetite and shifts in food preferences may be
ologous cells. In this talk we will explore the current state of the explained by gut adaptation and chronic hormone elevation.
science in olfactory receptor screening, relationships between
odorant structure and odor quality, and the identification of
agonists, antagonists and modifiers for olfactory receptors. #13 PRESIDENTIAL SYMPOSIUM: GUT PEPTIDE
INTERACTIONS BETWEEN TASTE, FEEDING,
#10 INDUSTRY SYMPOSIUM: TASTE AND SMELL AND REWARD
IN TRANSLATION: APPLICATIONS FROM BASIC
Common Mechanisms of Alimentary Chemosensation:
RESEARCH
Implications for Taste, Ingestion and Glucose Homeostasis
Mechanisms of olfactory adaptation
Steven D. Munger1,2
Haiqing Zhao 1
University of Maryland School of Medicine, Department of
Johns Hopkins University, Baltimore MD 21218 USA Anatomy and Neurobiology Baltimore, MD, USA, 2University
of Maryland School of Medicine, Department of Medicine
Olfactory receptor cells exhibit reduced sensitivity upon pro-
Baltimore, MD, USA
longed or repeated odor exposure––a phenomenon known
as adaptation. Adaptation at the cellular level is thought to The last two decades has seen a growing recognition that a
underlie, at least in part, the perceptual desensitization of common molecular toolkit is employed along the length of
an odor over time. In vertebrates, several calcium-dependent the alimentary canal to detect and respond to nutrients. For
feedback mechanisms have been proposed to account for example, many of the same proteins that are critical for rec-
adaption of olfactory receptor cells. Recent studies using ognizing sweet, bitter and umami taste stimuli in the mouth,
molecular genetic approaches that allow selective disruption including T1R and T2R taste receptors and the transduc-
of these calcium-dependent mechanisms have provide new tion proteins α-gustducin and TRPM5, are found through-
insight into how olfactory adaptation may occur. out the gastrointestinal (GI) tract and associated organs.
Similarly, taste buds express a number of neuropeptides that
#12 PRESIDENTIAL SYMPOSIUM: GUT PEPTIDE are perhaps best understood in other systems as endocrine
INTERACTIONS BETWEEN TASTE, FEEDING, factors that impact nutrient metabolism and/or ingestive
AND REWARD behaviors. A better understanding of the molecular mecha-
nisms that couple nutrient detection to peptide secretion in
Bariatric surgery and appetite gustatory and GI tissues could lead to the identification of
Carel Le Roux new pharmacological targets for impacting ingestion, sati-
ety, nutrient assimilation or glycemic control. I will discuss
Experimental Pathology, UCD Conway Institute, School of
our recent studies in rodents, including animals deficient in
Medicine and Medical Science, University College Dublin,
specific taste receptor subunits or receiving Roux-en-Y gas-
Ireland
tric bypass, that provide new insights into the mechanisms
A good model to investigate appetite reduction in humans of nutrient response in the mouth and gut and the role
and rodents with associated major weight loss is bariatric of these mechanisms in taste coding, post-ingestive nutri-
surgery. Gastric bypass, but not gastric banding caused ent response, and the regulation of glucose homeostasis.
Acknowledgements: NIDCD (DC010110), Tate & Lyle While lateral inhibition between functional processing units
Americas, Ajinomoto Amino Acid Research Program is a fundamental feature of sensory systems, the organiza-
tion of lateral inhibitory circuits in the olfactory system
#14 PRESIDENTIAL SYMPOSIUM: GUT PEPTIDE and their impact on odor representations remain unclear.
Here, we addressed this question in the olfactory bulb (OB)
INTERACTIONS BETWEEN TASTE, FEEDING,
using in vivo two-photon imaging from genetically-defined
AND REWARD
OB output neuron populations. We optically recorded
GLP-1 neurons: A link between gastrointestinal satiation activity from mitral and deep tufted (MT) cells projecting
signaling and food reward? to piriform cortex and from cholecystokinin-expressing
Diana L. Williams
superficial tufted (ST) cells using Cre-dependent expression
of the genetically-encoded fluorescent calcium indicators
Psychology Department & Program in Neuroscience, Florida GCaMP3 and GCaMP5G. We compared lateral inhibi-
State University Tallahassee, FL, USA tion to these different populations using pairs of monomo-
Central glucagon-like peptide 1 receptor (GLP-1R) stimulation lecular odorants that activated distinct neuron subsets and
suppresses food intake, and hindbrain GLP-1 neurons project their binary odorant mixtures. Both MT and ST popula-
to numerous feeding-relevant brain regions. One such region tions showed detectable mixture suppression in a fraction
is the nucleus accumbens (NAc), which plays a role in reward of neurons (31% for MT vs. 36% for ST cells), reflecting lat-
and motivated behavior. Using immunohistochemical and eral inhibition between cell ensembles with distinct odorant
retrograde tracing techniques in rats, we identified a robust response specificities. However, in these neurons the mag-
projection from GLP-1 neurons in the nucleus of the solitary nitude of this suppression was significantly stronger in MT
tract to the NAc core. Our pharmacologic studies then cells vs. ST cells (70% vs. 25% suppression by the mixture).
provided evidence that exogenous and endogenous GLP-1R Thus, these different OB output populations - which project
stimulation in the NAc core reduces food intake. The NAc is to distinct cortical targets-integrate olfactory information
known to influence food palatability and motivation, while differently. In addition we found that odorants often evoked
GLP-1 neurons have been implicated in mediation of meal- fluorescence decreases in MT somata and glomerular neu-
related gastrointestinal signals. We hypothesize that GLP-1 ropil in a reliable, odorant-specific manner, likely reflecting
released in NAc in response to nutrient ingestion contributes inhibition of spontaneous activity in these neurons. These
to meal-induced satiation, and that GLP-1 action in NAc experiments thus enable us to, for the first time, directly map
reduces the hedonic value of food. To understand the role the spatial organization of lateral inhibition in vivo and to
of endogenously released GLP-1 in the NAc core, we have characterize how this inhibition varies across different cell
been performing detailed behavioral analysis of the effects of types and OB layers as it shapes early odor representations.
blockade of GLP-1R at this site. We find that injection of the Acknowledgements: NIH NIDCD R01 DC006441 NIH
GLP-1R antagonist exendin 9–39 (Ex9) into the NAc core NIDCD 1F32DC012718-01
selectively increases meal size and blunts the satiating effects
of nutrients. In rats consuming sucrose solutions, NAc core #16 PLATFORM PRESENTATIONS: OLFACTION
Ex9 treatment increases initial lick rate and duration of licking
bursts. These measures are directly correlated with palatability, AWnt5 Gradient Patterns the Drosophila Olfactory Map
so this supports the idea that endogenous GLP-1R stimulation Huey Hing1, Yuping Wu2, Lee Fradkin3, Jasprina
in NAc reduces the hedonic value of food. We continue to use Noordermeer3
other behavioral approaches to discern effects on palatability
and motivation for food. Together, our data support the
1
SUNY Brockport/Biology Brockport, NY, USA, 2Stowers
suggestion that the GLP-1 projection to NAc serves as a link Institute for Medical Research Kansas City, KS, USA, 3Leiden
between gastrointestinal satiation signaling and hedonic aspects University Medical Center/Molecular Cell Biology Leiden,
of eating. Acknowledgements: Supported by DK078779 Netherlands
Although the olfactory map facilitates the encoding of odor
#15 PLATFORM PRESENTATIONS: OLFACTION information, how its pattern is specified remains largely
unknown. Recent work has shown that the Drosophila
Lateral inhibition and odor mixture integration among olfactory map is initially formed by spatial segregation of
distinct subpopulations of olfactory bulb output neurons the projection neuron dendrites in the antennal lobe. Here
imaged in vivo we demonstrate that the Wnt5 protein specifies projection
Michael N Economo, Matt Wachowiak
neuron dendrite positions prior to arrival of the olfactory
receptor axons. Wnt5 is expressed by a novel set of guidepost
Brain Institute and Department of Physiology, University of cell neurons which are located at the dorsolateral pole of the
Utah Salt Lake City, UT, USA antennal lobe and generate a dorsolateral to ventromedial
gradient of Wnt5 in the nascent antennal lobe neuropil. Loss #18 PLATFORM PRESENTATIONS: OLFACTION
of wnt5 prevents the appropriate ventral migration of the
Reinforcement of Sexual Attraction through Pheromone-
projection neuron dendrites while over-expression of wnt5
Induced Associative Learning
severely disrupts dendritic patterning. We also show that
Drl, a known Wnt5 receptor, acts cell-autonomously in the Jane L Hurst, Sarah A Roberts, Emma Hoffman, Amanda J
projection neurons to repress the Wnt5 signal. Decreased Davidson, Lynn McLean, Robert J Beynon
drl function causes projection neuron dendrites to inap- University of Liverpool / Institute of Integrative Biology
propriately target ventromedially; a defect which is strongly Liverpool, United Kingdom
suppressed by loss of a copy of the wnt5 gene. We propose
that the Wnt5-secreting guidepost cells act to provide posi- Scents play an integral role in mediating reproductive inter-
tional information to the projection neuron dendrites. Our actions in many species, allowing animals to recognize and
findings demonstrate the importance of a Wnt5 gradient in locate individual conspecifics of the opposite sex, assess their
the patterning of the olfactory map. Acknowledgements: attractiveness and stimulate mating. The urine used by male
DC010916-01 house mice to advertise their location and competitive domi-
nance contains many androgen-dependent volatiles together
with a high concentration of major urinary proteins (MUPs)
#17 PLATFORM PRESENTATIONS: OLFACTION that bind and slow the release of these volatiles. Females that
detect airborne urinary volatiles are attracted to approach
Sexual dimorphism and experience-dependent plasticity in and sniff the scent marks closely, but it is contact with an
mouse vomeronasal neurons atypical male- specific MUP named darcin that reliably elic-
Timothy E Holy, Pei S Xu its female sexual attraction to spend time near male scent.
Darcin will elicit this attraction even in the absence of all
Washington University in St. Louis St. Louis, MO, USA other urinary components, while male urine without darcin
Male and female mice exhibit behaviors particular to is no more attractive than female urine. Importantly, though,
their sex, and these differences presumably reflect sexual darcin acts not only through direct attraction to spend time
dimorphism in neuronal circuitry. However, in terms of near the pheromone itself. It is also highly potent in stimu-
neuronal anatomy or function in the mouse, there are lating associative learning such that animals rapidly learn
relatively few reported differences males and females. Here the same attraction towards the individual airborne odor
we asked whether neurons in the vomeronasal organ, a social detected in association with darcin, targeting attraction to a
odor- and pheromone-sensing neuroepithelium, differed specific male. Darcin also rapidly conditions preference for
functionally between males and females. We performed spatial cues associated with its location, such that mice relo-
exhaustive high-speed calcium imaging from intact cate and prefer to spend time in the site even when scent is
vomeronasal epithelia in 26 imaging volumes from male absent. This conditioned place preference is remembered for
and female mice, measuring the responses of more than a approximately 14 days. Preference for multiple locations can
quarter-million individual sensory neurons. Using a battery be remembered, while the relative amount of darcin in com-
of sulfated steroids, a class of odors originally isolated from peting male scents influences female conditioned preferences.
mouse urine, we found that the large majority of responsive This reveals that pheromone-induced social learning can
neuronal types were found in equal abundance in both males both target and strongly reinforce female sexual attraction
and females. However, we found restricted cases of clear towards individual males even though male mice all produce
sexual dimorphism, including one neuronal type that was the same sex pheromone. Acknowledgements: These studies
more than one-hundred- fold more common in males than were supported by the Biotechnology and Biological Science
in females, by far the strongest dimorphism ever reported in Research Council (BBC503897 and BB/J002631/1) and the
the mammalian central nervous system. We then explored Natural Environment Research Council (NE/G018650), UK
the mechanism generating this dimorphism. Surprisingly,
male/female differences depended entirely on the history of #19 PLATFORM PRESENTATIONS: OLFACTION
sensory experience, as vomeronasal organs from males could
be converted to a pattern indistinguishable from females Behavioral effects of bulbar neuromodulation
simply by prolonged exposure to the odors of female mice. Christiane Linster, Sasha Devore, T Samuel Dillon, Olga
The finding that a strong neuronal dimorphism is determined Escanilla, Matthew Lewis, Laura Manella
entirely by experience, in a sensory system long believed
Cornell University Neurobiology and Behavior Ithaca,
to be devoted to “innate” responses, raises important new
NY, USA
questions about the roles of “nature” and “nurture” in
brain architecture. Acknowledgements: NIH-NIDCD R01 The olfactory bulb (OB) is modulated by a number of
DC005964 NIH-NINDS/NIAAA R01 NS068409 NIH DP1 extrinsic and intrinsic modulatory substances. We here
OD006437 synthesize work from our lab investigating the effects
of cholinergic (ACh), noradrenergic (NE), serotonergic classify participants as normosmic (4–5 correct), hypos-
(5HT), dopaminergic (DA) and hormonal modulation on mic (2–3 correct), or anosmic (0–1 correct). Five years later
olfactory perception. We compare the behavioral effects NSHAP Wave 2 established participant mortality. Logistic
by using well–established behavioral paradigms such as regression used olfactory function at Wave 1 to estimate
olfactory habituation memory and reward-association. the odds of death at Wave 2, controlling for baseline fac-
We find that ACh modulation affects the discrimination of tors known to influence both olfaction and mortality (age,
chemically and perceptually similar odorants (Mandairon gender, education, race/ethnicity, cognitive function, physi-
et al. 2006; Chaudhury et al. 2009), with specific roles cal and mental health, and tobacco/alcohol use). Olfactory
for muscarinic and nicotinic receptors (Devore et al., in identification at Wave 1 strongly predicted subsequent
prep). NE, while also modulating odor discrimination of 5-year all-cause mortality: fully 39% of anosmic subjects
chemically and perceptually similar odorants (Mandairon had died compared to 19% in the hyposmic group and 10%
et al. 2006; see also Doucette et al. 2007), also plays a in the normosmic group (p<0.001). This “dose-dependent”
specific role in regulating signal to noise ratio when very effect was not explained by potential mediating factors,
low concentration odorants are used (Escanilla et al 2010). including number of comorbid diseases, demographics and
In a direct comparison, NE, but not ACh, is shown to cognitive function. In a comprehensive model, anosmic
be functionally important during learning of very low participants had three times the odds of death compared
concentration odorants (Escanilla et al 2012). DA, intrinsic to normosmics (OR 3.00, 95%CI 1.66, 5.42, p<0.001).
to the bulb, modulated odor concentration perception via Moreover, the increased odds of death occurred in all age
activation of D2 but not D1 receptors (Escanilla et al. groups (57–64, 65–74, and 75–85 years). Our results sug-
2009). Blockade of 5HT receptors impaired habituation gest that a simple olfactory test may have clinical utility in
memory formation and specificity, as well as reward-driven predicting 5-year mortality. Possible physiological mecha-
discrimination at low, but not higher odor concentrations nisms linking olfactory loss and death include slowed cellu-
(Lewis and Linster, in prep). Both hormonal (estrodial, lar regeneration and repair or sentinel function as a marker
in mice) and NE manipulations (in rats) extended the of toxic environmental exposures or neurologic decline.
duration of an olfactory habituation memory (Dillon Acknowledgements: The National Institute on Aging
et al, in prep; Manella et al. in prep). In summary, the (R37 AG030481 AG036762 AG029795), the Institute
behavioral effects of OB neuromodulators, while similar for Translational Medicine at The University of Chicago
at first sight, can be dissociated to ascribe specific roles (KL2RR025000), the McHugh Otolaryngology Research
to each of these substances when task demands are Fund, and the American Geriatrics Society.
manipulated in different ways. Acknowledgements: NIH/
NIDCD RO1DC009948 NIH/ NIDCD RO1DC008701
NIH/NIDCD F32DC011974 l′Oreal USA fellowship for #21 PLATFORM PRESENTATIONS: OLFACTION
women in science
The Scent of a Human: A Mosquito Olfactory Receptor
Neuron that Mediates Attraction to Human Skin Odor
#20 PLATFORM PRESENTATIONS: OLFACTION Genevieve M. Tauxe, Anandasankar Ray
Olfactory Dysfunction Predicts 5-year Mortality in Department of Entomology, University of California,

Older Adults Riverside Riverside, CA, USA
Jayant M. Pinto1, Kristen E. Wroblewski2, David W. Kern3, Mosquitoes that feed on humans transmit deadly diseases
L. Philip Schumm2, Martha K. McClintock4 that affect hundreds of millions of people every year. Host-
seeking females use a combination of carbon dioxide and
1
Section of Otolaryngology-Head and Neck Surgery, The
skin odor cues to find human hosts. Even though whole skin
University of Chicago Chicago, IL, USA, 2Department of
odor is attractive by itself, specific compounds have not been
Health Studies, The University of Chicago Chicago, IL,
identified that are attractive in the absence of a CO2 plume.
USA, 3Department of Comparative Human, The University
We use electrophysiology assays to identify a specific olfac-
of Chicago Chicago, IL, USA, 4The Institute for Mind and
tory receptor neuron (ORN) class that responds to whole
Biology, The University of Chicago Chicago, IL, USA
skin odor. From the human odor blend, we identify specific
We sought to determine if loss of olfactory function among compounds that activate this ORN strongly. This response
older US adults is a harbinger of overall health decline is conserved in both Aedes aegypti and Anopheles gam-
predicting 5-year mortality. In 2005–6 The National Social biae, even though these two species, both of major public
Life, Health and Aging Project (NSHAP) interviewed 3,005 health importance, are not closely related. We specifically
adults aged 57–85 (Wave 1) representative of the diverse suppress the activity of this ORN for hours by treatment
aging US population. Olfactory function was measured with a reactive compound that is structurally related to
with 5 Sniffin’ Sticks pens and word/picture prompts to known ligands. After treatment, mosquitoes show deficits
in navigation toward human skin odor in a wind tunnel. #24 SYMPOSIUM: STEM AND PROGENITOR
This is among the first ORN classes shown to play a role CELLS FOR TASTE BUDS — DEVELOPMENT AND
in attraction to skin odor. The receptors expressed in this RENEWAL
ORN are highly conserved, offering hope that compounds
that affect their activity can be used in a new generation of In search of adult taste stem cells
economical and environmentally friendly lures and repel- Karen Yee1, Yan Li1, Kevin Redding1, Ken Iwatsuki2, Robert
lents for mosquito control. Acknowledgements: NIH grant Margolskee1, Peihua Jiang1
RO1AI087785 and R56AI099778. GMT partly supported 1
Monell Chemical Senses Center, 3500 Market Street,
by UC Global Health Institute and Bill and Melinda Gates
Philadelphia, PA, USA, 2Institute for Innovation, Ajinomoto
Foundation.
Co., Inc., Kawasaki-ku, Kawasaki, Japan
Recently, a great deal of progress has been made in identify-
#23 SYMPOSIUM: STEM AND PROGENITOR ing reliable markers for adult stem cells for many regenerative
CELLS FOR TASTE BUDS — DEVELOPMENT AND mammalian tissues. For instance, Lgr5 (leucine-rich repeat-
RENEWAL containing G-protein coupled receptor 5) is a bona fide marker
for adult stem cells in intestine, stomach, and hair follicle. In
Signaling compartments regulate development and the small intestine of mice the Polycomb group protein Bmi1
maintenance of stem, progenitor and differentiated cells in marks another population of stem cells distinct from the
taste papillae and taste buds Lgr5+ stem cells. Taste epithelium also regenerates constantly,
Charlotte Mistretta yet the identity of adult taste stem cells remains elusive. In this
study we set out to determine if Lgr5 and Bmi1 mark adult
Department of Biologic and Materials Sciences, School of taste stem/progenitor cells. We found that Lgr5 is strongly
Dentistry, University of Michigan, Ann Arbor MI expressed in cells at the bottom of trench areas at the base of
Stem and progenitor cell biology is based in spatial and circumvallate and foliate taste papillae and weakly expressed
temporal contexts of compartments or niches where in the basal area of taste buds and that Lgr5-expressing cells
a supporting signaling environment sustains stem and in posterior tongue are a subset of K14-positive epithelial
progenitor properties. Thus for taste bud stem or progenitor cells. Lineage-tracing experiments using an inducible Lgr5-Cre
cells, we must examine: epithelial, mesenchymal and knock-in allele in combination with Rosa26-LacZ and Rosa26-
specialized cells in tongue, papilla and taste bud, in embryo, tdTomato reporter strains showed that Lgr5-expressing cells
early postnatal and adult stages. Principles and knowledge gave rise to taste cells, perigemmal cells, along with self-renew-
about stem/progenitor cell biology in compartment and ing cells at the bottom of trench areas at the base of circumval-
stage specific contexts will be presented. Current thinking is late and foliate papillae. Moreover, using subtype-specific taste
applied with the example of sonic hedgehog (Shh) signaling markers, we found that Lgr5-expressing cell progeny include
in regulating lingual tissue, fungiform papilla and taste bud all three major types of adult taste cells. Our results indicate
development and maintenance, and epithelial/mesenchymal that Lgr5 may mark adult taste stem cells in the posterior
exchanges including contributions from neural crest. The portion of the tongue. In contrast, our lineage tracing experi-
data locate Shh ligand and Shh responding cells; bring ments using Bmi1-Cre; Rosa26-LacZ showed that Bmi1 does
new findings of Shh roles in tongue, papilla and taste bud not mark adult taste stem cells. Acknowledgements: This work
development and maintenance; demonstrate a contribution was supported by NIH grants DC0101842 (P.J.), DK081421
of Shh responding, Gli1 labeled progeny to taste bud cells (R.F.M.), DC003055 (R.F.M.), P30DC011735 (R.F.M) and a
and cells of the papilla; and present distinctive effects of grant from the Commonwealth of Pennsylvania Department
activating Shh transcription factors in adult tongue. Defined of Health (P.J.)
proliferation niches of active Shh signaling suggest that
epithelium and mesenchyme harbor multiple stem and
progenitor cell compartments. In sum, Shh has roles to form #25 SYMPOSIUM: STEM AND PROGENITOR
and maintain fungiform papillae and taste buds, most likely CELLS FOR TASTE BUDS — DEVELOPMENT AND
via stage-specific autocrine and/or paracrine signaling, with RENEWAL
epithelial/mesenchymal interactions. Neural crest cells also
Cell types in adult taste buds: distinct longevities and
contribute to developing and postnatal lingual epithelium and
origins
mesenchyme, including taste papillae and taste buds. Further,
Shh signaling in neural crest cells participates in patterning Nirupa Chaudhari
the tongue. In all, cells originating in several lingual tissue
areas contribute to the stem and progenitor compartments Department of Physiology and Biophysics, and Program
that are active in forming and maintaining taste buds and in Neurosciences, University of Miami Miller School of
papillae. Supported by NIH NIDCD Grant DC000456. Medicine, Miami, FL, USA
Mammalian taste buds are repopulated throughout adult in mammals, recent work in the mouse has shown a latent
life with new cells that are born in the basal epithelium regenerative potential for supporting cells to act as progeni-
immediately surrounding them. Taste buds contain several tors by both cell cycle reentry and transdifferentiation during
molecularly and functionally distinct classes of cells, but the perinatal period, a potential that appears to be rapidly lost
it is unclear whether each class is derived from a separate during early postnatal maturation. In this talk I will present
progenitor cell pool, and has similar longevity. Using high recent findings on the molecular mechanisms that govern cell
resolution confocal microscopy, and 4-color fluorescent cycle reentry of otherwise postmitotic supporting cells, as well
labeling, we examined whether the previously described as mechanisms governing cell fate decisions between sensory
average life-time of taste bud cells applies to each of the hair cells, and the various supporting cell types in the develop-
cell types that make up the taste bud. After a single EdU ing and perinatal organ of Corti. I will also discuss the pos-
injection to label newly born cells, we fitted exponential decay sibilities for future manipulation of supporting cells for the
curves to the disappearance of labeled nuclei from each cell purpose of regeneration of lost sensory hair cells.
type. While sweet-, bitter- and umami-sensing Type 2 cells
displayed a half-life of ≈8 days, sour-sensing neuron-like
Type 3 cells were much longer-lived, with a half-life of over #27 SYMPOSIUM: STEM AND PROGENITOR
22 days. Curiously, many post-mitotic cells had a prolonged
CELLS FOR TASTE BUDS — DEVELOPMENT AND
quiescence inside taste buds before differentiating into mature
RENEWAL
taste cells. We also evaluated whether all taste cell types
arise from a common pool of progenitor cells using lineage-
tracing analyses in adult KERATIN14-cre/ERT;Rosa26- Cellular basis of taste dysfunction following head and neck
YFP mice. Non-taste keratinocytes were produced rapidly irradiation
(within 2 days) from K14+ progenitors; Type I glial-like cells Linda Barlow1,2, Alon Bajayo1,2, Ha Nguyen1,2,3, Mary
became YFP+ ≈10 days after induction, whereas Type II cells Reyland4
contained YFP+ cells only after ≈20 days. In stark contrast, 1
Department of Cell & Developmental Biology, University of
Type III cells did not noticeably acquire the YFP lineage-
Colorado School of Medicine, Aurora CO 80045, USA, 2the
label even 60 days after induction. Thus, while most cells of
Rocky Mountain Taste & Smell Center (RMTSC), University
the taste bud arise from K14+ progenitors, the dynamics of
of Colorado School of Medicine, Aurora CO 80045, USA,
their appearance suggests that several separate progenitor 3
Hanoi Medical University, Department of Histology and
cell pools replenish adult taste buds during normal renewal.
Embryology, No. 1 Ton That Tung, Hanoi, Vietnam, 4Dept
Supported by NIH/NIDCD R01DC6308
of Craniofacial Biology, University of Colorado School of
Dental Medicine, Aurora CO 80045, USA
#26 SYMPOSIUM: STEM AND PROGENITOR Taste dysfunction frequently occurs following radiotherapy
CELLS FOR TASTE BUDS — DEVELOPMENT AND for head and neck cancer. Importantly, patients with reduced
RENEWAL taste function tend to lack appetite and eat far less, leading
to weight loss and a significantly compromised quality of
Development and regeneration of the inner ear: Control of life. To determine the cellular targets contributing to taste
cell division and differentiation of sensory progenitors dysfunction, we developed a mouse model of head and neck
Neil Segil irradiation. We find that proliferating taste bud progenitor
cells are immediate and direct targets of radiation damage.
Division of Cell Biology and Genetics, House Research
Specifically, head and neck irradiation results in: 1) increased
Institute; and Department of Cell and Neurobiology,
apoptosis of progenitors within 24 hours of radiation
University of Southern California.
exposure; and 2) a transient cessation in proliferation, lasting
Loss of the sensory hair cells of the inner ear is the major cause ~3 days. This latter effect results in an interrupted supply
of deafness and balance disorders. Hair cells are extremely of new postmitotic taste cells to buds, and accounts for the
sensitive to various environmental stressors such as ototoxic subsequent reduction in differentiated taste cells seen at 1
drugs noise, and aging. In mammals, the failure to regenerate week post-irradiation. We are now investigating the role of the
these cells is complete, making hair cell loss a major global novel protein kinase C delta isoform (PKCδ) in irradiation-
health problem. In contrast to the failure of regeneration in induced taste epithelial injury. PKCδ is a key regulator of
mammals, non-mammalian vertebrates can efficiently regen- irradiation-induced apoptosis, and suppression of PKCδ
erate sensory hair cells through a combination of direct trans- in vitro and genetic loss in vivo protects salivary gland cells
differentiation of the surrounding supporting cells, as well as from cell death. PKCδ is also implicated in maintenance of
by stimulated cell cycle reentry and subsequent differentiation cell cycle arrest required for DNA repair in UV damaged
of supporting cells into new functional hair cells and sup- human keratinocytes. Thus, we hypothesize that loss of
porting cells. In spite of the failure of hair cell regeneration PKCδ may protect taste progenitor cells from death, and/ or
promote their continued mitosis following irradiation injury. 1

University of Colorado Anschutz Medical Campus,
This model is supported by our pilot data, which suggest Department of Cell & Developmental Biology and the Rocky
that, while PKCδ -/- mice possess normal taste epithelia, in Mountain Taste & Smell Center Aurora, CO, USA, 2University
response to irradiation, the taste bud progenitor population of Pennsylvania School of Medicine, Departments
continues to proliferate. Thus, our studies point to PKCδ of Dermatology and Cell & Developmental Biology
as a potential future target for interventional treatment Philadelphia, PA, USA, 3Institute for Regenerative Medicine
for taste loss in head and neck cancer patients treated with at Scott & White Hospital, Texas A&M University System
radiotherapy. Supported by NIH/ NIDCD R21DC011713 to Health Science Center Temple, TX, USA
LAB and MER, and P30DC004657 to D. Restrepo. In adult mice, taste cells are continually renewed from Keratin
(K)14+ basal keratinocytes. As a population, K14+ basal cells
#28 PLATFORM PRESENTATIONS — POLAK self renew, as well as produce post-mitotic cells which either
YOUNG INVESTIGATOR AWARD WINNERS differentiate into lingual epithelial cells (K13+), or enter buds
and differentiate into type I, II and III taste cells (K8+). We pre-
viously showed that Wnt/β-catenin signaling is active in cells
Trace amine-associated receptors mediate behavioral in and around taste buds of adult mice (Gaillard & Barlow,
aversion in mice 2011), suggesting that this pathway may regulate several
Adam Dewan1, Rodrigo Pacifico1, Dmitry Rinberg2, Thomas aspects of taste cell renewal. To test this idea, we used induc-
Bozza1 ible Cre-lox technology to drive β-catenin gain of function
(GOF) in K14+ basal keratinocytes throughout the tongue
1
Department of Neurobiology, Northwestern University
epithelium, including the fungiform and circumvallate papil-
Evanston, IL, USA, 2Neuroscience Institute, New York
lae (CVP). In the CVP trenches, K13+ cells located between
University Langone Medical Center New York, NY, USA
taste buds vanished in the GOF, and instead all cells within
The Trace Amine-Associated Receptors (TAARs) are a small the CVP epithelium expressed K8. Using immunomarkers
set of evolutionarily conserved main olfactory receptors whose for each of the 3 taste cell types, we found that this expanded
contribution to chemosensory function is not known. Our pre- taste CVP epithelium comprised primarily NTPdase2+ type
vious data show that a majority of the TAARs are mapped I cells, with little or no change in the numbers of type II and
to a discrete group of highly sensitive amine-responsive glo- III cells. Likewise, in the anterior tongue, β-catenin GOF
meruli in the dorsal olfactory bulb of the mouse. Amines have induced multiple K8+ cell clusters within fungiform papillae,
been implicated as social cues and/or predator-derived chem- as well as numerous ectopic K8+ cell clusters in non-taste epi-
osignals in rodents. We have used a combination of behavior thelium; all of these cell clusters were exclusively NTPdase2+,
and in vivo optical imaging to examine the functional conse- and were devoid of expression of markers for type II and III
quences of genetically removing TAAR genes in mice. We find taste cells. Our data indicate that excess Wnt/β-catenin sign-
that deleting all 14 olfactory TAARs abolishes high sensitivity aling drives epithelial progenitors to produce daughter cells
amine responses in the dorsal bulb and eliminates aversion that committed to a taste fate (K8+) at the expense of a non-taste
mice display to structurally diverse amines and to the volatiles fate (K13+), and moreover constrains these newly generated
of predator cat urine. Moreover, removing a single TAAR taste cells to a type I cell fate. We are now examining what
gene (Taar4) produces an odor-specific deficit in sensitivity cellular mechanisms are triggered by excess β-catenin, and
and abolishes behavioral aversion to phenylethylamine—a how these changes in cell renewal result in the GOF pheno-
chemical that is enriched in predator cat urine. Our data reveal type. Acknowledgements: Supported by an American Heart
that the TAARs mediate aversive responses in some behavio- Association fellowship to DG, NIH/NIDCD DC008373 and
ral contexts, and that individual TAAR genes contribute sig- DC012383 to LAB, and DC004657 to D. Restrepo.
nificantly to amine perception in mice. Acknowledgements:
This work was supported by grants from NIH/NIDCD
(R01DC009640), The Whitehall Foundation, and The Brain #30 PLATFORM PRESENTATIONS — POLAK
Research Foundation. YOUNG INVESTIGATOR AWARD WINNERS
Target-defined olfactory bulb output streams isolated using
retrograde infection with recombinant viral vectors
#29 PLATFORM PRESENTATIONS — POLAK
YOUNG INVESTIGATOR AWARD WINNERS Markus Rothermel, Christine Zabawa, Daniela Brunert,
Marta Diaz-Quesada, Matt Wachowiak
Excess Wnt/β-catenin signaling in lingual epithelial Brain Institute and Department of Physiology Salt Lake City,
progenitors drives production of type I taste cells at the UT, USA
expense of all other lingual epithelial cell fates Recombinant viral vectors are an attractive tool for
Dany Gaillard1, Sarah E Millar2, Fei Liu3, Linda A Barlow1 cell-type specific transgene expression, especially when
Cre-dependent vectors are combined with Cre-expressing cell-specific ablation of the G protein Gαo. Female mice
mouse lines. However, cell-type specific promoters do not mutant for Gαo show severe alterations in mate recogni-
always yield sufficient specificity for isolating functionally tion, mating, and reproduction. Male pheromonal cues fail
distinct neuronal populations. In the olfactory system mitral to accelerate puberty onset and estrous synchronization in
and tufted cells (MT) of the olfactory bulb (OB) constitute a these mice. Gαo mutant females exhibit a striking reduction
heterogenous population with distinct dendritic organization, in sexual receptivity or lordosis behavior to males, but gen-
response properties and projections to olfactory cortex. Here, der discrimination seems to be intact. These mice also show
we demonstrate that by combining viral tools with retrograde a loss in scent ownership recognition that requires a learned
infection via their axonal processes, MT cells can be defined association with a nonvolatile ownership signal contained in
by projection target. We injected Cre-dependent AAV the high molecular weight fraction of urine, and they show
vectors into various regions of olfactory cortex in Cdhr1-cre high pregnancy failure rates in the Bruce effect assay. These
mice. Virus injection into anterior piriform cortex (PC) led results indicate that sensory neurons of the Gαo-expressing
to robust and widespread transgene expression in MT cells vomeronasal subsystem, together with the receptors they
throughout the OB. To establish that expression patterns were express and the molecular cues they detect, control a diverse
due to retrograde infection we targeted additional olfactory range of fundamental mating and reproductive behaviors in
cortical areas: injection into posterior PC or posterior female mice. Acknowledgements: This work was supported
cortical amygdala led to expression exclusively in mitral cells by grants from the Deutsche Forschungsgemeinschaft to
with lateral dendrites in the deep external plexiform layer, P.C. (CH 920/2-1), F.Z. (SFB 894) and T.L-Z. (SFB 894),
while injection into medial amygdala led to expression solely the Intramural Research Program of the NIH to L.B.
in mitral cells of the accessory olfactory bulb. Retrograde (Project Z01 ES-101643), and the Volkswagen Foundation
infection was effective using multiple transgenes including (to T.L.-Z.). E.J. was supported by the DFG-funded
GCaMP and ChR2, allowing for optical imaging, optical International Graduate Program GK1326. T.L.-Z. is a
control or optically-assisted electrophysiology of distinct OB Lichtenberg Professor of the Volkswagen Foundation.
output streams defined by their projection target. Retrograde
infection was also effective for projection neurons in other
brain regions. These results establish a valuable, easily-used #32 PLATFORM PRESENTATIONS — POLAK
tool for achieving combinatorial specificity in transgene YOUNG INVESTIGATOR AWARD WINNERS
expression to monitor and manipulate precisely-defined
neuron populations in vivo. Acknowledgements: Supported Congruency matters: dual cortical processing of visual-
by DFG and NIDCD olfactory integration
Kathrin Ohla1,2, Johan N Lundström2,3,4
1
German Institute of Human Nutrition Potsdam-Rehbrücke
#31 PLATFORM PRESENTATIONS — POLAK Nuthetal, Germany, 2Monell Chemical Senses Center
YOUNG INVESTIGATOR AWARD WINNERS Philadelphia, PA, USA, 3Karolinska Institute Stockholm,
Major contribution of Gαo-dependent vomeronasal Sweden, 4University of Pennsylvania Philadelphia, PA, USA
chemoreception to sexual and reproductive behavior in Before ingestion, the visual appearance and odor of a
female mice food constitute its primary sensory inputs. Whether these
Livio Oboti1, Trese Leinders-Zufall1, Eric Jacobi1,3, Lutz distinct sensory events are perceived as one entity shapes
Birnbaumer2, Frank Zufall1, Pablo Chamero1 their impact on subsequent food choice and possibly
also food perception. We hypothesized that the degree of
1
Department of Physiology, University of Saarland School of
perceived concordance of the sensory inputs influences
Medicine Homburg, Germany, 2Laboratory of Neurobiology,
multisensory integration processes. To date, the behavioral
Division of Intramural Research, National Institutes of
consequences and brain mechanisms underlying these
Health Research Triangle Park, NC, USA, 3Deutsches Zentrum
processes are poorly understood and were investigated
für Neurodegenerative Erkrankungen (DZNE) Heidelberg,
with the present study. We used electrical neuroimaging
Germany
analyses of the electroencephalographic (EEG) responses
Optimal reproductive fitness is essential for the biologi- following olfactory-visual stimulation in humans. Stimuli
cal success and survival of species. The vomeronasal organ were odor-image pairs presented as 100% congruent, 50%
(VNO) is strongly implicated in the display of sexual and congruent and 100% incongruent. Participants rated the
reproductive behaviors in female mice, yet the role that apical stimuli for congruence, pleasantness and intensity. As
and basal vomeronasal neuron populations play in control- expected, concordant stimulus pairs were perceived as
ling these gender-specific behaviors remain largely unclear. more congruent and as more pleasant than mixed and
To dissect neural pathways underlying these functions, we incongruent pairs. Waveform analysis yielded significant
genetically inactivated the basal VNO layer using conditional, amplitude augmentation for congruent as compared to
incongruent odor-image pairs between 120–200ms post point to mPFC as a cortical area involved in coding of palat-
stimulus onset. Source analysis revealed that the activation ability. Acknowledgements: NIDCD Grant R01-DC010389
differences origin in visual cortex and left inferior temporal
gyrus. Later differences were observed between 400–700ms
in the parietal lobe, lateral frontal cortex and the bilateral #34 SYMPOSIUM: NEW APPROACHES TO
insula. Concordance between olfactory-visual stimuli was PHYSIOLOGY AND BEHAVIOR IN AWAKE
associated with increased pleasantness and activation in RODENTS
unimodal visual and olfactory areas as well as in multimodal
areas. The results suggest that cross-modal integration is New approaches to physiology and behavior in awake
a dynamic process regulated by both unisensory as well rodents
as higher order integration areas and that this process is
Stephen Shea1, Dinu Albeanu1, Alfredo Fontanini2,
modulated by learned associations and perceived congruence
Venkatesh Murthy3, Andreas Schaefer4, Ben Strowbridge5
between the sensory inputs.
1
Cold Spring Harbor Laboratory Cold Spring Harbor,
NY, USA, 2Stony Brook University Stony Brook, NY, USA,
#33 PLATFORM PRESENTATIONS — POLAK 3
Harvard University Cambridge, MA, USA, 4MPI Heidelberg
YOUNG INVESTIGATOR AWARD WINNERS Heidelberg, Germany, 5Case Western Reserve University
Cleveland, OH, USA
Processing of hedonic and chemosensory features of taste
in medial prefrontal and gustatory cortices As our understanding of neural circuitry grows more sophisti-
cated, there is increasing interest in studying neuronal process-
Ahmad Jezzini, Luca Mazzucato, Giancarlo La Camera, ing during volitional behavior in awake animals. At the same
Alfredo Fontanini time, recent years have seen the emergence of many new tech-
Dept of Neurobiology & Behavior, SUNY Stony Brook, niques that allow unprecedented access to observe and control
NY, USA neural activity with spatiotemporal precision. These sensitive
techniques can be difficult to implement and chemical stimu-
The gustatory cortex (GC) is the main cortical recipient of
lus control can be problematic in freely behaving animals. For
taste-related signals, however it is not the only cortical area
these reasons, many labs are developing hybrid approaches
involved in processing taste. Upon elaborating gustatory
in head-fixed preparations that fuse these new tools with rich
signals, GC sends information to the orbitofrontal cortex
behavioral paradigms in awake, behaving animals. The goal of
(OFC) and the medial prefrontal cortex (mPFC). While con-
this symposium will be to bring together investigators who are
siderable amount of work has been devoted to understanding
applying high resolution tools in awake animals to break new
how GC and OFC process different features of a gustatory
ground in our appreciation of the state modulation of chem-
experience, less is known regarding the role of mPFC. We
osensory circuits and their governance of behavior.
investigated the involvement of mPFC in taste processing
by comparing its responses to gustatory stimuli with those
observed in GC. Eight rats were chronically implanted with
movable bundles of electrodes in the ipsilateral mPFC and #35 SYMPOSIUM: NEW APPROACHES TO
GC. Extracellular recordings were performed while 4 tastes PHYSIOLOGY AND BEHAVIOR IN AWAKE
were passively delivered through intraoral cannulae. The RODENTS
results showed significant taste related activity in mPFC. In
comparison to GC, mPFC was less responsive to taste (49% Dramatic state-dependency of the activity of granule cells
of mPFC units responded to taste vs 75% in GC) and fir- in the mouse main olfactory bulb
ing rates were lower in mPFC than GC. While taste selec- Stephen D Shea, Brittany N Cazakoff, Kerensa L Crump,
tivity was more pronounced in GC than mPFC, activity in
mPFC appeared more strongly modulated by palatability. Billy Y Lau Cold Spring Harbor Laboratory Cold Spring
Results from a classification analysis revealed that units in Harbor, NY, USA
GC decode taste quality more successfully than mPFC, and It is becoming increasingly clear that olfactory represen
mPFC units encode tastes according to their palatability. tations in the main olfactory bulb (MOB) are substantially
Analysis of the time course of responses further revealed reformatted in awake rodents. In addition to elevated rates and
that, contrary to the GC where palatability coding occurs altered temporal structure in the spike discharge of mitral/
within the first 2 seconds following taste delivery, palatability tufted cells (MT), the wakeful state is marked by a fusion
coding in mPFC lasts for as long as 5 seconds. Furthermore, of sensory input-driven responses with activity that reflects
analysis of firing rates evoked by tastes revealed a bias toward attention, experience, and behavioral task contingencies. It
aversive stimuli in mPFC. Altogether our results suggest a seems likely that as a key target of many neuromodulatory
role of the mPFC in taste processing and more specifically systems and corticofugal feedback pathways, the extensive
network of inhibitory MOB granule cells (GC) is instr and odor specific, GCs showed both broad, and narrowly
umental in the state-dependent sculpting of MT activity tuned inhibitory responses. Further, a significant fraction
patterns. Despite this predicted critical role, few published (~25%) of GCs exhibited excitatory OFF responses, inde-
studies have demonstrably made electrophysiological pendent of stimulus duration. We are currently investigat-
recordings from these small cells. Moreover, none have ing how response properties of GCs and feedback fibers are
been reported in awake animals. As a first step towards shaped by odor experience and during reinforcement learn-
closing this gap, we recently developed reliable methods for ing. Additionally, we are employing pharmacological and
recording and labeling GC in awake, head-fixed mice. Our optogenetic approaches to modulate the feedback fibers,
preparation allows us to directly compare the activity and while simultaneously monitoring granule cell activity.
sensory responses of the same GC during wakefulness and
inhalant anesthesia. Our data reveal that GC in awake mice #37 SYMPOSIUM: NEW APPROACHES TO
are dramatically more spontaneously active, and exhibit
PHYSIOLOGY AND BEHAVIOR IN AWAKE
stronger, more broadly-tuned sensory responses that include
both increases and decreases in spike rate. Under either of RODENTS
two pharmacologically-distinct anesthetics, many of these Odor-guided behaviors in head-restrained and
cells emit very few spontaneous or stimulus-driven spikes. freely-moving mice
Those that have somewhat higher spontaneous rates still Venkatesh N Murthy1,2, Dan Rokni1,2, David H. Gire1,2, Daniel
exhibit little response to odors. We are currently quantifying Millman1,2
the variable respiratory coupling of GC in awake animals,
and assessing the effects of stimulus novelty or familiarity
1
Harvard University, Molecular & Cellular Biology
on GC odor responses. Cambridge, MA, USA, 2Harvard University, Center for Brain
Science Cambridge, MA, USA
Olfaction plays a central role in guiding behavior in many
#36 SYMPOSIUM: NEW APPROACHES TO animals, including the common laboratory mammalian
PHYSIOLOGY AND BEHAVIOR IN AWAKE models – rats and mice. There is a renewed excitement about
RODENTS studying the neural basis of such behaviors, which entails
developing behavioral tasks under controlled conditions
Response properties of cortico-bulbar feedback and granule where neural activity can be recorded and manipulated. We
cells in awake head-fixed mice have trained mice to perform odor tasks while their heads
Dinu F Albeanu, Hong goo Chae, Gonzalo H Otazu are restrained, which allows stable electrophysiological
recordings, high-resolution optical imaging and optogenetic
Cold Spring Harbor Laboratory Cold Spring Harbor, NY, USA
manipulation. In one such task, head-restrained mice
Sensory circuits integrate inputs from the environment, as can be trained to recognize target odorants embedded in
well as feedback signals from higher brain regions, in a close unpredictable and variable background mixtures. We have
loop manner. The interplay of feed-forward and feedback also developed strategies to study odor-guided behaviors in
signals has been proposed to be fundamental for learning freely moving animals, including spatial navigation. We will
and memory recall. Though rich cortical feedback projec- present a detailed analysis of these behaviors in our talk, and
tions innervate the mouse olfactory bulb, to date, little is some initial efforts in recording and imaging neural activity
known about their contribution to olfactory processing. under these conditions. Acknowledgements: R01DC011291
Cortico-bulbar feedback primarily targets the granule cells
(GC), which form extensive dendro-dendritic synapses with #38 SYMPOSIUM: NEW APPROACHES TO
the mitral/tufted cells. To examine how cortical feedback
PHYSIOLOGY AND BEHAVIOR IN AWAKE
shapes processing in the bulb, we used genetically encoded
calcium indicators (GCaMP3&5) and multiphoton imaging RODENTS
to monitor the odor evoked responses of feedback fibers Baseline states and odour evoked responses of mitral and
and granule cells, in awake head-fixed mice. GCs and feed- tufted cells in the awake mouse
back fibers showed rich spontaneous activity and diverse Andreas T. Schaefer1,2,3, Anja Schmaltz1,2, Izumi Fukunaga1,
excitatory and inhibitory odor responses. On average, to our Mostafa Abdelhamid1,2, Mihaly Kollo1
stimulus panel (up to 30 odors), we observed 54% purely
excitatory and only 17% purely inhibitory responses in
1
Behavioural Neurophysiology, MPI for medical research
the GCs, while the two response types were equally com- Heidelberg, Germany, 2Dept Anatomy & Cell Biology,
mon in the feedback fibers. Interestingly, both GCs and University Heidelberg Heidelberg, Germany, 3Division of
feedback fibers, that showed spontaneous activity, were Neurophysiology, MRC-NIMR London, United Kingdom
inhibited upon odor presentation, irrespective of stimu- Odour stimuli evoke activity patterns in the olfactory bulb,
lus identity. While inhibition of feedback fibers was sparse which are transformed in multiple steps by local microcircuits,
before arriving to the cortex. Since inhibitory and excita- conditioning can result in robust anticipatory responses in the
tory synapses both exhibit short-term plasticity, the baseline GC of restrained as well as freely moving animals. Experiments
activities and properties of odour-evoked responses strongly investigating the role of thalamic and limbic inputs in
determine, how much individual neurons can influence infor- generating cue-responses will be presented. After a discussion
mation processing. Therefore, gaining an unbiased picture of the system-level underpinnings for the integration of taste-
about the states of different neurons in the behaving animal coding and general expectation, I will show novel evidence that
is essential for understanding input transformation. We have neurons in GC can also learn to encode specific expectation.
performed blind whole-cell patch-clamp recordings in awake Results from neural recordings in rats performing a two-cues
head-fixed mice to gain detailed and unbiased measurements auditory go/no-go task will be discussed. I will present data
of spontaneous and odour-evoked activity. Mitral and tufted showing that cue responses are outcome specific and decrease
cells (M & TCs) show great diversity in their baseline rest- significantly after partial extinction of cue-taste contingencies.
ing membrane potentials and spike rates. Both in awake and Taste responses will be analyzed in neurons with different
anesthetized animals, a large proportion (>33%) of cells have profiles of cue responsiveness and a relationship between
very low spontaneous firing rates (<1 Hz). In awake animals, responses to sucrose-anticipating cues and to sucrose itself
M&TCs exhibit both inhibitory and excitatory subthreshold will be established. Altogether our data will further establish
and suprathreshold odour-evoked responses. Strong, phasic the function of GC in integrating sensory and anticipatory
excitatory responses can be observed in a subset of cells with signals and will provide a first analysis of the system-level
hyperpolarized membrane potentials. TCs exhibit strong mechanisms mediating this function. Acknowledgements:
excitatory responses more frequently than MCs. Granule cells NIDCD R01-DC010389
are characterized by exceedingly hyperpolarized resting mem-
brane potentials (−66.3 ± 3.4 mV), and the virtually complete
lack of spontaneous spikes. In response to specific odours they #40 SYMPOSIUM: NEW APPROACHES TO
show prolonged subthreshold and only rarely suprathreshold PHYSIOLOGY AND BEHAVIOR IN AWAKE
excitatory responses. This large heterogeneity in base firing RODENTS
rates and odour-evoked responses suggests that different sub-
populations of principal neurons, defined by their internal Differential synaptic control of mitral and tufted cell output
states, have distinct roles in local processing and points to the pathways in the olfactory bulb
importance of intracellular recording techniques for study- Ben W. Strowbridge
ing olfactory networks. Acknowledgements: Max-Planck-
Department of Neurosciences, Case Western Reserve
Society, Humboldt Foundation, DFG ExcellenzCluster
University School of Medicine Cleveland, OH, USA
CellNetworks, Bauer Foundation, Gottschalk Foundation,
BMBF, DFG SPP 1392 Neurons in the gustatory cortex (GC) do more than just
encoding chemosensory signals coming from the thalamus.
Evidence from behavioral and electrophysiological experi-
#39 SYMPOSIUM: NEW APPROACHES TO ments indicates that GC neurons can also process affective
PHYSIOLOGY AND BEHAVIOR IN AWAKE information from limbic regions. The integration between
RODENTS sensory and limbic afferents is believed to explain why neu-
ral responses in GC of alert animals are plastic, multimodal
Integration of anticipatory and gustatory signals in the and rich of reward-related information. My presentation will
gustatory cortex of alert rats begin by reviewing evidence that activity in GC can be strongly
Alfredo Fontanini affected by expectation and by auditory cues anticipating the
general availability of taste. Data from different behavioral
SUNY Stony Brook Stony Brook, NY, USA
paradigms will be used to demonstrate that both classical and
Neurons in the gustatory cortex (GC) do more than just instrumental conditioning can result in robust anticipatory
encoding chemosensory signals coming from the thalamus. responses in the GC of restrained as well as freely moving ani-
Evidence from behavioral and electrophysiological experiments mals. Experiments investigating the role of thalamic and lim-
indicates that GC neurons can also process affective information bic inputs in generating cue-responses will be presented. After
from limbic regions. The integration between sensory and a discussion of the system-level underpinnings for the integra-
limbic afferents is believed to explain why neural responses tion of taste-coding and general expectation, I will show novel
in GC of alert animals are plastic, multimodal and rich of evidence that neurons in GC can also learn to encode specific
reward-related information. My presentation will begin by expectation. Results from neural recordings in rats perform-
reviewing evidence that activity in GC can be strongly affected ing a two-cues auditory go/no-go task will be discussed.
by expectation and by auditory cues anticipating the general I will present data showing that cue responses are outcome
availability of taste. Data from different behavioral paradigms specific and decrease significantly after partial extinction of
will be used to demonstrate that both classical and instrumental cue-taste contingencies. Taste responses will be analyzed in
neurons with different profiles of cue responsiveness and a development. However, whether BDNF has any function
relationship between responses to sucrose-anticipating cues in the adult gustatory system or influences degeneration
and to sucrose itself will be established. Altogether our data and regeneration after nerve injury is unclear. To address
will further establish the function of GC in integrating sen- these issues, we inducibly removed BDNF in adulthood.
sory and anticipatory signals and will provide a first analy- In experimental animals, Bdnf expression decreased to 4%
sis of the system-level mechanisms mediating this function. of control mice in the lingual epithelium and geniculate
Acknowledgements: NIDCD R01-DC010389 ganglion (p<0.001). We found no effect on taste bud
morphology at 30 days following BDNF removal. However,
#41 PLATFORM PRESENTATIONS: TASTE 70 days following BDNF removal, P2X3-positive gustatory
innervation to individual taste buds was reduced by nearly
Characterization of Testicular Bitter Taste Receptor- half (p<0.001) and both taste bud volume (p<0.05) and taste
Mediated Signal Transduction cell number decreased 30% (p<0.01). We unilaterally cut the
Jiang Xu, Liquan Huang chorda tympani nerve (CTX) two weeks after BDNF removal
to determine whether degeneration and/ or regeneration
Monell Chemical Senses Center Philadelphia, PA, USA
requires BDNF in adulthood. Two weeks after CTX, all
Bitter taste receptors were initially isolated from mammalian P2X3-positive and most Tuj1-postive nerve innervation to
taste bud cells in the oral cavity and are believed to function as the taste bud was gone (p<0.001) and taste bud number and
a gatekeeper to prevent poisonous substances from ingestion. size were decreased (P<0.05). There was no effect of BDNF
In addition to taste buds, however, these receptors have also removal on the degree of gustatory degeneration. However,
been found in some extraoral tissues such as the respiratory preliminary data suggests that 60 days following CTX, much
epithelium and gastrointestinal tract. We detected the expres- of the P2X3-positive and Tuj1-postive nerve innervation
sion of these receptors in both human and murine testis, and of control mice has returned and both taste bud number
found that male germ cells including spermatids and epididy- and taste bud size had increased by about 40% compared
mal sperm can respond to bitter tasting substances by increas- to two weeks post-CTX. However, mice lacking BDNF had
ing intracellular calcium concentrations in a dose-dependent no P2X3 labeled nerve fibers and few TuJ1-positvie nerve
manner, and these calcium responses can be blocked by spe- fibers returning to fungiform papillae. In addition, both
cific bitter blockers or the ablation of the G protein gustducin. taste bud number and taste bud size remained the same as
Further characterization of these responses with pharmaco- two-weeks following CTX. These experiments demonstrate
logical agents indicated that depletion of intracellular calcium that BDNF is required for maintenance of taste innervation,
stores with thapsigargin nearly abolished the calcium responses and regeneration of gustatory nerve fibers in adulthood.
to the bitter compounds tested whereas the absence of the Acknowledgements: NIH DC006938
extracellular calcium did not affect the response amplitudes.
Pretreatment of the cells with an adenylate cyclase inhibitor,
MDL12,330A, also did not alter the response to caffeine. Thus #43 PLATFORM PRESENTATIONS: TASTE
our data suggested that testicular bitter taste receptors employ
the G protein gustducin and calcium channels in the endoplas- Regulation of sugar habits by dorsolateral striatal circuits
mic reticulum to mediate calcium responses to bitter tasting Sara Hernandez1, Luis Tellez1,2, Wenfei Han1,2, Ivan De
compounds. Further studies are in progress to elucidate the Araujo1,2
possible roles of these receptors in the reproductive system.
The J.B. Pierce Laboratory New Haven, CT, USA,
1
Acknowledgements: This work was supported by National
Department of Psychiatry Yale University School of
2
Institutes of Health Grant R01 DC007487 to L.H., by NIH-
Medicine New Haven, CT, USA
NIDCD Core Grant P30 DC011735 to R. Margolskee in
support of Monell Core Facilities, and by National Science Despite the introduction of low-calorie sweeteners to the
Foundation Equipment Grant DBI-0216310 to N. Rawson in market, sugar consumption remains a major factor driv-
support of Monell’s Confocal Microscopy. ing obesity rates throughout industrialized and emerging
economies. Experimental psychologists named “habits”
#42 PLATFORM PRESENTATIONS: TASTE those inflexible behaviors, such as sugar intake, that per-
sist despite their harmful consequences. The acquisition
BDNF Maintains Adult Taste Innervation and Is Required For and expression of behavioral habits depend on the integ-
Taste Nerve Regeneration After Injury rity of dopaminergic signaling in the lateral aspect of the
Lingbin Meng, Robin Krimm dorsal striatum. Specifically, dopamine release in dorsolat-
eral striatum is required for behavioral patterns to become
University of louisville,ASNB department Louisville, KY, USA
insensitive to devaluation. We are investigating the role of
Brain derived neurotropic factor (BDNF) is required for dorsolateral striatal circuits in regulating intake of artificial
the gustatory neuron survival and target innervation during sweeteners and sugars. We first determined whether animals
exposed to daily access to sugar or artificial sweeteners #45 PLATFORM PRESENTATIONS: TASTE

become resistant to reward devaluation, i.e. develop habit-
like intake patterns. Adult mice were given 1h ad libitum Sonic Hedgehog Drives Nerve-Independent Formation of
access to sucralose paired to intra-gastric infusions of either Adult Taste Buds
glucose or sucralose. After a 14-days conditioning period, David Castillo1,2, Kerstin Seidel3, Ernesto Salcedo1, Christina
animals were tested after reward devaluation produced by Ahn4, Frederic J. de Sauvage4, Ophir Klein3,5,6, Linda Barlow1,2
a glucose intra-gastric preload. Mice challenged with glu- 1
Dept of Cell and Developmental Biology and the Rocky
cose, but not with sucralose, maintained their daily intake
Mountain Taste and Smell Center, University of Colorado
after the preload, suggesting that sugar but not artificial
School of Medicine Aurora, CO, USA, 2Graduate Program
sweeteners favor the formation of habits. Accordingly, intra-
in Cell Biology, Stem Cells and Development, University
gastric glucose but not sucralose produced robust dopamine
of Colorado School of Medicine Aurora, CO, USA,
release in dorsolateral striatum, suggesting that sugar calo- 3
Program in Craniofacial and Mesenchymal Biology and
ries but not sweetness alone control dopamine efflux in this
Department of Orofacial Sciences, University of California,
circuit. Current experiments involve evaluating whether
San Francisco San Francisco, CA, USA, 4Dept. of Molecular
stimulating dopamine signaling in dorsolateral striatum is
Biology, Genentech Inc South San Francisco, CA, USA,
sufficient for the formation of artificial sweetener habitual 5
Department of Pediatrics, University of California, San
intake. Our data indicate that the dorsolateral striatum is
Francisco San Francisco, CA, USA, 6Institute for Human
critical for the inflexible nature associated with sugar intake.
Genetics, University of California San Francisco San
Acknowledgements: NIDCD grant DC009997
Francisco, CA, USA
Sonic hedgehog (Shh) is a key regulator of cell prolifera-
#44 PLATFORM PRESENTATIONS: TASTE tion and differentiation. In the developing tongue, Shh
patterns embryonic taste buds, but its role in mainte-
CALHM1 ion channel mediates purinergic nance of adult taste buds is unknown. We used an induc-
neurotransmission of sweet, bitter and umami tastes ible, Cre-lox system, where K14CreER, when induced
J. Kevin Foskett1, Akiyuki Taruno1, Zhongming Ma1, Ichiro by tamoxifen, drives mosaic expression of Shh in taste
Matsumoto2, Michael G. Tordoff2, Philippe Marambaud3 and lingual basal keratinocytes. Unexpectedly, ectopic
Shh expression induced ectopic taste bud formation in
1
Dept Physiology, University of Pennsylvania Philadelphia,
the lingual epithelium, a condition never encountered in
PA, USA, 2Monell Chemical Sciences Center Philadelphia,
wild type mice. Tongues examined for a general marker
PA, USA, 3The Feinstein Institute for Medical Research
of taste cells, cytokeratin (K)8 at 1–4 weeks post- induc-
Manhasset, NY, USA
tion, had numerous K8+ cell clusters found external to
Recognition of sweet, bitter and umami tastes requires the fungiform papillae. Further, using markers of 3 differen-
non- vesicular release from taste bud cells of adenosine tiated taste cell types, we found type I, II and III taste
5′-triphosphate (ATP), which acts as a neurotransmitter to cells in these ectopic cell clusters, revealing that these
activate afferent neural gustatory pathways. However, how clusters were indeed taste buds. As taste bud differentia-
ATP is released to fulfill this function is not fully understood. tion and maintenance are nerve-dependent, we assessed
Here we show that calcium homeostasis modulator 1 if formation of ectopic buds was likewise nerve-depend-
(CALHM1), a voltage-gated ion channel, is indispensable ent. Using P2X2, a marker of gustatory innervation,
for taste stimuli-evoked ATP release from sweet-, bitter- abundant neurites were present in fungiform buds, but
and umami-sensing taste bud cells. Calhm1 knockout mice were lacking from ectopic buds. However, using PGP9.5,
have severely impaired perceptions of sweet, bitter and a general marker of lingual nerve fibers, neurites were
umami compounds, whereas sour and salty taste recognition found in the vicinity of ectopic buds, although many
remains mostly normal. Calhm1 deficiency affects taste ectopic buds were devoid of innervation. Therefore, we
perception without interfering with taste cell development or developed an unbiased, automated MATLAB script to
integrity. CALHM1 is expressed specifically in sweet/bitter/ quantify PGP9.5+ neurites in and around K8+ taste
umami-sensing type II taste bud cells. Its heterologous buds. We found no relationship between innervation
expression induces a novel ATP permeability that releases and the presence of ectopic taste buds, controverting the
ATP from cells in response to manipulations that activate general consensus that innervation is required for adult
the CALHM1 ion channel. Knockout of Calhm1 strongly taste bud differentiation. Moreover, we demonstrate
reduces voltage-gated currents in type II cells and taste- that Shh supplied via local epithelial cells is sufficient to
evoked ATP release from taste buds without affecting the drive differentiation of the entire taste bud cell comple-
excitability of taste cells to taste stimuli. Thus, CALHM1 ment in ectopic locations in a nerve-independent man-
is a voltage-gated ATP release channel required for sweet, ner. Acknowledgements: NIH/NIDCD DC008373 and
bitter and umami taste perception. DC012383 to LAB, and DC004657 to D. Restrepo
#46 PLATFORM PRESENTATIONS: TASTE a longitudinal analysis to determine the effects of 40% CR

on rat taste bud morphology and expression of sweet taste
Phenotypic Characterization of a Caudal to Rostral
modulators. Immunohistochemical analyses of the effects of
Intrasolitary Pathway
40% CR on taste buds were made with 5-, 17- and 30-month
Joseph M Breza, Zhixiong Chen, Joseph B Travers, Susan P old male Fisher 344 rats. No significant effects of 40% CR
Travers on taste bud size and number of taste cells per taste bud were
The Ohio State University Columbus, OH, USA noted. However, 30-month old rats (both ad libitum (AL)
and calorie restriction (CR) groups) possessed smaller taste
Interactions between gustatory and visceral signals bud size and fewer taste cells per bud than 5- (significant) or
modulate ingestive behavior. The current study explored 17- (non-significant) month old CR or AL rats. There was no
such interactions in the nucleus of the solitary tract (NST) significant effect of 40% CR or aging on Type 1 (NTPDase 2),
where gustatory and visceral primary afferents terminate in Type 2 (PLC-beta 2), or Type 3 (NCAM) taste cells marker
largely separate rostral (rNST) and caudal (cNST) regions, expression. In contrast, both 40% CR and AL 30-month old
respectively. Previous anterograde tracing suggests an rats demonstrated significantly lower Type 4 (Shh) taste cell
intrasolitary projection from cNST to rNST (Karimnamazi marker expression. We found that α-gustducin expression
et al.,’98) but the phenotypes of these connections have was significantly higher in 5-month old 40% CR rats com-
not been characterized. We made electrophysiologically– pared to AL, with similar trends for T1r3 and glucagon- like
guided iontophoretic injections of the retrograde tracer, peptide 1 (GLP-1) expression. However, T1r3, GLP-1 and
Fluoro-Gold (FG), into gustatory NST in wild- type and α-gustducin expression were decreased in 30-month old 40%
transgenic mice expressing EGFP under the control of the CR rats compared to age-matched AL rats. Leptin recep-
GAD67 promoter. Sections were immunostained for FG tor expression was significantly higher in 17- and 30-month
and tyrosine hydroxylase (TH) to identify catecholaminergic old 40% CR rats, compared to age-matched AL rats. Our
neurons or PHOX2b, a transcription factor that colocalizes findings suggest that short- and long-term CR elicit differen-
with a subset of glutamatergic cells. Retrogradely–labeled tial responses on rat taste bud morphology and sweet taste
cNST neurons extended caudal to obex. At a mid– modulator expression. This is likely due to long- and short-
postremal level, where several neuron types, including A2 term calorie intake and metabolic homeostatic adaptations
catecholaminergic cells important in satiety are prominent, to the CR regimen. Acknowledgements: This work was sup-
the intrasolitary pathway contained both PHOX2b/ ported entirely by the Intramural Research Program of the
excitatory (~65 ± 2.7%) and GAD67/ inhibitory (27 ± 2.5%) National Institute on Aging, National Institutes of Health.
projections, with PHOX2b neurons located preferentially in
the medial and intermediate NST and the GAD67 neurons
distributed more laterally, including the ventral lateral NST.
#48 SYMPOSIUM: EXPERIENCE DRIVEN
A smaller population (7 ± 1.2%), presumably a subset of the
PHOX2b cells, was double-labeled for FG and TH. Parallel
PLASTICITY OF THE OLFACTORY SYSTEM
experiments in rats showed a similar distribution of FG Experience Driven Plasticity of the Olfactory System
neurons labeled with dopamine beta hydroxylase implying Xavier Grosmaitre
that most TH–labeled neurons in mice are noradrenergic.
In addition, in vitro recordings from rNST demonstrate that CSGA, UMR 6265 CNRS - 1324 INRA - Université de
norepinephrine has a suppressive effect on solitary tract– Bourgogne Dijon, France
evoked responses, suggesting a functional role for the A2 The olfactory system has been thoroughly investigated
projection. Acknowledgements: Supported by DC00416 & during several decades. A number of its functions have
T32DE014320 been well described such as i) the olfactory transduction
pathways; ii) the central odor processing and cerebral and
#47 PLATFORM PRESENTATIONS: TASTE neural networks; iii) the mechanisms of neurogenesis and
synaptic plasticity. The olfactory system reveals itself as a
Longitudinal analysis of 40% calorie restriction on rat taste flexible sensory processing system. While the implication
bud morphology and expression of sweet taste modulators of experience and internal state on olfactory detection and
Huan Cai, Wei-na Cong, Rui Wang, Caitlin Daimon, Patrick processing has been explored, some points remain unclear:
Chirdon, Rafael deCabo, Stuart Maudsley, Bronwen Martin whether experience can modulate specific populations
of olfactory sensory neurons (OSNs) and olfactory bulb
National Institute on Aging Baltimore, MD, USA
(OB) input and output maps. New tools were recently
Taste perception is strongly associated with body weight developed to analyze the plasticity of the olfactory system
and metabolic state. Caloric restriction (CR) is a well- such as genetic labeling of specific population of OSNs
characterized intervention that reduces body weight and (i.e. expressing specific OR), molecular biology tools (qRT-
improves metabolic function. In this study, we performed PCR), genetic labeling for synaptic transmission imaging
and in vivo imaging. This symposium focuses on the dynamic plasticity in individual MOR23 neurons. Taken together,
nature of olfactory processing: the effects of experience on our data suggest that the olfactory epithelium presents
how odors are processed from olfactory sensory neurons to deep anatomical, molecular and functional changes when
higher-order structures and how recent techniques are used chronically exposed to odorant molecules in early stage of
to investigate these questions in different structures and life. Acknowledgements: Funding was provided by CNRS
models. In mice, we will discuss: i) the effects of olfactory (ATIP grant), by Conseil Régional de Bourgogne (FABER
experience on specific OSNs populations and synaptic and PARI grants), and by Université de Bourgogne (BQR
transmission; ii) the effects of emotional experience on program).
OSNs function; ii) the importance of the balance between
activity, regeneration and remodeling for OB plasticity; iii)
the use of long-term imaging of odor representations in #50 SYMPOSIUM: EXPERIENCE DRIVEN
awake mice to explore olfactory plasticity. Finally, plasticity PLASTICITY OF THE OLFACTORY SYSTEM
induced by olfactory experience on behavior and brain in an
Insect model will be presented. We aim to present the state Altered olfactory sensory neuron physiology following
of the art about the olfactory system as a flexible, adaptable odor exposure or olfactory fear conditioning in vivo
and plastic system. John P. McGann, Michelle C. Rosenthal, Marley D. Kass,
Andrew H. Moberly
Rutgers University / Psychology Department Piscataway,
#49 SYMPOSIUM:EXPERIENCE DRIVEN NJ, USA
Experience-dependent plasticity is increasingly understood
Postnatal Odorant Exposure Induces Peripheral Olfactory to occur throughout adulthood in mammalian sensory
Plasticity systems. This talk will present data from experiments that
Xavier Grosmaitre used optical imaging to longitudinally assess the effects of
odorant exposure and emotional learning on the physiology
CSGA, UMR 6265 CNRS - 1324 INRA - Université de
of olfactory sensory neurons (OSNs) in vivo in individual
Bourgogne Dijon, France
adult mice. In 1 experiment, mice expressing the fluorescent
Olfactory sensory neurons (OSNs) form an interface exocytosis indicator synaptopHluorin in mature OSNs
between the environment and the brain, converting chemi- underwent a baseline imaging session in which a chronic
cal information (odorants) into electrical signals and send- cranial window was implanted in the skull overlying the
ing these signals to the brain. Little is known about the olfactory bulbs and the OSN synaptic output into olfactory
consequences of long term odorant exposure on OSNs. Our bulb glomeruli was visualized during the presentation of
goal is to understand the anatomical, molecular and physi- a panel of 4 odorants (2 esters, 1 aldehyde, and 1 ketone).
ological effects of odorant exposure at the cellular level and Mice then spent a week in either an odorant-exposure
more precisely in the context of an early postnatal olfac- chamber in which 1 of the esters was presented repeatedly
tory exposure. We focus our work on specific populations with a 4 hour duty cycle or in a control chamber. After
of OSNs expressing particular ORs using gene-targeted exposure, the exposed ester and the control ester both
mice. MOR23-GFP mice were exposed daily to Lyral and evoked less OSN synaptic output into fewer glomeruli
anatomical, molecular and physiological properties of these than prior to exposure, while aldehyde- and ketone-evoked
neurons were analyzed. The density of MOR23 neurons responses were unchanged. In a separate experiment, mice
decreased after odorant exposure while the level of mRNA underwent a similar baseline imaging session but were
for the receptor remained stable at the entire mucosa level. then differentially fear conditioned to associate 1 ester
To investigate molecular changes within individual OSNs, (the CS+) with shock while the other ester (the CS-) was
mRNA levels for olfactory signaling pathway compo- presented without shock. In a post-conditioning imaging
nents were quantitatively analyzed using qPCR on GFP- session the glomeruli that received OSN input driven by
containing neurons (7 per mouse). The levels of mRNAs the CS+ received larger synaptic inputs from OSNs, while
for CNGA2, PDE1C and MOR23 olfactory receptor were responses to the CS- and non-presented control odorants
higher in exposed OSNs compared to control. Using patch- were unchanged. Control mice that experienced only
clamp recordings on the dendritic knobs of MOR23 neu- odors or only shocks between imaging sessions exhibited
rons in an intact preparation we observed that exposed no changes in the response to any odorant. These data
OSNs displayed a lower detection threshold compared to provide surprising evidence that environmental changes
control OSNs while the dynamic range of the dose-response and emotional learning can change how OSNs respond
was broader. Responses of exposed neurons were also faster to olfactory stimuli. Acknowledgements: This work was
and shorter than the responses of control neurons. Postnatal supported by the National Institute on Deafness and Other
odorant exposure induces molecular and physiological Communication Disorders (R00 DC009442 to JPM).
#51 SYMPOSIUM: EXPERIENCE DRIVEN #53 SYMPOSIUM: EXPERIENCE DRIVEN

PLASTICITY OF THE OLFACTORY SYSTEM PLASTICITY OF THE OLFACTORY SYSTEM
Understanding Plasticity in the Olfactory Intrabulbar Map Olfactory Experience Shapes Insect Olfactory Centres
Leonardo Belluscio Jean-Marc Devaud
National Institutes of Health / NINDS Bethesda, MD, USA Research Center on Animal Cognition, Université Paul
Sabatier Toulouse, France
In the mammalian olfactory system sensory neurons pro-
ject their axons to the surface in the olfactory bulb generat- Insects provide excellent models to study how neural
ing a pair of glomerular maps that reflect odorant receptor networks dedicated to olfactory processing are formed during
identity. These maps are further connected through a set development, and how they work in the adult. However, their
of reciprocal intrabulbar projections that are mediated by organisation is not fixed once development is achieved. On
tufted cells that specifically link iso-functional odor columns the contrary, as in vertebrates, the connectivity and functional
to produce a second order map called the intrabulbar map. organisation of insect olfactory systems are not fixed: they
We have shown that intrabulbar projections are established exhibit clear plastic properties, as shown in various species
postnatally and undergo continuously refinement through over the recent years. In our work, we have been focusing on
an activity dependent process that has no critical period. the plastic changes affecting the anatomy of the olfactory
Here we present that both loss of olfactory sensory input centres as a consequence of olfactory experience, be it the
and broad odorant stimulation are capable of disrupting the mere exposure to environmental odorants or associative
intrabulbar map specificity, while re-introduction of normal learning and memory. In particular, we have been looking
activity restores the map to proper order. We also reveal that for structural rearrangements in two main olfactory centres
the regenerating interneurons are central to intrabulbar cir- known for their role in olfactory learning and memory in the
cuit plasticity and that proper connectivity depends specifi- insect brain: the antennal lobes and the mushroom bodies.
cally upon new neurons from the rostral migratory stream. The modular organization of these two neuropils allows
Together these data illustrate that olfactory bulb plasticity quantifying the changes affecting their structure in the brains
is a balance between activity, regeneration and remodeling. of animals submitted to different treatments. By doing so,
Acknowledgements: National Institute of Neurological and by focusing mostly on the honeybee (Apis mellifera) as a
Disorders and Stroke, Intramural Research Program. model species, we have been able to show that the formation
of long-term memories of previous olfactory experience is
associated with structural modifications in insect olfactory
networks. Interestingly, such modifications vary with the
#52 SYMPOSIUM: EXPERIENCE DRIVEN
nature of the experience undergone by the animal, and may
be considered as supports of olfactory memories. Thus, they
Long-term imaging of odor representations in awake mice are likely to contribute to the acquisition and retention of
Takaki Komiyama behavioural responses adapted to changing environments.
University of California, San Diego, CNCB, La Jolla CA

92093 USA #53.5 CLINICAL LUNCHEON: TASTE RECEPTORS
How are sensory representations in the brain influenced by IN GUT AND PANCREAS REGULATE ENDOCRINE
the state of an animal? Here we use chronic two-photon FUNCTION
calcium imaging to explore how wakefulness and experi- Robert F. Margolskee
ence shape odor representations in the mouse olfactory
Monell Chemical Senses Center, Philadelphia, PA, USA
bulb. Comparing the awake and anesthetized state, we show
that wakefulness greatly enhances the activity of inhibitory Many of the receptors and downstream signalling proteins
granule cells and makes principal mitral cell odor responses involved in taste detection and transduction are expressed
more sparse and temporally dynamic. In awake mice, brief also in intestinal and pancreatic endocrine cells where they
repeated odor experience leads to a gradual and long-last- underlie certain chemosensory responses. Intestinal endo-
ing (months) weakening of mitral cell odor representations. crine cells express T1r taste receptors, the taste G-protein
This mitral cell plasticity is odor specific, recovers gradu- gustducin, and several other taste transduction elements.
ally over months, and can be repeated with different odors. So too do pancreatic islet endocrine cells. Knockout mice
Furthermore, the expression of this experience- dependent lacking alpha-gustducin or the sweet taste receptor subunit
plasticity is prevented by anesthesia. Together, our results T1r3 have deficiencies in intestinal secretion of glucagon-like
demonstrate the dynamic nature of mitral cell odor repre- peptide-1 (GLP-1) and in the regulation of plasma levels of
sentations in awake animals, which is constantly shaped by insulin and glucose. Glucose-dependent insulin release from
recent odor experience. mouse pancreatic islets ex vivo is stimulated by sucralose
and other sweeteners. Islets from T1r3 knockout mice release non-volatile stimuli. Whether human airways can
insulin normally in response to glucose, but show no also detect volatiles is not known. However, human
enhanced release of insulin in response to non- caloric sweet- pulmonary diseases such as asthma have been linked to
eners. Thus, there appear to be two mechanisms for regulat- increased sensitivity of the airways to various volatile
ing insulin release from pancreas, one dependent on glucose, insults. We have identified several expressed canonical
glucose transporters and glucokinase, and another for sweet- olfactory receptors in human primary airway epithelia.
eners that involves T1r3 and other taste proteins. Only lim- Immunohistochemistry on lung sections and primary
ited studies with humans have been done in this area, but airway cultures to identified candidate olfactory sensory
it seems likely that “taste” signalling proteins in human gut cells in human airways, which can respond to volatiles
and pancreas also contribute to chemosensory responses in in culture. Using several well-established markers for
gut and pancreas to regulate glucose homeostasis. various pulmonary epithelial cell types we identified
the olfactory cells as pulmonary neuroendocrine cells
(PNEC). In humans, PNECs are morphologically distinct
#54 SYMPOSIUM: THE NEW ‘FACES’
cells of unknown function. We found that human PNECs
OF CHEMOSENSATION — UTILIZING express members of the olfactory receptor family and are
CHEMOSENSORY SIGNALING anatomically positioned to respond to inhaled volatile
PATHWAYS OUTSIDE THE CANONICAL chemicals. Further, human olfactory PNECs showed
CHEMOSENSORY ORGANS high levels of vesicular 5-HT and the peptide hormone
Symposium: The New ‘Faces’ of Chemosensation – Utilizing CGRP, further establishing the pulmonary neuroendocrine
Chemosensory Signaling Pathways Outside the Canonical system as olfactory sensory sentinels. Apical exposure of
Chemosensory Organs primary human airway cultures to volatile chemicals led
to the release of the neuroendocrine content of PNECs,
Yehuda Ben-Shahar1,2 indicating that adult human PNECs could act as olfactory
Washington University St. Louis, MO, USA, 2Washington
1 sensory cells. Since pulmonary tissues express diverse
University School of Medicine/Pulmonary & Critical Care serotonin and peptide receptors, these data indicate that
Medicine St. Louis, MO, USA human airway epithelia evolved a specialized group of cells
that can act autonomously in response to volatile chemical
In recent years, several exciting studies indicated that canon-
insults. These cells may represent the missing cellular and
ical mammalian chemosensory signaling pathways are also
physiological links between the exposure to environmental
likely to function outside the canonical chemosensory organs
volatiles and airway hypersensitivity observed in some
(taste and olfaction). These findings significantly expand
pulmonary diseases. Acknowledgements: NIH NIDCD
our field of view in terms of what constitutes a chemosen-
R03DC010244 and the Children’s Discovery Institute (St.
sory organ, and suggest that cell-autonomous chemorecep-
Louis)
tion, independent of the nervous system, is likely playing an
important role in health and disease, which includes diges-
tive, respiratory, and reproductive functions. This sympo- #56 SYMPOSIUM: THE NEW ‘FACES’
sium will present the current state of knowledge about the
OF CHEMOSENSATION — UTILIZING
possible extra-sensory functions of ‘taste’ and ‘olfaction’
signaling molecules in non-sensory tissues and cells. CHEMOSENSORY SIGNALING
PATHWAYS OUTSIDE THE CANONICAL
CHEMOSENSORY ORGANS
#55 SYMPOSIUM: THE NEW ‘FACES’
OF CHEMOSENSATION — UTILIZING Bitter Taste Receptors on Airway Smooth Muscle: a target
for novel bronchodilators
CHEMOSENSORY SIGNALING
PATHWAYS OUTSIDE THE CANONICAL Stephen Liggett
CHEMOSENSORY ORGANS University of South Florida Morsani College of Medicine.
Human Pulmonary Neuroendocrine Cells are Olfactory Tampa, FL
Sentinels There is an unmet need for treatments of the airway con-
Yehuda Ben-Shahar1,2 striction that occurs in asthma. Currently, beta-agonists
are the only class of direct bronchodilators that are in use.
Washington University St. Louis, MO, USA, 2Washington
1
We unexpectedly found T2Rs (particularly subtypes 10, 14,
University School of Medicine/Pulmonary & Critical Care
31) expressed on human airway smooth muscle (ASM).
Medicine St. Louis, MO, USA
Activation by T2R agonists causes significant and revers-
Previous work indicated that mammalian pulmonary ible ASM relaxation in human and mouse airways, isolated
ciliated epithelial cells act as chemosensory cells to human ASM cells studied by magnetic twisting cytometry
and Fourier transform traction maps, and in mice in vivo. #58 SYMPOSIUM: THE NEW ‘FACES’
Signaling was sensitive to inhibitors of betagamma, PLC, OF CHEMOSENSATION — UTILIZING
and the IP3 receptor, and was associated with membrane CHEMOSENSORY SIGNALING
hyperpolarization and an increase in SR-released intracel- PATHWAYS OUTSIDE THE CANONICAL
lular [Ca2+] within a sequestered pool restricted to the cell
CHEMOSENSORY ORGANS
surface. One channel that appears to be involved is BKca,
but there may be others. In the ovalbumin-sensitized mouse Detection Of Irritants And Bacterial Metabolites Via The
model of asthma, the inhaled T2R agonist quinine was much Taste Transduction Cascade In Solitary Chemosensory Cells
more efficacious than the inhaled beta agonist albuterol in Of The Nasal Cavity
reversing airflow obstruction. Given the large number of
Thomas Finger1,3, Sue Kinnamon2,3, Vijay Ramakrishnan2,
known bitter tastants, there is an opportunity to discover
Marco Tizzano1,3
non-toxic T2R agonists for inhalation for the treatment of
obstructive lung disease. 1
Dept. Cell & Devel. Biol., Univ Colo. Med. Sch. Aurora, CO,
USA, 2Dept. Otolaryngology, Univ Colo. Med. Sch. Aurora,
CO, USA, 3Rocky Mountain Taste & Smell Center Aurora, CO,
#57 SYMPOSIUM: THE NEW ‘FACES’ USA
OF CHEMOSENSATION — UTILIZING Airways are continually assaulted by harmful compounds
CHEMOSENSORY SIGNALING carried on the incoming airstream and by the potentially
PATHWAYS OUTSIDE THE CANONICAL pathogenic bacterial populations. We have shown that the
CHEMOSENSORY ORGANS nasal epithelium of rodents houses a population of trigem-
inally-innervated solitary chemosensory cells (SCCs) that
Genetics of the bitter taste receptor T2R38 underlie
express T2R taste receptors along with their downstream
susceptibility to upper respiratory infection
signaling components crucial for detection and response
Noam A. Cohen1,2, Robert J. Lee1, Danielle R. Reed3, Peihua to these deleterious agents. SCCs are distributed across
Jiang3, Gary K. Beauchamp3 much of the nasal respiratory epithelium in rodents, but
are especially concentrated along curved surfaces facing
1
University of Pennsylvania/Otorhinolaryngology - Head major airway conduits. In the upper respiratory passage-
and Neck Surgery Philadelphia, PA, USA, 2Philadelphia VA ways, the SCCs express nearly all T2Rs tested as well as
Medical Center/ Surgery Philadelphia, PA, USA, 3Monell some T1R family members. Downstream signaling com-
Chemical Senses Center Philadelphia, PA, USA ponents also are expressed by SCCs including: gustducin,
PLCß2 and TrpM5. Functionally, SCCs respond rapidly
Innate and adaptive defense mechanisms protect the to bitter ligands (e.g. denatonium 10mM) as well as bac-
respiratory system from attack by microbes. Here, terial signaling molecules including the acylhomoser-
I describe our recent studies that demonstrate that the ine lactones produced as quorum sensing molecules by
bitter taste receptor T2R38 is expressed in the human Pseudomonas species. Activation of the SCCs by these
upper respiratory epithelium and is activated by acyl- compounds evokes neurally-mediated protective reflexes,
homoserine lactone quorum sensing molecules secreted by involving both apnea and local neurogenic inflamma-
Pseudomonas aeruginosa and other gram-negative bacteria. tion. These responses are absent in both gustducin and
T2R38 regulates human upper airway innate defenses TrpM5 knockout mice implicating the taste transduction
through nitric oxide production, resulting in stimulation cascade as a crucial component of responses to these
of mucociliary clearance and direct antibacterial effects. substances. Moreover, the local inflammatory responses
Moreover, common polymorphisms of the TAS2R38 were absent after chemical ablation of the nociceptive
gene are linked to significant differences in the ability of fibers of the trigeminal nerve showing the necessity for
upper respiratory cells to clear and kill bacteria. Lastly, innervation. More recently we have tested the possibil-
TAS2R38 genotype correlates with human sinonasal ity that similar SCCs are present in the human nose and
gram-negative bacterial infection. These data suggest now report (S. Cooper AChemS 2013) the presence of
that T2R38 is an upper airway sentinel in innate defense, elongate TrpM5+ microvillous cells within the sinonasal
and that genetic variation that contributes to human epithelium in humans. Whether similar SCCs underlie
individual differences in ability to taste phenylcarbamide epithelial defense systems in humans remains to be deter-
(PTC) and related molecules also contributes to individual mined. Acknowledgements: Supported by NIDCD grants
differences in susceptibility to respiratory infection. to T. Finger, SC Kinnamon (RO1 DC009820), M Tizzano
Acknowledgements: P30DC011735 R01DC004698 (R03 DC012413) and the Rocky Mountain Taste & Smell
P50DC000214 R01DC010842 Center (P30 DC04657)
#59 SYMPOSIUM: THE NEW ‘FACES’ human Tas2r bitter taste receptors even though the number
OF CHEMOSENSATION — UTILIZING of cognate compounds discovered for the human TAS2Rs
CHEMOSENSORY SIGNALING was larger. This is surprising since the mouse genome
PATHWAYS OUTSIDE THE CANONICAL encodes 30% more bitter receptors than the human genome
does. Notably, the two species usually detect the same com-
CHEMOSENSORY ORGANS
pounds with non- orthologous Tas2rs. Both species possess
Block of taste genes leads to male sterility generalists, moderately tuned Tas2rs and specialists accord-
Bedrich Mosinger, Kevin M. Redding, Rockwell M. Parker, ing to their number of cognate bitter compounds. However,
Robert F. Margolskee a larger fraction of specialists occurs in mice, proposing that
the luxury of having specific receptors for solitary bitter
Monell Chemical Senses Center Philadelphia, PA, USA compounds in a species is supported by a greater number of
The male infertility rate in the developing world is increasing. TAS2R genes.
About 7% of men are infertile for unknown reasons, while hav- Genetic labeling and in situ hybridization experiments
ing normal hormone levels. We found that two gene products visualized the populations of bitter-sensing cells in mice and
(T1R3 and gustducin) originally found in taste chemosensa- man. They express the complete repertoires of Tas2r genes
tion are expressed in haploid spermatids and their absence or but individual cells express only limited subsets. This is sup-
block leads to male sterility. The pathology shows an arrest in ported by genetic ablation in mice of the cells for Tas2r131
spermatid development with numerous giant and exfoliated which extinguished only ~50% of the bitter cell population.
cells in testicular tubules, oligospermia and immotile sperm. Oral administration of bitter compounds to mice excites spe-
To study this phenomenon we produced a mouse model cifically 200–400 neurons in the gustatory part of the nucleus
expressing a humanized form of T1R3 that can be inhibited of the solitary tract as indicated by induction of immediate
by human-specific inhibitors. Some of these inhibitors are early gene. These gustatory neurons respond differently to
phenoxy compounds developed into common medications different oral bitter stimuli. Finally, we found that mice with-
and agricultural chemicals. A very effective, yet reversible, ste- out Tas2r131 cells avoid several bitter compounds less than
rility is achieved by treatment with the phenoxy compound controls, whereas they are indistinguishable from controls in
clofibrate in this model with humanized T1R3 receptor in their avoidance of denatonium benzoate.
the background of gustducin null allele. Because both T1R3 Our data demonstrate that bitter sensing cells and their
and gustducin affect cAMP levels, we hypothesize that their gustatory target neurons are functionally distinct forming
absence impairs function of CREM (cAMP response modu- the basis for variable behavioral responses to different bitter
lator protein) a transcriptional master gene indispensable for chemicals which could be of relevance for ingestive behavior.
spermatid development and EPAC (exchange protein activated
by cAMP) a Rap 1 activator required for cell adhesion. We fur-
ther hypothesize that even low levels of chemicals and/or medi- #P1 POSTER SESSION I: MULTIMODAL
cations acting on T1R3 and gustducin, possibly in combination RECEPTION; CHEMOSENSATION AND DISEASE;
with other drugs, can negatively affect human male fertility. OLFACTION PERIPHERY
Acknowledgements: R21 DC007399 - NIH/NIDCD, R01
Meteorological Vaticination through Phantosmia:
DC003155 - NIH/NIDCD, P30DC011735 - NIH/NIDCD.
A Case Study
Gurprit Bains, Salvatore Aiello, Alan Hirsch
#60 IFF LECTURE: BITTER TASTE IN MICE Smell & Taste Treatment and Research Foundation Chicago,
AND MAN IL, USA
Bitter Taste in Mice and Man Introduction: Linkage of weather to chemosensory hallucina-
Wolfgang Meyerhof tions has heretofore not been reported. Methods: Case Study:
A 64 yo M presents with five years of intermittent noxious
Department of Molecular Genetics, German Institute of skunk-onion excrement phantosmia, lasting for hours. If
Human Nutrition, Nuthetal, Germany severe, he tastes the same odor. It is exacerbated by coughing
Bitterness is elicited by numerous structurally diverse and nasal congestion, and alleviated with sleep, nasal irriga-
molecules which usually act as repellents allowing us to avoid tion, alprazolam, occluding nostrils, assuming Moffitt’s posi-
intake of harmful food. However, we adapt to and even prefer tion, snorting salt water, blowing nose, and holding breath.
the bitterness of some foods and beverages. To elucidate these When eating or sniffing, the actual flavors replace the phan-
phenomena my laboratory investigates the receptors and cells tosmia. Since onset, he noted the intensity and frequency of
for bitter tasting compounds in man and mice. the phantosmia forecasted the weather. Two hours before a
Functional assays with >100 chemicals demonstrate similar storm, the phantosmia intensifies from a level 0 to a 7–10,
molecular receptive ranges for the repertoires of murine and which persists through the entire thunderstorm. Twenty years
prior, he noted the ability to forecast the weather, based on treatment for those who suffer from dizziness or in stability.
pain in a torn meniscus, which vanished after surgical repair. Further testing on those suffering from dizziness is warranted
Results: Olfactory testing revealed hyposmia: QSIT 1; ETOH
Sniff Test 3cm; BSIT 8; PST 2; Odor Memory Test: 2–10 sec., #P3 POSTER SESSION I: MULTIMODAL
2–30 sec., 0–60 sec.; UPSIT – 27 R, 8 L, Sniffin’ Sticks – RECEPTION; CHEMOSENSATION AND DISEASE;
Threshold: L <1, R <1, dirhinous <1, Discrimination: L 9, OLFACTION PERIPHERY
R 8, dirhinous 10; Identification: L 8, R 6, dirhinous 6; Sniff
Magnitude Test: 1.02 – 1.12 (Anosmia); PEA Testing: L= - Superficial Siderosis-Induced Anosmia
5.0 R > -2.0. Conclusion: Decrease in barometric pressure Alan R Hirsch, Sanford Sherman, Gul Hwang
may precipitate phantosmic synestesthia or reduces already Smell & Taste Treatment and Research Foundation Chicago,
hyposmic olfactory ability, serving to disinhibit phantosmia. IL, USA
Atmospheric changes may induce contractions on a scarred OBJECTIVE: Superficial siderosis, a rare condition that
olfactory nerve, heightening ectopic discharge. Menacing results from the deposition of hemosiderin in the central
weather may impair mood, enhancing awareness of intero- nervous system, has been reported to cause anosmia in 17%.
ceptive bodily symptoms and somatic sensory amplifica- METHODS: A 49-year old female patient with superficial
tion. Misattribution error may have occurred by expectation siderosis and anosmia is presented. RESULTS: Chemosensory
effect or selective attention. This is the first reported case of function was evaluated using ETOH Sniff Test; Quick Smell
weather-induced exacerbation of phantosmia. Identification Test; Brief Smell Identification Test; Pocket
Smell Test; University of Pennsylvania Smell Identification
Test, Extended Sniffin’ Stick Tests of threshold, discrimina-
#P2 POSTER SESSION I: MULTIMODAL tion and identification; Suprathreshold Amyl Acetate Odor
RECEPTION; CHEMOSENSATION AND DISEASE; Hedonic and Intensity Tests, Monorhinous Phenylethyl
OLFACTION PERIPHERY Alcohol Smell Threshold Test; Retronasal Smell Testing,
The Effects of Aroma of Baked Cinnamon Bun on Stability Accusens Taste Tests, Taste Quadrant Testing, and PROP
paper. Abnormalities were noted in mono and dirhinous
Alan Hirsch, Salvatore Aiello detection, discrimination, identification, hedonics, intensity
Smell & Taste Treatment and Research Foundation Chicago, perception, retronasal smell, taste threshold to HCl, and taste
IL, USA quadrant to sucrose and quinine. Besides anosmia and hypo-
geusia, she had mild sensorineural hearing loss and chronic
Objectives: Dizziness or instability affects almost 70 million
bifrontal throbbing headache, which was diagnosed as pseu-
Americans, and is of multimodal origin. Odors can influence
dotumor cerebri. Brain MRI revealed hemosiderin deposition
many physiological systems including, the automatic nervous
in the bilateral cerebellar hemispheres, left Sylvian fissure,
system, nociceptive, visual, and somesthetic; all of which play
and the anterior poles of bilateral temporal lobes. Lumbar
a role in dizziness. No studies have demonstrated the effects
puncture revealed increased pressure and blood in cerebro-
of odors on dizziness. This study addresses if a hedonically
spinal fluid. CONCLUSIONS: The chemosensory loss of the
pleasant odor, baked cinnamon bun (BCB), can impact
patient is postulated to be due to deposition of hemosiderin
stability. Methods: 11 normosmic (3/3 Q-SIT) subjects, aged
on the olfactory nerve. In superficial siderosis, anosmia or
20–58, 3 males, 8 females, underwent a test of stability while
hyposmia is common, but olfactory function testing is rarely
wearing blank surgical masks or surgical masks impregnated
undertaken. Earlier case reports of superficial siderosis have
with BCB (IFF), in a counter balanced order. Subjects
not described detailed chemosensory tests. This is the first
completed the Single Leg Balance Test using the Nintendo
report of superficial siderosis-induced chemosensory deficit
Wii FitTM software in combination with the Nintendo Balance
with presentation of a set of objective chemosensory testings.
BoardTM as per published protocol. Subjects were tested on the
non-dominant leg (the opposite of the leg used to kick a ball).
Results: 10 subjects reported positive hedonics to the odor #P4 POSTER SESSION I: MULTIMODAL
of BCB. Order of presentation had no effect on the outcome RECEPTION; CHEMOSENSATION AND DISEASE;
(p=.96). Gender does have an effect on the outcome, with OLFACTION PERIPHERY
higher stability percentage in females (66.7 vs. 65.6, p=.0009). Postviral Hyposmia with Transient Improvement by
However, the BCB-versus- blank effect is not different between Spontaneous Yawning
men and women. Stability percentage is significantly higher
Alan R Hirsch, Jason Gruss, Gul Hwang
using BCB compared to blank; Stability percentage 67%
using BCB, 60% with un-odorized mask, with each person Smell & Taste Treatment and Research Foundation Chicago,
serving as their own control (p=.02). Conclusions: BCB odor IL, USA
improved balance significantly in our healthy test subjects. The Objective: To describe a patient with hyposmia whose
use of hedonically favorable BCB aroma may have utility in olfactory ability returns upon yawning. Method: 61yo male,
three years ago noted nasal congestion, followed by loss subjects verified they had knowledge of what these flavors
of smell and taste. Sinus CT showed opacification of the should taste like. Results: Significant differences(p<0.05)
ethmoidal and an air- fluid level in the right maxillary sinus. were found between the means of: cappuccino 5.3, popcorn
After treatment, sinus CT, MRI, and fiberoptic endoscopy 5.2, mint chocolate chip ice cream 5.1, cinnamon 4.4, grape
were normal, but smell ability remained at 5%. He is able 4.2, mango chili 4.1, bubblegum 4.1, Tabasco 4.0, 7-UP 3.6,
to smell flowers and gasoline if held close. Immediately and cotton candy 2.9. Conclusions: Results confirmed in
after yawning he was able to smell for a second. With small normosmic children Mozell’s finding that coffee had the
yawns 20% returns; 100% with large yawns. Two years greatest retronasal component and Bartoshuk’s observa-
ago transient phantosmias began of rubber or chemicals. tion that cappuccino jelly beans are the best testing agent
His taste is 1–2%. He can taste Chinese mustard and red to assess retronasal smell
pepper. Results: Chemosensory testing indicated hyposmia
and hypogeusia. Q-SIT 2; Sniff Magnitude with Sniff
Magnitude Ratio of .87; Alcohol Sniff Test 11 cm; Sniffin’ #P6 POSTER SESSION I: MULTIMODAL
Stick Threshold L <1, R <1, bilateral <1; Discrimination RECEPTION; CHEMOSENSATION AND DISEASE;
L 4, R 4, bilateral 6; Identification L 4, R 5, bilateral 7; OLFACTION PERIPHERY
UPSIT R 19, L 18; Odor Memory Test 10 sec 3, 30 sec 4,
60 sec 2, total 9/12; Smell Threshold with PEA L > - 2.0, Examining the Relationship Between Subjective Smell Loss
R > - 2.0; Suprathreshold Amyl Acetate Odor Intensity - Ratings and Olfactory Test Performance in Traumatic Brain
normal at high intensity; Suprathreshold Amyl Acetate Injury (TBI) Patients Pursuing Injury Litigation
Odor Hedonics - normal. Spatial Taste Test - weakness on Elissa C. McIntosh1, Claire Murphy1,2
the whole tongue and palate to NaCL and Sucrose, right 1
San Diego State University San Diego, CA, USA, 2SDSU/UCSD
anterior and posterior tongue to citric acid and quinine
Joint Doctoral Program in Clinical Psychology San Diego, CA,
hydrochloride. PROP disc - normal. Fungiform papillae
USA
L 22, R 18. Retronasal smell: with nose clip 1/10, without
nose clip 4/10. Dirhinous PEA Threshold Testing -2.5; After In this study, we investigated the relationship between smell
Polite Yawning Technique > -2.0; After Induced natural loss ratings and olfactory test performance in 45 traumatic
yawn > -2.0. Conclusion: Yawning may improve olfaction brain injury (TBI) patients. The objectives of the study were
by enhancing nasal airflow. The polite yawning technique (1) to determine if the relationship between ratings and per-
should be considered to be part of the evaluation in those formance differed among olfactory tests, and (2) to deter-
who complain of chemosensory dysfunction. mine if the relationship between ratings and performance
was significantly altered in TBI patients involved in litiga-
tion. Each patient completed Olfactory Threshold and Odor
#P5 POSTER SESSION I: MULTIMODAL Identification (ID) testing. Further, each patient rated their
RECEPTION; CHEMOSENSATION AND DISEASE; smell loss on a scale from 0 (no disability) to 100 (maximum
disability). When examining all TBI patients, regardless of
OLFACTION PERIPHERY
litigation status, regression analyses in each test showed a
Tests of Retronasal Smell in Children: Which Flavored Jelly significant relationship between ratings and performance.
Bean Works Best? As ratings increased (more disability), performance on both
Noah Hirsch1, Richard Bone2, Alan R Hirsch1 tests decreased. However, when accounting for litigation, we
found differences between groups. For patients not involved
1
Smell & Taste Treatment and Research Foundation Chicago, in litigation, we found significant relationships between odor
IL, USA, 2Christ Advocate Medical Center Oak Lawn, IL, USA ID and ratings (p<.001), and between threshold and ratings
Objective: In 13 adults, Mozell et al (1969) found flavor (p=.01). Conversely, for patients involved in litigation, we
dependent effects of different foods of retronasal smell on did not find significant relationships between odor ID and
identification, but did not address intensity. Engen (1982) ratings (p=.072), or between threshold and ratings (p=.224).
posited that the olfactory role in identification differs from From these results, we can conclude that there is a relationship
intensity assessments. We looked to determine, in chil- between ratings and test performance when litigation status
dren, the retronasal component of intensity of flavored is not considered. However, this relationship is weaker (not
jelly beans. Methods: Forty two, 12 or 13 year olds, were significant) for TBI patients involved in litigation and these
screened with the Brief Smell Identification test. Twenty patients are not as accurate in rating their olfactory deficits.
five (17 girls, 8 boys) scored 3/3 and underwent an assess- Though further study is warranted, this finding might point
ment of intensity, on a 10 point visual analog scale, with to a problem with malingering in this specific patient popula-
and without nose clips, of 10 different Jelly Belly jelly tion, and suggests a need for more comprehensive olfactory
beans. The mean differences were determined. Each jelly testing when assessing TBI patients. Acknowledgements:
bean flavor was provided to subjects prior to testing and Supported by NIH grant R01AG04085 to CM.
#P7 POSTER SESSION I: MULTIMODAL pH, adrenal level, and cortisol level, as well as general
RECEPTION; CHEMOSENSATION AND DISEASE; health indices, of 107 participants who were either breast-
OLFACTION PERIPHERY fed or not breast-fed as an infant. Data were subjected to
independent t-tests, with group (breast-fed vs. non-breast-
Prenatal Alcohol Exposure Impairs Olfactory Function fed) serving as the independent measure and physiologi-
in Humans cal measurements (salivary pH, adrenal level, and cortisol
Claire Murphy1,2, Emily S. Bower2, Jacquelyn Szajer1, Sarah level) and general health indices serving as dependent
N. Mattson1,2, Edward P. Riley1,2 measures. A significant pH difference was found between
San Diego State University San Diego, CA, USA, 2SDSU/
1 groups, with breast-fed individuals having more acidic
UCSD Joint Doctoral Program in Clinical Psychology San saliva, t(106)= −2.24, p=.03. Differences between groups
Diego, CA, USA were also found for measures of health-consciousness
[t(64)= −1.78, p=.08], health-status [t(64)=2.01, p=.05],
Children with histories of prenatal alcohol exposure often
healthiness-prevention [t(64)=-1.65, p=.10], and health-
suffer devastating consequences. Fetal alcohol syndrome
depression [t(64)= −1.79, p=.08]. Individuals that were
(FAS) is diagnosed by key facial features, growth deficits and
breast-fed showed higher health consciousness, higher
CNS anomalies. Children with histories of prenatal alcohol
health status, higher health prevention, and lower health
exposure (EA) who do not exhibit all of the features of FAS
depression than non-breast-fed individuals. Implications
also often have behavioral and cognitive impairments, though
of these results may influence mothers to consider their
characterizing the EA phenotype remains challenging.
child’s future general health in adulthood when deciding
Many of the regions impacted by EA (orbitofrontal cortex,
to breast-feed or use formula milk.
medial temporal lobe, limbic areas) are critically involved in
processing olfactory information, thus olfactory measures
may contribute to better identification of EA. Although
there are no publications reporting results of testing human #P9 POSTER SESSION I: MULTIMODAL
olfactory performance in FAS or EA, studies of FAS in RECEPTION; CHEMOSENSATION AND DISEASE;
animal models suggest decreased olfactory bulb volume OLFACTION PERIPHERY
and the potential for impaired olfactory function. Here we Alliaceous Migraines
compared children with EA to typically developing controls
(NC) (N = 11/group). It was hypothesized that children in the Alexander Roussos, Alan Hirsch
EA group would perform more poorly than the NC group on Smell & Taste Treatment and Research Foundation Chicago,
the San Diego Odor Identification Test. The results showed IL, USA
that children with EA were significantly impaired in odor
Objective: To report a migraineur with osmophobia and
identification (M = 5.48, SE = .45) compared to typically
trigger to garlic and onion Methods: 32-year-old woman; five
developing age-matched controls (M = 7.16, SE = .45),
years ago felt nasal pruritis upon eating red onion dip. Shortly
F(1,19) = 6.83, p <.05, partial δ2 = 0.26. This is the first
thereafter, mere aroma of raw onions caused the sensation of
report of impaired odor identification in humans with EA.
her throat closing and panic attack. Over time, she developed
The results support compromised olfactory performance
headaches upon exposure to onions and garlic; preceded by
in EA, and suggest that further research is warranted to
aura of fortification spectra and visual entopia. Followed by
identify the mechanisms underlying these deficits, the
a bipareital, crushing level 10/10 headache, burning eyes and
integrity of brain areas that are involved, and to determine
nose, lacrimation, perioral paresthesias, generalized pruritis,
whether olfactory performance can contribute to better
nausea, fatigue, sore throat, dysarthria, confusion, dyspnea,
identification of children at risk for behavioral and cognitive
palpitations, presyncopal sensations, hand spasms, tongue
deficits. Acknowledgements: Supported by NIH grants
soreness, neck pain, phonophobia, and photopobia. These
R01AG004085-25 to CM and R01AA010417-14 to EPR.
would persist for one hour after leaving the aroma. She is
unresponsive to medication and wears a surgical mask when
#P8 POSTER SESSION I: MULTIMODAL out. She also has chemosensory complaints: dysosmias every
RECEPTION; CHEMOSENSATION AND DISEASE; few months; phantosmias of food or cleaning products every
OLFACTION PERIPHERY month for a minute of level 5/10 intensity; palinosmia of
Differences in Salivary pH and General Health Status onion or garlic odor for 30 minutes after exposure; metallic
Among Individuals Who Were and Were Not Breast-fed palinageusia after eating with metal utensils for a few
minutes. Results: Neuro exam normal except for bilateral
Bryan Raudenbush, August Capiola, Amanda Schultz
positive Hoffman reflexes. Chemosensory testing: QSIT 3/3,
Wheeling Jesuit University Wheeling, WV, USA BSIT 12/12. MRI and CT with and without contrast normal.
Past research shows positive effects of breast-feeding dur- Allergy skin test was positive for garlic and onion. Nose plug
ing infancy. The present study investigated adult salivary and counter stimulation with peppermint prevented the onset
of migraine. Conclusion: This is the first report of migraines #P11 POSTER SESSION I: MULTIMODAL
triggered by more than one alliaceous compound in the RECEPTION; CHEMOSENSATION AND DISEASE;
same individual. Possible mechanisms include odor induced: OLFACTION PERIPHERY
emotional change; vasomotor instability; trigeminal induced
neurogenic inflammation; and allergic response. In alliaceous The WUTC Odor Threshold Test: Evaluating Olfactory
and odor-induced migraines, a trial of counter stimulation Ability Using Signal Detection Theory
and nose plugs is warranted. William Tewalt, Irene N Ozbek, McKinney Jessica, Santiago
Manuel, Biderman Michael
University of Tennessee at Chattanooga Chattanooga,
#P10 POSTER SESSION I: MULTIMODAL TN, USA
RECEPTION; CHEMOSENSATION AND DISEASE; A new odor detection threshold test (WUTC) was devel-
OLFACTION PERIPHERY oped, using signal detection theory, to address the need for
Dysautonomia and Chemosensory Dysfunction establishing a measure of consistency of subjects’ responses,
and the need for introducing different odorants to address
Noorussabah Shaikh1, Alan R. Hirsch1,2,3,4
different clinical/ research questions. The method of admin-
1
Smell and Taste Treatment and Research Foundation istration used is an improvement over previous methods of
Chicago, IL, USA, 2Mercy Hospital and Medical Center/ testing because the likelihood of desensitization to a single
Department of Medicine Chicago, IL, USA, 3Rush University odor over time is minimized. Odorants were chosen on the
Medical Center/ Department of Neurology Chicago, IL, USA, basis of the possible link of the detection of specific odor-
4
Rush University Medical Center/ Department of Psychiatry ants to known disease state. For example, using random
Chicago, IL, USA presentation, 5 different odors were presented at 9 different
Introduction: Myriad disorders associated with dysau- levels of concentration twice to a subject with End Stage
tonomias also display chemosensory dysfunction. Given Renal Disease (ESRD). Blanks were presented 9 times also.
this overlap, assessments of autonomic dysfunction Total administration time was 38 minutes. The subject was
amongst patients of a specialized chemosensory clinic 100% consistent for isoamyl acetate, 89% percent consist-
were performed. Methods: 25 consecutive patients at a ent for P-cresol, and 44% consistent for vanillin and blanks,
chemosensory clinic were approached to participate in this i.e. the consistency was less than chance suggesting that the
IRB approved study. 22 consented. Average age 51 years subject was guessing. This result is consistent with the pre-
(range 22 – 69) 8 males and 14 females, with diagnosis of diction that P-cresol may block the detection of vanillin in
hyposmia/ anosmia (18), dysosmis (3), phantosmia (6), ESRD patients. Acknowledgements: William H Wheeler
palinosmia (4), hypogeusia/ ageusia (15), dysgeusia (9), Foundation
phantogeusia (9), burning mouth syndrome (4). All under-
went the Quick Smell Identification Test (QSIT), and the #P12 POSTER SESSION I: MULTIMODAL
Survey of Autonomic Symptoms (SAS) for symptoms RECEPTION; CHEMOSENSATION AND DISEASE;
and Total Impact Score (TIS) for severity. Results: Our OLFACTION PERIPHERY
patients demonstrated fewer autonomic symptoms than
either those with autonomic disorders or even published Compensation Gone Awry: Conditions Inducing Regional
controls. There was a significant difference in SAS score in Oral Sensory Loss May Elevate Obesity Risk
the group as a whole (1.82) compared to published control Derek J Snyder1,2, Linda M Bartoshuk1,2
SAS of ≤3.0 (p= 0.003) and of the group as a whole (4.32) 1
Center for Smell and Taste, University of Florida Gainesville,
compared to TIS of controls ≤ 7.0 (p< 0.006). No signifi-
FL, USA, 2Community Dentistry, University of Florida
cance difference between QSIT score and either SAS or TIS
Gainesville, FL, USA
was observed (p>0.1). Discussion: This lack of autonomic
dysfunction in those chemosensory disorders may reflect Oral afferent nerves innervate portions of the mouth, convey
a sampling error. Those with autonomic dysfunction may unique arrays of information, and take different paths to the
be so burdened by their primary illness they did not seek brain. As such, certain health conditions predict specific
care for their chemosensory problems, if they even recog- patterns of regional oral sensory loss: Severe childhood ear
nized them at all. Alternatively, the SAS and TIS may not infections (otitis media, OM) damage the chorda tympani
be valid in this patient group, while physiologic autonomic (CT) and block anterior taste cues, while tonsillectomy
testing might demonstrate dysfunction. A larger sample damages the glossopharyngeal nerve (IX) and blocks
size may have revealed dysautonomia. Conclusion: Lack of posterior taste/tactile cues. These local effects disinhibit
autonomic symptoms were seen in those with chemosen- intact oral sensations; for example, CT block elevates
sory dysfunction. Further investigation of such a connec- IX, trigeminal, and whole-mouth intensity, particularly
tion is warranted. in supertasters of 6-n-propylthiouracil. Although this
compensatory mechanism often mitigates real- world (i.e., sex. Spatial taste testing (N=287) revealed significant loss
whole-mouth) perceptual deficits, our recent data implicate of taste at VII with intensifications in flavor and oral touch
it in a more subtle trend toward long-term obesity risk. In a (e.g., fat) in those subjects with intact IX. A third population
laboratory study (N = 301), individuals with medical histories is of special interest: athletes in contact sports like boxing or
indicating either CT damage (i.e., OM) or IX damage (i.e., football who tend to gain weight in retirement. We suggest
tonsillectomy) show elevated whole-mouth taste, oral burn that food preferences should be examined in head trauma to
and viscosity, and retronasal olfaction (RO). In survey data determine how much weight gain can be attributed to taste
from a larger sample (N = 6584), such individuals also show damage with resulting sensory and palatability alterations.
increased high-fat food avidity and body mass. Consistent Acknowledgements: DC283, DC8613 and DC8620
with reports linking RO to oral sensation, those with both OM
and tonsillectomy show reduced whole-mouth taste and RO,
revealing compensatory limits following extensive loss. While #P14 POSTER SESSION I: MULTIMODAL
this model remains exploratory – it requires verification in a RECEPTION; CHEMOSENSATION AND DISEASE;
single sample, and shifts in obesity risk with whole-mouth OLFACTION PERIPHERY
gain vs. loss remain to be seen – relative intensity changes
among flavor components appear sufficiently robust to Atherosclerosis and Decline in Odor Identification
influence high-fat intake. Overall, oral disinhibition sustains Carla R. Schubert1, Karen J. Cruickshanks1,2, Mary E. Fischer1,
whole-mouth taste and flavor perception by moderating Guan-Hua Huang3, Barbara E. Klein1, Ronald Klein1, James
the impact of limited spatial loss, but the strength of this S. Pankow4, Nathan Pankratz5, Alex Pinto1
effect may shape long-term food choice and dietary health. 1
University of Wisconsin/Department of Ophthalmology &
Acknowledgements: NIDCD (DC 00283)
Visual Sciences Madison, WI, USA, 2University of Wisconsin/
Department of Population Health Sciences Madison, WI,
USA, 3National Chiao Tung University/Institute of Statistics
#P13 POSTER SESSION I: MULTIMODAL
Hsinchu, Taiwan, 4University of Minnesota/Division of
RECEPTION; CHEMOSENSATION AND DISEASE; Epidemiology and Community Health Minneapolis, MN,
OLFACTION PERIPHERY USA, 5University of Minnesota/Laboratory Medicine and
Head Trauma, Taste Damage and Weight Gain Pathology Minneapolis, MN, USA
Linda M. Bartoshuk1, Susan Marino2, Derek J. Snyder1, Olfactory impairment is common in older adults although
Jennifer J. Stamps1 awareness of impairment is low, suggesting that olfactory
function may decline slowly over time. We evaluated factors
University of Florida Gainesville, FL, USA, 2University of
1
associated with a 5-year decline in odor identification in the
Minnesota Minneapolis, MN, USA
Beaver Dam Offspring Study, a longitudinal cohort study of
Severe brain injuries associated with weight gain have been adults aged 21–84 years at baseline (BOSS1; 2005–2008). The
reported in children (Jourdan et al. 2012; Norwood et al. 8-odorant San Diego Odor Identification Test (SDOIT) was
2010) and adults (Henson et al. 1993); one source of the administered at the baseline and 5-year follow-up (BOSS2; 2010–
weight gain is believed to be metabolic (e.g., hypothalamic 2013) examinations. Decline in odor identification was defined
dysfunction). Head trauma has long been known to affect as a decrease in SDOIT score ≥2 from BOSS1 to BOSS2; no
taste (Sumner 1967; Costanzo and Zasler 1991; Solomon change was defined as a difference between BOSS1 and BOSS2
et al. 1991). Our recent data suggest that taste damage from scores of ≤1. In preliminary analyses of the first 2195 participants
otitis media or tonsillectomy is associated with enhanced pal- with SDOIT data at BOSS1 and BOSS2, 3.1% had a decline
atability of energy dense foods and weight gain (see Snyder in SDOIT score. Those with a decline in odor identification
& Bartoshuk poster). We presented a model suggesting were more likely to be older (Odds Ratio (OR)=1.58, 95%
that damage to taste nerves VII or IX could intensify non- Confidence Interval(CI)=1.40, 1.79, per 5 years of age) than
taste oral sensations centrally possibly altering palatability those with no change in score. In age- and sex-adjusted models,
(Bartoshuk et al. 2012). Of special interest, the consequences baseline current smoking (OR=2.18, 95%CI=1.08, 4.37, vs
of damage to either VII or IX depended on the status of never), carotid artery intima media thickness(IMT)(OR=1.17,
the other nerve. Central intensifications only occurred when 95%CI=1.01, 1.36, per 0.1mm), number of carotid artery sites
the damage was restricted to one nerve. The present study (range 0–6) with plaque (OR=1.39, 95% CI=1.13, 1.70) and
extends these conclusions to taste damage resulting from report of a head injury between BOSS1 and BOSS2 (OR=3.07,
head trauma. Subjects were healthy academics who reported 95%CI=1.23, 7.64) were associated with an increased risk for
they had experienced a concussion, loss of consciousness decline in SDOIT score. In a multivariable model adjusting
or loss of memory from a head injury. In a questionnaire for age, sex and head injury, the number of carotid artery sites
study (N=3807), weight was significantly elevated and so with plaque was an independent predictor of decline in SDOIT
was preference for high fat foods controlling for age and score (OR=1.37, 95%CI=1.11, 1.68). Smoking and IMT were
not significant in models including plaque. These preliminary #P16 POSTER SESSION I: MULTIMODAL
findings suggest atherosclerosis may be a contributor to, or RECEPTION; CHEMOSENSATION AND DISEASE;
marker of, the decline in olfactory function seen with aging. OLFACTION PERIPHERY
Acknowledgements: The project described was supported
by R01AG021917 from the National Institute on Aging, Comparison of olfactory bulb neuronal phenotypes in
National Eye Institute, and National Institute on Deafness control and Parkinson’s Disease patients
and Other Communication Disorders. The content is solely the John W Cave1,2, Harriet Baker1,2
responsibility of the authors and does not necessarily reflect the 1
Burke Medical Research Institute White Plains, NY, USA,
official views of the National Institute on Aging or the National 2
Weill Cornell Medical College New York, NY, USA
Institutes of Health.
Olfactory dysfunction is found in nearly all sporadic cases
#P15 POSTER SESSION I: MULTIMODAL of Parkinson’s Disease and typically manifests years before
RECEPTION; CHEMOSENSATION AND DISEASE; motor symptoms are detected. Since the cellular mechanisms
OLFACTION PERIPHERY underlying this disruption of olfactory function are not
understood, we have started immunohistological studies to
Measures of smell function in youth with autism establish whether select neuronal populations degenerate
E. Leslie Cameron1, Richard L. Doty2, Shereen J. Cohen3, in the olfactory bulbs of Parkinson’s patients. We are using
Karen R. Dobkins3 olfactory bulbs from age- and sex-matched patients that were
pathologically confirmed to be either control or Parkinson’s
1
Department of Psychological Science, Carthage College Disease patients. These studies will examine the expression
Kenosha, WI, USA, 2Smell and Taste Center, Department of of tyrosine hydroxylase, calbindin and calretinin to assess
Otorhinolaryngology: Head and Neck Surgery Philadelphia, whether subsets of inhibitory interneurons are altered. To
PA, USA, 3Department of Psychology San Diego, CA, USA address whether there are changes in the glutamatergic
Autism spectrum disorders (ASD) are pervasive develop- projection neurons, we will analyze the expression of Tbx21
mental disorders characterized by deficits in social, com- since it is selectively expressed in the mitral/tufted cells. We
municative, and emotional behaviors. In addition to these will also address whether Parkinson’s Disease disrupts either
hallmarks, there is evidence for atypical sensory process- cholinergic or serotonergic centrifugal inputs to the olfactory
ing. In particular, there is the suggestion of atypicalities in bulb by analyzing the expression of choline acetyltransferase
chemosensation, tested with either questionnaires or direct and serotonin, respectively. Together, these analyses will
smell tests. Here we used both measures concomitantly. determine whether alterations in specific olfactory bulb
Sixteen youth with ASD (mean 15.3 yrs, 4 girls) and 16 typi- neuronal populations underlie, at least partially, the cellular
cally developing youth (mean 14.3 yrs, 7 girls) participated. mechanism responsible for olfactory dysfunction associated
Self-reported sensory processing was measured with the with Parkinson’s Disease, which will significant advance our
Adolescent/Adult Sensory Profile. This 60-item question- understanding how the disease progresses and potentially
naire includes items related to the five major senses and con- provide insight for future studies to develop novel therapeutic
ceptually arranged into four categories – “low registration” strategies. Acknowledgements: NIH DC008955
(i.e. low sensitivity to stimuli), “sensory sensitivity” (i.e. high
sensitivity to stimuli), “sensation seeking”, and “sensation #P17 POSTER SESSION I: MULTIMODAL
avoiding”. Smell function was measured with the pediatric RECEPTION; CHEMOSENSATION AND DISEASE;
Smell Wheel (Cameron & Doty, 2012). This scratch and sniff OLFACTION PERIPHERY
test measures the ability to identify 11 common odors using
a 4-alternative forced-choice procedure using pictures and Waist to hip ratios predict odor recognition memory
words to reduce cognitive load. Participants also rated the processing speed in carriers of the Apolipoprotein E
pleasantness of each odor. Youth with ASD scored signifi- e4 allele
cantly higher on “low registration”, “sensory sensitivity” and Melissa Cervantez1, Lisa Graves1, Amanda Green2, Charlie
“sensation avoidance”, and significantly lower on “sensation Morgan1, Claire Murphy1,2
seeking” (p<0.05). However the ASD and control groups did 1
San Diego State University San Diego, CA, USA, 2University
not differ in their ability to identify odors, nor were there
of California, San Diego La Jolla, CA, USA
differences in hedonic ratings of odors. More sensitive meas-
ures of olfactory function may be needed to detect differ- The ε4 allele of Apolipoprotein E (ApoE ε4) is currently the
ences in smell function in autism. Alternatively, self-reported strongest genetic risk factor associated with Alzheimer’s dis-
sensitivity to olfactory stimuli may reflect altered perception ease (AD). Studies have linked the ε4 allele with olfactory
of odors (e.g., heightened annoyance) rather than changes decline. Previous research has also found that waist to hip
in sensory sensitivity. Acknowledgements: Psi Chi Faculty ratios predict olfactory event-related potential (OERP) laten-
Advisor Research Grant cies among ε4+ individuals at the P3 cognitive component.
The current study aimed to measure OERPs among ε4+ effect of non- occupational exposure to Mn on olfactory
and ε4− individuals as they completed an odor recognition function. We therefore investigated peripheral and central
memory task and to determine whether the waist to hip ratio olfactory performance in a non-occupationally Mn-exposed
can predict the latency of the N1 sensory component of the population. Using the Sniffin’ sticks test battery, we compared
OERP waveform. OERPs were recorded from the FZ, CZ, the olfactory performance of two groups (n = 30/group) from
and PZ midline scalp electrode sites in 60 participants (30 F, a Mn mining district in central Mexico: an exposed group
30 M, mean= 46.88 years) with an equal number of ε4+ and living <1 km from a Mn processing plant, and a non-exposed
ε4− individuals. The odors were presented by a computer- control group living 50 kilometers from the closest source of
controlled olfactometer. Participants were instructed that exposure. Groups were matched for age, sex and schooling,
they were performing a memory task and completed three and none had ever worked in mining-related activities.
sessions: session 1 was an exposure trial for encoding, session Concentrations of Mn in hair (MnH) were measured as a
2 was a retrieval trial using odors, and session 3 was a retrieval biomarker of exposure. Exposed subjects had significantly
trial using odor labels. Using a bivariate correlation analysis, higher median MnH values (9.45 µg/g) than non-exposed
results indicated a significant positive correlation between subjects (1.04 µg/g). Furthermore, they were significantly out-
waist to hip ratio and N1 latency during odor retrieval hits for performed by the controls on all olfactory measures (threshold,
the FZ, CZ, and PZ electrode sites (r = .358, p = .006; r = .352, discrimination, and identification), indicating adverse effects
p = .007; r = .334, p = .010). When ApoE ε4 allele status was of Mn exposure on a range of olfactory functions, including
examined separately, the positive correlation remained signifi- those involving higher-order cognitive processes. These results
cant for ε4+ individuals (r = .439, p = .015; r = .423, p = .020; are in contrast to previous findings from our group showing
r = .425, p = .019) but not for ε4− individuals. The results adverse peripheral but not central effects on olfactory function
suggest that latency of the N1 OERP component during of big city air pollution, which mostly consists of toxicants
retrieval of an odor recognition memory task may be a useful known to affect the OE but with lower transsynaptic transport
measure for examining the negative effects of high waist to hip capacity compared to Mn. Conclusion: non-occupational
ratios in those genetically at risk for AD. Acknowledgements: exposure to air-born Mn can adversely affect both peripheral
Supported by NIH grant # DC002064-14 from the NIDCD and central olfactory function. Acknowledgements: MG
and A6004085-25 from the NIA. We thank Paul Gilbert for received a doctoral grant from CONACYT.
his statistical expertise and Derek Snyder, Jessica Bartholow,
Roberto Zamora, Ariana Stickel, Kyle Sigel, Jean-Loup
Bitterlin, Kristina Constant, and Sanae Okuzawa for helping #P19 POSTER SESSION I: MULTIMODAL
with data collection, entry and analysis. RECEPTION; CHEMOSENSATION AND DISEASE;
OLFACTION PERIPHERY
Odor perception and cerebral odor processing in adults
with autism spectrum condition.
RECEPTION; CHEMOSENSATION AND DISEASE;
OLFACTION PERIPHERY Luzie K. Koehler1, Cornelia Hummel1, Katja Albertowski2,
Veit Roeßner2, Thomas Hummel1
Adverse effect of non-occupational atmospheric exposure
to manganese on peripheral and central olfactory function
1
Smell & Taste Clinic, Department of Otorhinolaryngology,
University of Dresden Medical School Dresden, Germany,
Marco Guarneros1,2, Nahum Ortiz-Romo1, Mireya Alcaraz- 2
Department of Children and Youth Psychiatry, University of
Zubeldía3, Matthias Laska4, René Drucker-Colín1, Robyn
Dresden Medical School Dresden, Germany
Hudson5
Children and also adults with autism spectrum condition (ASC)
1
Instituto de Fisiología Celular, Universidad Nacional
are known to exhibit certain abnormalities in their sensory per-
Autonoma de Mexico (UNAM) Mexico City, Mexico,
ception. However, there are only few studies on the sense of smell
2
Posgrado en Ciencias Biológicas, UNAM Mexico City,
in autism. The aim of this study was to investigate whether there
Mexico, 3Instituto Nacional de Neurología y Neurocirugía
are differences in brain activation in response to odors between
Mexico City, Mexico, 4Department of Physics, Chemistry and
adults with autism spectrum condition and healthy controls.
Biology, Linköping University Linköping, Sweden, 5Instituto
To this end both structural (volume of the olfactory bulb) and
de Investigaciones Biomédicas, UNAM Mexico City, Mexico
functional measurements (fMRI) were performed. In addition
Manganese (Mn), although an essential trace element, is subjects underwent extensive psychophysical tests of olfactory
of growing concern as a toxic air pollutant. It is not only function („Sniffin’ Sticks”: olfactory detection thresholds, odor
readily transported from the olfactory epithelium (OE) to the identification). Twenty-two adults with ASC (high functioning
olfactory bulb but, unlike other metals, is also transported autism and Asperger’s syndrome) and 22 healthy controls were
transsynaptically to structures deep within the brain. However, examined. Results indicate a significantly impaired olfactory
to our knowledge no information is available on the possible threshold in ASC patients. In addition, olfactory identification
was found to be significantly worse in patients with ASC. In interaction between gastrointestinal functions and olfactory
terms of the size of the olfactory bulb no significant difference perception. Acknowledgements: ELAN-Application/FAU, in
between the two groups was found. With regard to fMRI, ASC part by Neurotrition Project, FAU Emerging Fields Initiative.
patients exhibited a higher level of overall activation in contrast
to control participants. Furthermore, patients with ASC also
showed significantly more activation in areas of the brain typi- #P21 POSTER SESSION I: MULTIMODAL
cally associated with odor processing, e.g. the orbitofrontal cor- RECEPTION; CHEMOSENSATION AND DISEASE;
tex and the limbic system. To summarize, even though impaired OLFACTION PERIPHERY
thresholds and results for odor identification indicate a lower Effects of Parkinson’s Disease on the Global Field Power of
level of olfactory function, a higher level of brain activation Olfactory Event-Related Potentials
during odor processing was found.
Allen Osman1, Ellen Carson1, Ian Pawasarat1, Fidias E. Leon-
Sarmiento1, Jonathan Silas2, Richard L. Doty1
#P20 POSTER SESSION I: MULTIMODAL 1
Smell and Taste Center, University of Pennsylvania
RECEPTION; CHEMOSENSATION AND DISEASE; Philadelphia, PA, USA, 2Department of Psychology,
OLFACTION PERIPHERY Roehampton University London, United Kingdom
Subjective and objective olfactory abnormities in Crohn`s Aim: Measurement of event-related brain potentials in
disease response to olfactory stimulation (OERPs) most often involves
the amplitude and/or latency of specific components. These
Marie Nitsch1, Yurdaguel Zopf2, Marion Clepce1, Dieter
components however can be difficult to discern in OERPs from
Schwab3, Kornhuber Johannes1, Thuerauf Norbert1
a single individual, especially one who is hyposmic. To circum-
1
Department of Psychiatry, Friedrich-Alexander-University vent this problem, we explored the use of an alternative meas-
Erlangen Nuremberg 91054 Erlangen, Germany, ure that reflects the overall strength of an OERP: Its Global
2
Department of Medicine 1, Friedrich-Alexander- Field Power (GFP). Methods: The subjects were 20 patients
University Erlangen Nuremberg 91054 Erlangen, Germany, with early-stage Parkinson’s disease (PD) and 20 matched
3
Department of Medicine II, Martha-Maria Hospital 90491 healthy controls. Stimuli consisted of 200 msec. presentations
Nuremberg, Germany of hydrogen sulfide delivered via a continuous-flow olfactom-
The pathogenesis of Crohn`s disease (CD) is still unknown. eter. OERPs were recorded at 40 electrode sites across the scalp
An involvement of the olfactory system in CD appears and combined to form a single GFP trace based on their stand-
possible due to immunregulatory and environmental aspects. ard deviation at each time-point. Results: It was possible to
To date no study has systematically assessed the olfactory obtain GFP measures for almost all subjects. The magnitude
function in CD patients. Therefore, we investigated subjective of change in GFP following odorant presentation, i.e. the over-
and objective olfactory function in CD patients. 31 CD all strength of the OERP response, was smaller for PD than
patients in active disease, 27 CD patients in inactive disease control subjects. Conclusions: While previous work has found
and a control sample of 35 age and sex matched healthy effects of PD on OERP latency, the present study shows that
volunteers were included in the study. Subjective testing was there is an effect also on amplitude. More generally, this study
applied using the Sniffin Sticks Test. For olfactory testing demonstrates that a fully objective OERP measure is sensitive
olfactory event-related potentials (OERPs) were obtained by to olfactory function and can be obtained readily from single
a four channel olfactometer using phenyl ethyl alcohol (PEA) individuals. As such, OERP GFP may be of use in the clinic and
and hydrogen sulfide (H2S). Chemosomatosensory event- well suited for standard statistical analyses. Acknowledgements:
related potentials (CSSERPs) were obtained using carbon Supported by USAMRAA W81XWH-09-1-0467.
dioxid (CO2) as a control stimulus. The analysis of variance
revealed significant higher hedonic ratings for pleasant #P22 POSTER SESSION I: MULTIMODAL
odorants in CD patients compared to healthy controls. RECEPTION; CHEMOSENSATION AND DISEASE;
Furthermore a trend to a lower threshold in CD patients in OLFACTION PERIPHERY
inactive disease occurred. In the objective olfactory testing
CD patients showed lower base-to peak-amplitudes (N1) Sensory Modality Influences Episodic Metamemory
and lower peak-to-peak-amplitudes (P1–N1). Additionally Accuracy in Healthy Aging and Alzheimer’s Disease
the OERPs showed significant shorter N1- and P2-latencies Jacquelyn Szajer1, Claire Murphy1,2
following the unpleasant odorant (H2S) in the right nostril 1
San Diego State University San Diego, CA, USA, 2University
for inactive disease compared to healthy controls. Our results
of California San Diego San Diego, CA, USA
demonstrate specific abnormities of olfactory perception in
CD patients, which could be interpreted as a natural regulation Episodic memory is commonly known to decline in both
against malnutrition. This points out the significance of the healthy aging and Alzheimer’s disease (AD), however, these
declines are heterogeneous. One factor shown to influence a cure. It has been shown that the pathology of AD (amy-
declines in episodic memory is metamemory, which is loid beta plaques (Aβ) and neurofibrillary tangles (NFT)) are
known to play an essential role in the strategies used for the first found in areas involved in olfaction. Decreased sense of
encoding and retrieval of information, as well as the control smell is seen in the earliest stages of AD and in Mild Cognitive
of memory output (Pannu & Kaszniak, 2005). Prior research Impaired (MCI) patients. In this study we used olfactory func-
has shown that memory-based olfactory tests are useful in tional Magnetic Resonance Imaging (fMRI) and volumetric
gauging cognitive decline in aging and AD (Murphy, Nordin, MRI to examine the relationship between the functional
& Jinich, 1999), and the current study aimed to examine the deficit and the pathological changes (atrophy) in the primary
utility of these tests in assessing declines in metamemory olfactory cortex (POC) and in the hippocampus in 23 cogni-
accuracy. Using a sample of 109 older adults with mild to tively normal controls (NC), 19 MCI, and 15 AD subjects.
moderate Alzheimer’s disease and 97 healthy controls, the The volumetric data shows that the volume of the POC is sig-
effect of diagnosis and sensory modality on metamemory nificantly different between the three groups (p <0.05); specifi-
accuracy (operationally defined as confidence in the accuracy cally the NC had a larger POC than the AD group. The fMRI
of responses at retrieval) was analyzed using an episodic data (p <0.05) also shows significant differences between the
recognition memory task comprised of both olfactory three groups (p <0.05); specifically the NC have significantly
and visual stimuli. Results indicated that both diagnosis more activated voxels in the POC than the AD group. The vol-
and modality had main effects on confidence accuracy (p ume and activation in the hippocampus were also measured
<.05). Overall, healthy older adults reported more accurate to see how the POC compared. The volume of the hippocam-
confidence levels than those with AD, and confidence levels pus and POC are positively correlated as well as the number
for the odor modality were less accurate than for the visual of activated voxels within the two regions of interest. These
modalities. Additionally, the interaction between diagnosis data show that the POC is involved in early AD and contrib-
and sensory modality was significant (p <05); controls utes to the olfactory deficits observed in early AD patients.
reported significantly more accurate confidence levels in Acknowledgements: Leader Family Foundation, NIH
correct responses than those with AD in the visual modalities,
but not the olfactory modality, indicating healthy older
adults are no better at judging odor memory performance #P24 POSTER SESSION I:MULTIMODAL
than those with AD, despite marked AD-related declines in RECEPTION; CHEMOSENSATION AND DISEASE;
olfaction. Applications and implications will be discussed. OLFACTION PERIPHERY
Acknowledgements: Supported by NIH grant # AG04085- The Effect of Abeta Accumulation on Odor Processing in
25 (C. Murphy) with a Diversity Supplement (J. Szajer), Anterior Piriform Cortex
and grant #AG005131-28 to the UCSD ADRC from the
National Institute on Aging. WenJin Xu1,3, Stephanie Lauer1,2, Chelsea Schoen1, Mirielle
Lopez-Guzman1, Efrat Levy2,4, Ralph Nixon2, Donald
A Wilson1,3
Emotional Brain Institute, Nathan Kline Institute
RECEPTION; CHEMOSENSATION AND DISEASE; Orangeburg, NY, USA, 2Center for Dementia Research,
OLFACTION PERIPHERY Nathan Kline Institute Orangeburg, NY, USA, 3Department
Primary Olfactory Cortex is affected in Alzheimer’s of Child and Adolescent Psychiatry, NYU Langone Medical
Disease and Mild Cognitively Impaired Patients: Center New York, NY, USA, 4Department of Biochemistry
A neuroimaging study and Molecular Pharmocology, NYU Langone Medical Center
New York, NY, USA
Megha Vasavada1, Jianli Wang1, Xiaoyu Sun1, Christopher
Weitekamp1, Paul Eslinger2, Prasanna Karunanayaka1, Sarah Alzheimer’s disease (AD) is a neurodegenerative disorder that
Ryan1, Qing Yang1,3 is the main cause of dementia in the elderly today. Previous
studies have implicated the accumulation of amyloid beta
1
Radiology, Penn State College of Medicine Hershey, PA,
(Ab) in the brain as the hallmark pathology seen in AD.
USA, 2Neurology, Penn State College of Medicine Hershey,
Patients suffering from AD often report problems with their
PA, USA, 3Neurosurgery, Penn State College of Medicine
sense of smell and indeed, cortical areas of the olfactory
Hershey, PA, USA
system are sites of early Ab deposition. Despite this temporal
Alzheimer’s Disease is a neurodegenerative disorder affect- prominence, olfactory degeneration in AD remains under-
ing 5.4 million Americans and it is the 6th leading cause of explored. Here, we conducted a cross-sectional study
death. Diagnosis of the disease is made when the pathol- using single-unit, ensemble and local field potential (LFP)
ogy has progressed to the neocortex and the effectiveness recordings to assess odor-evoked responses in the anterior
of drug intervention is unlikely. Therefore, early diagnosis is piriform cortex of anesthetized Tg2576 transgenic mice at 3,
key in understanding the disease progression and unlocking 6 and 12 months of age in order to investigate the effects of
Ab accumulation on odor processing. Stimuli included both NO errors was significantly higher in AD (27%) than PD (8%)
monomolecular odorants and complex odor mixtures. The or HC (5%) (p<0.0001). Odor anomia was also significantly
mixtures allow assessment of pattern completion processing worse in AD (15%) than PD (8%) and both were significantly
and are treated similarly in mice (shown here) as in rats (Barnes worse than HC (3%). While both PD and AD patients were
et al., Nat. Neurosci. 2008). Results suggest that, as previously impaired at NO identification, those with PD were more often
reported, Tg2576 were impaired in an odor habituation task as ansomic and those with AD were more likely to have an odor
compared to age-matched wild-type animals, Wesson et al. (J. agnosia and anomia. Acknowledgements: This work is sup-
Neurosci. 2010) and showed elevated aPCX LFP oscillations ported in part by the NIH/NCRR Clinical and Translational
(Wesson et al., J. Neurosci. 2011).. Preliminary single-unit Science Award to the University of Florida UL1 RR029890
and ensemble results suggest enhanced spontaneous firing and by the State of Florida Memory Disorders Program.
rates, abnormal odor receptive fields and impaired cortical
ensemble decorrelation of overlapping odor mixtures in
older Tg2576 mice compared to wildtypes. Ongoing analysis #P26 POSTER SESSION I: MULTIMODAL
includes investigation of the age of onset of these single- RECEPTION; CHEMOSENSATION AND DISEASE;
unit, ensemble and LFP changes and how they correlate with OLFACTION PERIPHERY
Ab deposition in olfactory structures and the emergence of
behavioral olfactory deficits. Acknowledgements: AG037693 Olfactory modulation of speech perception
Denise Chen1, Jennifer Chen2
Baylor College of Medicine, Neurology Department
1
Houston, TX, USA, 2Rice University, Psychology Department
RECEPTION; CHEMOSENSATION AND DISEASE; Houston, TX, USA
OLFACTION PERIPHERY
Speech perception can be modified by visual speech stimuli.
Differences in Odor Identification between Alzheimer’s and The canonical example is the McGurk Effect in which discord-
Parkinson’s Patients ant visual and auditory speech stimuli give rise to an illusory
Jennifer J Stamps1, Kenneth M Heilman2, Linda M auditory percept that is different from the percept generated
Bartoshuk1 in the presence of concordant visual and auditory stimuli.
Besides vision, tactile stimuli have also been shown to inte-
1
University of Florida Center for Smell and Taste Gainesville,
grate with auditory speech, suggesting that shared information
FL, USA, 2University of Florida Department of Neurology
across modalities underlies sensory integration in speech per-
Gainesville, FL, USA
ception. Here we investigate this by performing the McGurk
Whereas odor detection threshold is more impaired in patients paradigm in the presence of olfactory signals and examining
with Parkinson’s disease (PD) than Alzheimer’s disease (AD), the role of olfaction in modulating speech perception. Subjects
odor identification appears more impaired in AD than PD watched video clips of a mouth articulating a word while they
(Rahayel et al.’12). Failure to identify sensory stimuli can be simultaneously inhaled a subthreshold pleasant or unpleasant
induced by a sensory deficit (anosmia, failure to detect), a smell, and indicated the word they hear. The connotation of
perceptual recognition deficit (apperceptive agnosia, ability to the word can be positive or negative depending on the extent
detect the odor without the ability to correctly describe the of visual-auditory integration. We find that olfaction is able to
odor, and denies correct identification), or a failure of percept modulate the McGurk Effect, thereby showing the ubiquity of
to access lexical semantic networks (anomia, inability to name multisensory integration in speech perception.
with preserved ability to correctly describe the odor, select the
name from choices or agree when told the correct name). The
purpose of this study was to better understand the type of #P27 POSTER SESSION I: MULTIMODAL
olfactory identification error exhibited in 22 participants with RECEPTION; CHEMOSENSATION AND DISEASE;
AD, 23 with PD and 45 matched controls (HC) during an OLFACTION PERIPHERY
open- choice olfactory task (NO) with 23 food items. The per-
centages of correct odor identification of AD (39%) and PD The Dual Function of Basic Taste Stimuli: Signaling
(42%) participants were not different, but both were signifi- Nutrients in Smell and Taste
cantly lower than HC (83%, p<0.0001). In contrast, the aver- Jennifer Chen1, Denise Chen2
age NO intensity ratings of AD (29.3) were no different from
Psychology Department, Rice University Houston, TX,
1
HC (29.1) while PD average NO intensity ratings (14.5) were
USA, 2Neurology Department, Baylor College of Medicine
significantly lower than HC (p=0.001) and AD (p=0.005). In
Houston, TX, USA
addition, the average percentage of anosmic NO responses was
significantly higher in PD (41%) than in AD (21%, p=0.001) Olfaction and taste are often considered important in
and HC (8%, p<0.0001). The average percentage of agnosic guiding what we eat and drink. While much is studied about
the involvement of taste in sensing nutrients, little is known far, 74% were taste responsive, and 23% were odor respon-
about the role of olfaction in this process. Here we observe sive. There were no odor-responsive cells that were not
that the basic taste stimuli of monosodium glutamate also taste-responsive. When odorants are paired with taste
(MSG) and sucrose, commonly thought to be odorless, stimuli, 96% of the 31 cells recorded showed either sup-
generate distinctive olfactory sensations. In addition, pression or enhancement of taste responses. In addition,
subjects’ olfactory sensitivity to MSG and sucrose, unlike a small group of non-taste- or odorant- responsive cells
that to a nonfood smell, is modulated by their physiological (n = 8) responded when presented with a paired odorant-
states of hunger and satiety. Furthermore, at suprathreshold, tastant stimulus. These results suggest that multisensory
MSG and sucrose smells prolong the visual dominance processing occurs at the initial stages of gustatory pro-
of nutrient-congruent food in a binocular rivalry task; at cessing. Acknowledgements: Supported by NIDCD grant
subthreshold, MSG smell and taste summate and boost RO1DC006914 to PMD.
the visual perception of protein-rich food. These effects
occur despite the fact that subjects are not verbally aware
of the nature of the smells or their sensory differences. #P29 POSTER SESSION I: MULTIMODAL
Our findings offer the tantalizing evidence that basic taste RECEPTION; CHEMOSENSATION AND DISEASE;
stimuli serve the dual function of signaling nutrients in OLFACTION PERIPHERY
smell and taste, and point to the hitherto unsuspected role Enhancement of odor intensity and hedonics by taste: roles
of olfaction, alone or in combination with taste and vision, of nutritive taste and congruency
in monitoring homeostatic needs and guiding the search for
nutrient-rich food. Tomomi Fujimaru, Juyun Lim
Oregon State University Corvallis, OR, USA
#P28 POSTER SESSION I: MULTIMODAL We have previously shown that sucrose enhances the
RECEPTION; CHEMOSENSATION AND DISEASE; perceived intensities of congruent retronasal odors,
OLFACTION PERIPHERY whereas caffeine and citric acid do not. The present study
is designed to investigate whether saltiness and umami
Taste and Odor Convergence in the Nucleus of the Solitary can also enhance retronasal odors. In addition, given
Tract of Awake, Behaving Rats. our previous finding that taste-odor congruency plays an
Olga D Escanilla, Patricia M Di Lorenzo important role in retronasal odor referral to the mouth,
we tested the roles of congruency in the enhancement of
Binghamton University/Psychology Department
odor intensities and hedonics. Tomato and chicken odors
Binghamton, NY, USA
were presented alone or with NaCl, KCl, MSG, MPG,
Although many studies have shown the importance of or caffeine. Using a sip-and-spit procedure, Ss rated
gustatory and olfactory interactions in flavor percep- 1) the degree of liking/disliking of flavor on the LHS;
tion, very little is known about multisensory interaction 2) the intensities of saltiness, savoriness, bitterness and
in the initial stages of gustatory processing. Previously, it specific odor on the gLMS; and 3) the degree of taste-
has been shown that a subset of cells in the nucleus of odor congruency on a VAS. The result showed that both
the solitary tract (NTS; Van Buskirk & Erickson, Brain salty (NaCl) and umami (MSG, MPG) tastes significantly
Res.,135(2):287–303, 1977) and the parabrachial nucleus enhanced the perceived intensities of tomato and chicken
of the pons (PbN; Di Lorenzo & Garcia, Brain Res odors (Tukey test, p<.05), while other tastes failed to
Bull.,15(6):673–6, 1985) in rats respond to both taste and enhance the odor intensities. Similarly, NaCl, MSG, and,
olfactory stimuli. Here, we studied whether taste cells in to a lesser extent, MPG enhanced the degree of liking
the NTS of the awake, behaving animal could also respond for both odors, whereas KCl and caffeine significantly
to olfactory stimuli. Rats were surgically implanted with a decreased the odor liking (p<.05). The data also showed
microwire bundle into the NTS and allowed to recover. that the degrees of enhancement for both odor intensity
Rats were mildly water deprived and placed in an experi- and hedonics were significantly correlated with the degrees
mental chamber containing a lick spout for taste stimulus of taste-odor congruency (r=.95–.99, p<.01). Overall,
delivery and an odor port for olfactory stimulus delivery. our findings suggest that the presence of a nutritive taste
Tastants (0.1 M NaCl, 0.1 M sucrose, 0.01 M citric acid, (that signals the presence of a macronutrient) is required
0.0001 M quinine and artificial saliva) were delivered for for retronasal odor enhancement to occur and that taste-
5 consecutive licks interspersed with 5 licks of artificial odor congruency may further modulate the degree of odor
saliva rinse delivered on a variable ratio 5 schedule. All enhancement. The current data also support the notions
taste stimuli were presented both with and without an that salty and umami tastes can increase the palatability of
odorant in separate trials. Odorants were 1 Pa n-amyl ace- food odors and that the degree of taste-odor congruency
tate, 1 Pa acetic acid and air. Of the 31 cells recorded thus can predict the degree of flavor liking.
#P30 POSTER SESSION I:MULTIMODAL interaction that has received less attention than those with
RECEPTION; CHEMOSENSATION AND DISEASE; other taste sub-modalities (viz., sweet, sour, salty, and
OLFACTION PERIPHERY savory). These experiments used a computer-controlled
olfactometer-gustometer to simultaneously present taste
The primary qualities evoked by quinine, sucrose and solutions and odorized air to the mouth. One experiment
capsaicin associate with propylthiouracil bitterness, but not replicated a known effect, viz. enhancement of sweetness
TAS2R38 genotype by a fruity-smelling ester, using the new olfactometer-
John E Hayes1, Alissa L Allen1, Nadia K Byrnes1, Emma L gustometer. A second experiment examined the effect of
Feeney1, John E McGeary2 a “burnt” odor on bitter intensity. Subjects included 12
1
Department of Food Science University Park, PA, USA, healthy men and women, aged 18 to 65. Taste solutions
2
Providence VA Medical Center Providence, RI, USA included five concentrations each of sucrose (sweet) or
Genetic variability in the ability to taste thiourea compounds sucrose octaacetate (SOA, bitter). Odorants included four
has been studied for over 80 years. In the last 2 decades, the concentrations each of ethyl hexanoate (sweet, pineapple)
perceived intensity of concentrated propylthiouracil (PROP or isovaleraldehyde (burnt meat). Subjects held solutions
bitterness) has become a common measure of individual dif- in the mouth for several seconds, and rated the strength
ferences in taste response and taste acuity, as PROP associ- of both taste and odor sensations before expectorating.
ates with greater intensity responses from a broad range of Subjects rated intensity using the general labeled magnitude
stimuli, including non-bitter tastants, irritants and even ret- scale (gLMS). Ratings of sweetness and bitterness increased
ronasal aromas. However, early work indicated the putative with the concentrations of sucrose and SOA, respectively.
receptor was specific for compounds containing the N–C=S This expected dose-response relationship supported the
moiety, and much but not all of the variation in PROP bit- validity of the ratings. Further, consistent with published
terness can be explained by polymorphisms in the TAS2R38 results, ethyl hexanoate significantly enhanced the rated
receptor gene. Still, it is hard to envision how a N–C=S spe- sweetness of sucrose solutions. Thus, the current method of
cific receptor is related to overall orosensory response. Here, automated testing proved sensitive to known enhancement
we report data for 100+ individuals tested in our laboratory effects. Finally, isovaleraldehyde significantly enhanced the
who had been genotyped for TAS2R38 and phenotyped for bitterness of SOA solutions, demonstrating that odors can
PROP. These participants also reported the intensity of qui- enhance bitter taste. In conclusion, the method performed
nine, capsaicin, and sucrose on a general Labeled Magnitude well, and modulation of bitter taste by retronasal odors
Scale. Our data replicate earlier reports associating PROP seems like a promising area for further investigation.
bitterness with the primary qualities of sucrose, quinine and
capsaicin. However, we also observed that the correlations #P32 POSTER SESSION I: MULTIMODAL
between the intensity of sucrose, quinine and capsaicin were
much stronger with each other than with PROP. As expected,
TAS2R38 genotype did not associate with the intensity of
OLFACTION PERIPHERY
these sensations. When individuals were split by genotype, the Understanding Valence: the Neurobiology of Appetitive
strength of the PROP-capsaicin and PROP-sucrose relation- and Aversive Odor-Taste Learning in Rats
ships increased substantially within the groups of homozy- SiWei Luo, Matthew Einhorn, Kim M. Gruver, Nana Fujiwara,
gous individuals. Collectively, this suggests PROP bitterness Thomas A. Cleland
is a confounded phenotype that captures both genetic vari-
ation specific to N–C=S compounds and overall orosensory Cornell University/Psychology Ithaca, NY, USA
response. Acknowledgements: Supported by funds from the Appetitive and aversive conditioning both exert effects
Pennsylvania State University and NIH grant DC0010904. on perceptual learning, but are qualitatively distinct; for
example, they evoke activity in different brain regions.
#P31 POSTER SESSION I:MULTIMODAL Here, we present an olfactory conditioning paradigm capa-
RECEPTION; CHEMOSENSATION AND DISEASE; ble of evoking both forms of conditioning with minimal
OLFACTION PERIPHERY procedural differences, enabling conditioning effects to be
directly compared using identical behavioral and physio-
The Effect of Retronasal Odor on Ratings of Sweetness and
logical analyses. Male Long- Evans rats (Rattus norvegicus)
Bitterness
served as subjects for this study. Using implanted intra-oral
Tomoyuki Isogai1,2, Paul Wise2 cannulae, appetitive or aversive tastants (0.20% saccharin
1
Milk science Institute, Megmilk Snow Brand Co.,Ltd. or 0.02 M quinine) were directly infused into rats’ mouths
Kawagoe, Japan, 2Monell Chemical Senses Center and paired (or backward-paired) with an odor conditioned
Philadelphia, PA, USA stimulus over 3 days such that training procedures differed
We report the first in a planned series of experiments only in the valence of the tastant. Rats displayed appro-
on how odors affect rated bitterness, a potential flavor priate anticipatory behaviors (rapid mouth movements,
tongue protrusions, gaping) in response to odors predict- exhibited the lack of Trpm5 expression in the MOE. Taken
ing tastant infusions, indicating that rats learned the asso- together, these data demonstrate that Pou2f3 expression
ciation between odors and tastants. Aversively-conditioned is associated with the expression of Trpm5 in multiple
rats also appeared to exhibit broader generalization to sim- types of chemosensory cells, and suggest that Pou2f3 is a
ilar odorants than did the appetitive and control groups. master regulator of differentiation of Trpm5-expressing
Immediate- early gene (Egr-1 and c-Fos) expression was chemosensory cells in digestive and respiratory epithelia.
measured in olfaction-, valence-, and conditioning-asso- 1
Nat. Neurosci. 14, 685–687 (2011)
ciated brain regions. IEG expression was higher in the
main olfactory bulb, anterior olfactory nucleus, piriform
cortex, and orbitofrontal cortex in aversively-conditioned #P34 POSTER SESSION I:MULTIMODAL
rats compared with appetitive and control groups. These RECEPTION; CHEMOSENSATION AND DISEASE;
results provide a foundation for studies of learning and OLFACTION PERIPHERY
plasticity in which appetitive and aversive associations
can be mechanistically compared in a common neural net- Aronia Berry Juice Sensory Analysis by Harvest Time and
work. Acknowledgements: Liu Memorial Award, Sigma Xi Oral Sensory Phenotype
Research Grant, Cornell University SAGE Fellowship Jeeha Park1, Shristi Rawal1, Mark H Brand2, Shelley
Durocher2, Mastaneh Sharafi1, Valerie B Duffy1
University of Connecticut/Allied Health Sciences Storrs, CT,
1
#P33 POSTER SESSION I:MULTIMODAL USA, 2University of Connecticut/Plant Science and Landscape
RECEPTION; CHEMOSENSATION AND DISEASE; Architecture Storrs, CT, USA
OLFACTION PERIPHERY
Aronia berries (chokeberries) have very high levels of
Pou2f3/Skn-1a is involved in the differentiation of multiple health- promoting antioxidants yet can cause a “choking”
types of Trpm5-expressing chemosensory cells sensation due to bitterness and astringency. We aimed to
Makoto Ohmoto1, Tatsuya Yamaguchi2, Keiko Abe3, describe oral sensations and palatability of aronia juice,
Alexander A. Bachmanov1, Junji Hirota2, Ichiro Matsumoto1 including variation by harvest time and oral sensory
phenotype. Ripe aronia berries were harvested at 7 time
1
Monell Chemical Senses Center Philadelphia, PA,
points and juiced for oral sampling by 50 adults who were
USA, 2Tokyo Institute of Technology, Department of
phenotyped for chemosensory function by the NHANES
Bioengineering Yokohama, Japan, 3The University of Tokyo,
protocol, olfactometer, and propylthiouracil (PROP) bit-
Department of Applied Biological Chemistry Tokyo, Japan
terness. The adults reported quality intensities of proto-
A homeodomain transcription factor Pou2f3 (also known as typical tastes, oral chemesthetic compounds, foods, berry
Skn-1a) is specifically expressed in sweet, umami, and bitter juice and non-oral stimuli, which served as sensory stand-
taste receptor cells in taste buds, and is necessary for their ards. The juice qualities correlated with alum astringency,
functional differentiation1. Recent studies have shown that quinine bitterness, and citric acid sourness but, unlike
taste receptors and/or signaling molecules indispensable for apple or grapefruit juices, did not correlate with sucrose
sweet, umami, and/or bitter taste are expressed not only in sweetness. The average berry juice response was weakly
gustatory tissues but also in the intestinal and respiratory dislike (range—v. strongly dislike to above moderately
epithelia. In this study, we examined the expression of like). Astringency was the strongest sensation, yet sweet-
Pou2f3 in the extra-oral epithelial chemosensory cells and ness was the primary driver of liking in multiple regres-
the impact of Pou2f3 knockout on the cells. In the intestinal sion analysis. Those who liked the juice reported a greater
epithelium, the expression of Pou2f3 was observed in balance between astringency and either sourness or sweet-
the tuft/brush cells that express Plcb2 and Trpm5 and ness. The juices were more sweet and less sour/astringent
participate in opioid secretion by chemosensing. Pou2f3- in the middle versus first harvest time, corresponding to
deficient mice lacked the expression of Plcb2, Trpm5, and greater acceptance. Classifying by PROP relative to qui-
other tuft/brush cell marker genes, but they still expressed nine bitterness identified adults who differed in intensities
characteristric genes of enteroendocrine cells. In the of prototypical oral stimuli but not in intensities of odors.
respiratory epithelium of the nasal cavity, the expression Those who perceived more PROP relative to quinine bit-
of Pou2f3 was observed in the solitary chemosensory cells terness also reported the juice as less sour, less balanced
(SCCs) that express Tas1r3, Tas2rs, Gnat3, Plcb2, and between astringency and sourness, and more disliked than
Trpm5. The expression of all these genes was lost in the those who perceived high bitterness from PROP and qui-
Pou2f3-defficient mice. We also found Pou2f3 expression nine. Further study aims to identify ways to maximize the
in a subset of microvillus cells in the main olfactory palatability of aronia juice, individualized to taste pheno-
epithelium (MOE) where Trpm5 but not Plcb2, Ggust, type. Acknowledgements: USDA/Hatch and USDA NE
or taste receptors were expressed. Pou2f3-deficient mice SARE
#P35 POSTER SESSION I: MULTIMODAL Department of Pharmacological and Physiological Science

RECEPTION; CHEMOSENSATION AND DISEASE; St. Louis University School of Medicine St. Louis, MO, USA
OLFACTION PERIPHERY Taste sensitive neurons throughout the gustatory neuraxis
Temperature of served water can influence sensory respond to oral somatosensory stimuli, like temperature.
perception and acceptance of subsequent food What is more, human psychophysical studies show that uni-
modal oral thermal stimuli can elicit taste perceptions, and
Han-Seok Seo1, Pauline Mony1,2, Tonya Tokar1, Peggy Pang1, increasing the temperature of a sucrose solution increases
Alexandra Fiegel1, Jean-François Meullenet1 its perceived “sweetness”. These data suggest that tempera-
1
University of Arkansas/Food Science Fayetteville, AR, ture may be modulating the neural signal for taste intensity.
USA, 2French National School of Agricultural Science and However, data on this topic are scarce. To investigate the
Engineering in Toulouse Castanet Tolosan Cedex, France potential influence of temperature on neural activity for taste
intensity we made extracellular recordings from single neu-
The cross-cultural difference in meal pattern exists in the
rons in the rostral nucleus of the solitary tract of anesthetized
typical temperature of water served with meals. For example,
C57BL/6J mice during oral application of temperature-and
North American people, as a whole, are used to drinking
concentration-varied tastants. Stimuli included purified
iced water/ beverages, while Asian or European people
water and a concentration series of sucrose (in M): 0.05, 0.1,
show a preference for hot water/tea or room temperature
0.17, 0.31, and 0.56 presented “whole mouth” at 17, 22, 30,
water, respectively. It has been proven that food perception
and 37º C. Preliminary analyses of 35 neurons show that
and acceptance are affected by oral temperature, as well as
warming sucrose to 37º C significantly reduced the response
by serving temperature of food. Based on the fact that the
onset latency compared to room temperature sucrose for all
iced or hot water served with meals can modulate the oral
concentrations tested (ps <0.01). Multivariate analysis shows
temperature, the present study aimed to determine if the
that temperature can substantially change stimulus-evoked
temperature of served water can modulate the sensory
across-neuron patterns of response in a way that differs from
perception of foods subsequently consumed. Following a
the influence of concentration. These analyses suggest that
mouth rinse with water served at 4, 20, and 50 °C for 5 s,
temperature is modulating taste responses in a unique way
two types of food: dark chocolate or cheddar cheese were
compared to concentration, suggesting temperature may
evaluated in terms of sensory intensity and overall liking.
be a parameter of the taste code. Acknowledgements: NIH
For the dark chocolate, the intensity ratings for sweetness,
DC011579 (C.H.L.)
chocolate flavor, and creaminess were significantly lower
when following water at 4 °C than when following water at
either 20 or 50 °C. However, the effect of water temperature #P37 POSTER SESSION I:MULTIMODAL
on sensory perception was not observed with cheddar RECEPTION; CHEMOSENSATION AND DISEASE;
cheese. In addition, the overall liking for the dark chocolate OLFACTION PERIPHERY
was significantly lower when following water at 4 °C than
when following water at either 20 or 50 °C. In conclusion, Olfactory modulation of visual temporal processing
the current study demonstrates new empirical evidence Bin Zhou, Guo Feng, Wen Zhou
that the consumption of iced water can decrease perceived
intensities of sweetness, chocolate flavor, and creaminess for Key Laboratory of Mental Health, Institute of Psychology,
Chinese Academy of Sciences Beijing, China
subsequently consumed chocolate. Our findings suggest a
possibility that the North American frequent consumption Perception is thought to consist of fast ‘snapshots’ of the
of iced water/soda may reduce their sensitivity to sweet world on a need-to-know basis. We ask if additional infor-
tasting stimuli, thereby leading to the preference for more mation from the olfactory channel, which naturally conveys
highly sweetened foods. Acknowledgements: This research object identities, influence the temporal encodings in the vis-
was supported by start-up funding from the University of ual system. Using a two-alternative forced choice method, we
Arkansas Division of Agriculture to HS SEO. find that a smell lengthens the subjective duration of a sensory
congruent visual object and enhances its visibility at frequen-
cies near critical flicker fusion (CFF) in a manner independent
#P36 POSTER SESSION I:MULTIMODAL of top-down cognitive control. In the latter case, the behavioral
RECEPTION; CHEMOSENSATION AND DISEASE; advantage is accompanied by increased power of neural oscil-
OLFACTION PERIPHERY latory responses over the occipital-temporal region around the
frequency of the flickering visual object. These results indi-
Stimulus temperature and concentration differentially
cate that olfaction modulates visual temporal processing at
influence the gustatory neural code for sucrose in the
the object representation level, and provide new insights into
mouse brain stem
the neural timing of multisensory events. Acknowledgements:
David M. Wilson, Christian H. Lemon National Basic Research Program of China (2011CB711000),
National Natural Science Foundation of China (31070906), Ishmail Abdus-Saboor, Benjamin Shykind
National Natural Science Foundation of China (31100735), Weill Cornell Medical College Qatar/ Department of Cell
Knowledge Innovation Program of the Chinese Academy of and Developmental Biology Doha, Qatar
Sciences (KSCX2-EW-BR-4 & KSCX2-YW-R-250)
Olfactory receptors (ORs) number more than 1,000 and
comprise the largest gene family in the mammalian genome.
#P38 POSTER SESSION I:MULTIMODAL ORs reside in heterochromatin and selection of one OR and
RECEPTION; CHEMOSENSATION AND DISEASE; from one allele is thought to occur stochastically. ORs are
OLFACTION PERIPHERY expressed both monogenically and monoallelically in olfac-
tory sensory neurons (OSNs) and the mechanism that con-
The Ice-Cream Effect: The Influence of Temperature on Taste
trols their regulation is largely unknown. Here we describe
Perception
results for mice with a ‘monoclonal’ nose that express one
Clare Wijngaarden1, Paul AM Smeets1,2, Sanne Boesveldt1 OR M71in 95% of all mature OSNs. M71 mice suppress
1
Wageningen University, Division of Human Nutrition expression of endogenous ORs, and expression of the sup-
Wageningen, Netherlands, 2UMC Utrecht, Image Sciences pressed ORs is shifted to the immature layer of the olfac-
Institute Utrecht, Netherlands tory epithelium. We show that the suppressed ORs were first
selected and then turned off by M71. When we introduced a
The Ice-cream Effect is a term coined to describe the second transgene into M71 mice that expressed another OR
anecdotally reported difference in perceived taste intensity in most mature OSNs, OSNs uncharacteristically expressed
of a product at different temperatures e.g. when using fruit both of the ORs. We hypothesize that unresolved OR com-
juice or syrup to make ice cubes, a more concentrated mix petition compromised the neuron’s ability to express only
is ideal for optimum taste when frozen compared with the one receptor. We further show that suppression of endog-
strength of a regular drink. The main objective of this study enous ORs by M71 is not reversible, and that M71 does not
was to investigate how temperature influences perceived need to be continuously expressed for endogenous ORs to
taste intensity and hedonic value for different solutions. 20 remain suppressed. In these experiments, we have engineered
females and 10 males (aged 18–34 years) evaluated solutions OSNs to express more than one OR in an OSN at a time,
of Sucrose (100g/L), Sucralose (0.254g/L), Na salt (8g/L), which is normally a low probability event. We have shown
Na/K salt mix (1:2 ratio, 11.128g/L) and milliQ water at that when this event arises, a secondary refinement pathway
approximately 1 (cold), 22 (ambient) and 50°C (hot) using is invoked that turns off one OR to maintain singular expres-
a sip and spit method. Solutions were presented as 10mL sion. We thus provide compelling evidence for a new para-
samples in randomised order and in duplicate. Participants digm of OR regulation: post-selection shut down, which we
rated the perceived intensity of four basic tastes (Sweet, hypothesize occurs through a yet-to-be uncovered competi-
Salt, Bitter, Sour), temperature and pleasantness on 10cm tive mechanism.
VAS for each sample. Overall there was no significant effect
of temperature on perceived taste intensity. Hot milliQ
samples were rated sweeter than ambient samples (p=0.038)
and hot Na/K salt samples as more pleasant than ambient RECEPTION; CHEMOSENSATION AND DISEASE;
or cold (p=0.024). Gender effects were found for perceived OLFACTION PERIPHERY
sweetness of Sucrose (Females rated > Males, p=0.016) and Odorant Receptor Dependent Spontaneous Firing Rates Do
pleasantness ratings of Na and Na/K salt samples (Males Not Predict Sensory-evoked Firing Rates in Mouse Olfactory
rated > Females, p=0.017 and 0.001). Gender*Temperature Sensory Neurons
interaction was found for pleasantness of Na/K salt samples
Timothy Connelly, Agnes Savigner, Minghong Ma
(p=0.025). Contrary to expectations, temperature did not have
a significant effect on perceived taste intensity. Nevertheless, University of Pennsylvania/Department of Neuroscience
it is suggested that sweet and salt tastes have greater hedonic Philadelphia, PA, USA
value at temperatures other than ambient and that factors Sensory systems need to tease out stimulation-evoked activity
such as changes in texture and oral exposure time should also against a background of spontaneous activity. In the olfactory
be considered as the basis for the ‘Ice cream effect’. system, the odor response profile of an olfactory sensory
neuron (OSN) is dependent on the type of odorant receptor
#P39 POSTER SESSION I:MULTIMODAL it expresses. OSNs also exhibit spontaneous activity, which
plays a role in establishing proper synaptic connections and
may also increase the sensitivity of the cells. However, where
OLFACTION PERIPHERY
the spontaneous activity originates and whether it informs
Evidence for a Cell Fate Refinement Mechanism in Olfactory sensory-evoked activity remain unclear. We addressed these
Sensory Neurons questions by examining patch-clamp recordings of genetically
labeled mouse OSNs with the defined odorant receptor M71 effect of longer-term adaptation on the peripheral olfactory
(n = 22), I7 (n = 21), SR1 (n = 11), mOR-EG (n = 24) or code. Acknowledgements: BBSRC UK grant BB/H009914/1
MOR23 (n = 16) in intact olfactory epithelia. We show that
OSNs expressing different odorant receptors had significantly
different rates of basal activity. Additionally, OSNs expressing #P42 POSTER SESSION I: MULTIMODAL
an inactive mutant I7 receptor completely lacked spontaneous RECEPTION; CHEMOSENSATION AND DISEASE;
activity (n = 34), despite being able to fire action potentials OLFACTION PERIPHERY
in response to current injection (n = 6). This finding strongly Characterization of the Iontransporter NKCC1 in the Field of
suggests that the spontaneous firing of an OSN originates Chemosensation
from the spontaneous activation of its G-protein coupled
odorant receptor. Lastly, we show that the spontaneous firing Claudia Haering, Janine Wäring
rates of selected OSN types do not correlate with the firing Hanns HattRuhr-University Bochum, Germany
rates evoked by a near-saturating odorant stimulus. This
The olfactory sense is mainly regulated through a cAMP-
study reveals that neither the basal activity nor the receptor
dependent signaling cascade which leads to a cation influx
type dictates the maximum odorant-evoked activity in OSNs,
and a chloride efflux through the neuronal membrane. This
which suggests that OSNs expressing the same receptor type
so called chloride boost during depolarization of olfactory
may send distinct information to the brain upon odorant
sensory neurons still remains unclear. In addition, how is the
stimulation. Acknowledgements: This work was supported by
R01 grants from NIDCD/NIH (DC006213 and DC011554). intracellular chloride concentration achieved in olfactory
neurons? Several publications demonstrated that the chloride
concentration is much higher in olfactory neurons, especially
#P41 POSTER SESSION I:MULTIMODAL in the knob, compared to surrounding cells and the mucus.
RECEPTION; CHEMOSENSATION AND DISEASE; NKCC1 is a candidate which fits the role of an ion transporter
OLFACTION PERIPHERY that causes the high chloride concentration inside olfactory
How plastic is the peripheral olfactory system of Drosophila neurons. NKCC1 is a 12 membrane spanning symporter of one
melanogaster larvae? sodium, one potassium and two chloride ions. The expression
of NKCC1 is confirmed in the olfactory epithelium of mice,
MA Grillet, R Petersen, C McCrohan, M Cobb
especially in olfactory neurons. However, the function of
University of Manchester Manchester, United Kingdom NKCC1 is controversially discussed in literature. In our project
we want to characterize NKCC1 knockout mice, thereby
During their larval stage, fruit flies are exposed to an odour-
addressing the question whether NKCC1 is involved in olfaction
rich environment, in which they must choose between toxic
and olfactory neurogenesis. On the one hand we are going to use
and edible substrates. For this they need an efficient olfac-
chloride imaging of acute slices of the olfactory epithelium of
tory system with the capacity for both short and long term
knockout and wild type mice. On the other hand morphological
plasticity. Drosophila larvae possess only 21 paired olfac-
studies and RNA fluorescence in situ hybridization (RNA
tory sensory neurons (OSN), most of which express a single
FISH) experiments will give us information about the role of
olfactory receptor (OR) type together with the co-receptor
NKCC1 in neurogenesis. Our first studies showed differences in
Orco. Information arising from each OSN is transmitted to
the morphology of the turbinates and the neuronal layer of the
a unique glomerulus in the antennal lobe and then to the
olfactory epithelium of NKCC1 knockout compared to wild
mushroom body via projection neurons. Combinatorial cod-
ing in the periphery allows larvae to detect and discriminate type mice leading to the question whether NKCC1 plays a role
in the continuous replacement of olfactory sensory neurons.
a large number of odours. Previous studies have character-
ised the odour-response profiles of 19 of the 21 OSNs and
found that a given OSN’s response to a given odour is often #P43 POSTER SESSION I: MULTIMODAL
highly variable (Hoare et al 2008, 2011). This raised the pos- RECEPTION; CHEMOSENSATION AND DISEASE;
sibility that the peripheral olfactory code exhibits plasticity
OLFACTION PERIPHERY
and that this plasticity may be directly involved in mediating
behavioural adaptation and learning. We are exploiting the Intrinsic electrophysiological property of Kenyon cells in
UAS-Gal4 system to create single OR lines in which only silkmoths
one identified OSN is functioning. We are studying the effect Shigeki Inoue1, Masashi Tabuchi2, Kei Nakatani1, Ryohei
of short and long-term adaptation on the response of indi- Kanzaki2
vidual OSN classes to a panel of odours using extracellular
electrophysiology and behavioural assays. Our preliminary
1
University of Tsukuba Tsukuba, Ibaraki, Japan, 2University
results show that short-term adaptation to specific odours of Tokyo Meguro, Tokyo, Japan
may induce a decrease or an increase in OSN responses Kenyon cells (KCs) are the component neurons of mushroom
depending on the odour used. We are currently exploring the bodies (MBs) of insect brain regions contributing to olfaction
and taste. In these contributions, KCs may have some impor- olfactory signaling. Through large-scale peptide microarrays
tant roles in olfactory information processing. However, the we could show numerous interactions between the 13 PDZ
intrinsic electrophysiological properties of silkmoth KCs domains of MUPP1 and different murine olfactory receptor
remain unknown. Here, we use whole-cell patch clamp record- C-termini of various subfamilys. Co-immunoprecipitations
ing to elucidate the functional parameters such as voltage-acti- validated the interactions between MUPP1 and the most
vated ionic currents of KCs in silkmoth MBs. KCs generated important signaling proteins in vivo and led to the assumption
action potentials in response to depolarizing current injections that an olfactory PDZome is organized by the scaffold MUPP1.
which were stepped in 20 pA between 0 and 100 pA, and appli- Furthermore co-immunoprecipitations in juvenile mice showed
cation of GABA-receptor blocker, picrotoxin (PTX) abolished no differences in binding properties in comparison to the adult
inhibitory synaptic inputs and depolarized resting potential. mice. To functionally show that olfactory signaling is PDZ-
By using voltage-clamp technique, we recorded membrane dependent we established patch clamping of olfactory sensory
currents including inward and outward voltage-activated cur- neurons in acute slices of transgenic mOR-EG mice. We also
rents. Pharmacological isolation of KC voltage-activated ionic created a small inhibitory peptide which disrupts the interaction
currents revealed that KCs express a range of voltage-activated between the mOR-EG receptor and MUPP1 and injected it
currents, including transient (Itransient; activated by voltage step through the patch pipette into the neuron. After uncoupling
pulses above −50 mV) and non-activating potassium (Isustained; the interaction between the mOR-EG receptor and MUPP1
activated by voltage step pulses above −30 mV), sodium (INa; the odor-evoked current amplitudes were strongly reduced and
activated by voltage step pulses above −50 to −40 mV) and the adaption was impaired, whereas a control peptide did not
calcium (ICa activated by voltage step pulse above −60 to −50 affect olfactory signaling. In conclusion, we confirmed that
mV) currents, and these potassium currents included calcium- an olfactory signalosome is mediated by MUPP1 in olfactory
activated components. Our results consisted with previous sensory neurons and showed that accurate olfactory signaling
research of cockroach (Demmer and Kloppenburg. 2009) and is a PDZ dependent mechanism.
provided the first electrophysiological characterization of KCs
in silkmoth MBs and suggested that the intrinsic properties
of KCs had common feature regardless of the insect species. #P45 POSTER SESSION I:MULTIMODAL
Our experiments represented an important step toward under- RECEPTION; CHEMOSENSATION AND DISEASE;
standing neural computation that underlies olfactory informa- OLFACTION PERIPHERY
tion processing in silkmoth. Expression of olfactory signaling molecules in the non-
chemosensory tissues
#P44 POSTER SESSION I: MULTIMODAL Nana Kang1, Hyoseon Kim1, Frank Margolis2, JaeHyung Koo1
RECEPTION; CHEMOSENSATION AND DISEASE; DGIST/Department of Brain Science Daegu, South Korea,
1
OLFACTION PERIPHERY University of Maryland/Department of Anatomy and

2
Scaffolding proteins in olfaction Neurobiology Baltimore, MD, USA
Fabian Jansen1, Sabrina Baumgart1, Willem Bintig2, Olfactory sense is mediated by specialized olfactory receptor
Benjamin Kalbe1, Christian Herrmann3, David Köster4, neurons (ORNs) in the nose. However, ectopic expressions
Sebastian Rasche1, Nils Metzler- Nolte4, Marc Spehr5, Hanns and functional roles of olfactory receptors (ORs) and olfac-
Hatt1, Eva Neuhaus2 tory signaling molecules (OMP, Gαolf, and AC3) still remain
to be elucidated. This study demonstrates the presence of
1
Department of Cell Physiology, Faculty for Biology and
olfactory signaling molecules in non-olfactory tissues by
Biotechnology, Ruhr-Universität Bochum Bochum, Germany,
systematically using RT-PCR, western blotting, immunohis-
2
Neuroscience Research Center, Cluster of Excellence
tochemistry, and a double-antibody immunoprecipitation/
NeuroCure, Charité Universitätsmedizin Berlin Berlin,
immunodetection procedure. Unexpectedly, the co-locali-
Germany, 3Department of Physical Chemistry I, Ruhr-
zation of OMP/AC3/Gαolf was confirmed in several tissues
Universität Bochum Bochum, Germany, 4Department of
while they were expressed on different cell types of the same
Inorganic Chemistry I, Ruhr-Universität Bochum Bochum,
organ in another non-chemosensory tissue. Additionally,
Germany, 5Department of Chemosensation, Institute for
gene expression of olfactory receptors (ORs) was observed
Biology II, RWTH- Aachen University Aachen, Germany
in non-olfactory tissues through RT-PCR. These results sug-
Mammalian olfactory signaling is dependent on various gest that olfactory receptors play an important role in tis-
proteins and fine-tuned through different processes. Most of sue-specific or common physiological functions of ectopic
the essential components are already known but the detailed expression in non-olfactory tissues. In the future, we need
organization of this complex system has yet to be understood. to define the physiological function of olfactory receptors
In our recent study we identified the multiple PDZ domain in non-chemosensory tissues. Acknowledgements: DGIST
protein MUPP1 as a potential scaffolding protein candidate for MIREBrain and Convergence Science Center (13-BD-0403)

Department of Neuroscience, Perelman School of Medicine
RECEPTION; CHEMOSENSATION AND DISEASE; at the University of Pennsylvania Philadelphia, PA, USA,
OLFACTION PERIPHERY
2
In vivo dynamic interactions between the methyl-CpG By converting environmental signals into intracellular
binding protein MeCP2 and chromatin under odor-evoked responses, cilia are critical for many biological processes,
activity including olfaction. Surprisingly, little is known about
what factors shape cilia morphology and how morphology
Wooje Lee, Qizhi Gong impacts function. We recently discovered that olfactory cilia
University of California at Davis/Cell Biology and Human vary considerably in length depending on the cell location
Anatomy Davis, CA, USA within the olfactory epithelium. Using specific markers
for a subset of olfactory sensory neurons (OSNs) from
MeCP2 was identified as a methyl-CpG binding protein
C57BL/6 mice (3–6 weeks, n = 19 animals), we found that
and capable of recruiting co-repressor complexes to pro-
cilia length increases from ~1 µm in the posterior nasal
moters to suppress gene expression. MeCP2 is abundant
septum to ~20 µm in the anterior septum, with the longest
in neurons. Mutations in MECP2 cause Rett syndrome, a
cilia (up to 50 µm) typically found in the dorsal recess. We
neurodevelopmental disorder. Recent studies suggest that
then built a 3D computational fluid dynamics model based
Mecp2 has multiple functions including transcriptional
on the mouse nasal cavity and demonstrated that cilia length
repression/ activation and structural compaction of chro-
is positively correlated with sensory inputs, particularly
matin. Dynamic interaction between MeCP2 and chro-
odorant absorption. To determine whether sensory inputs
matin is not well understood. The complexity of MeCP2
themselves account for the cilia length pattern, we performed
function among different neuronal populations and a dif-
unilateral naris closure on newborn mice (n = 6 animals) and
ferent methylation status in in vitro culture system have
immunostained olfactory cilia four weeks later. Remarkably,
made it challenging to understand MeCP2 binding profile
cilia length was increased in the open (overstimulated) nostril.
and dynamics under neuronal activity in vivo. Olfactory
We further found that cilia length modified OSN function,
epithelium provides an ideal in vivo model in its ubiquitous
as OSNs expressing a defined odorant receptor were more
neuronal population and accessibility for neuronal activity
sensitive to odorant stimulation when they had longer cilia
manipulation. In this study, we sought to identify MeCP2
(n = 7) as opposed to shorter cilia (n = 8). Together, these
binding profiles to different regions of the chromosome and
results suggest that sensory activity may shape olfactory
changes under odor-evoked activity. Chromatin immuno-
cilia length and, consequently, OSN function. This discovery
precipitation following high throughput sequencing shows
offers novel insight into the organization and function of
MeCP2 binds to not only methylated CpG island but also
OSNs and into cilia biology. Acknowledgements: Supported
intergenic and intronic regions and sparsely methylated
by NIDCD grants R01DC011554 and R01DC006213.
promoters. Genome-wide profiling for MeCP2 binding in
vivo clearly shows two distinct distributions of MeCP2,
one concentrated at regulatory regions and the other along #P48 POSTER SESSION I: MULTIMODAL
the entire genes locus. Odor-evoked activity results in sig- RECEPTION; CHEMOSENSATION AND DISEASE;
nificant changes in MeCP2 affinity to selected gene loci. OLFACTION PERIPHERY
Comparing methylation state and MeCP2 binding profiles
revealed that odor-evoked activity alters MeCP2 affinity SUMOylation regulates the ciliary localization of olfactory
to chromatin in a DNA methylation independent manner. signaling proteins
Our results reveal the complexity of MeCP2 and chroma- Jeremy C McIntyre, Jeffrey R Martens
tin interaction. We hypothesize that Mecp2 regulates activ-
University of Michigan, Department of Pharmacology Ann
ity-dependent gene regulations via changing its binding
Arbor, MI, USA
affinity to the entire gene locus. Acknowledgements: NIH
DC11346 In olfactory sensory neurons (OSNs), the protein compo-
nents for odor detection are highly enriched in cilia, however
the precise mechanisms for this localization remain poorly
#P47 POSTER SESSION I: MULTIMODAL defined. The mechanisms for selective cilia entry may be anal-
RECEPTION; CHEMOSENSATION AND DISEASE; ogous to nuclear import utilizing importin ß2, a Ran gradient
OLFACTION PERIPHERY and nucleoporins. A unique post-translational modification
process involved in nuclear-cytosolic transport is the revers-
Sensory inputs modulate olfactory cilia morphology and
ible conjugation of Small Ubiquitin-like Modifier (SUMO)
function in the mammalian nose
proteins or SUMOylation. Bioinformatic examination
Rosemary Lewis1, Huikai Tian1, Jiwei He1, Jianbo Jiang2, reveals that both adenylate cyclase 3 (AC3) and the calcium-
Timothy Connelly1, Kai Zhao2, Minghong Ma1 activated chloride channel, annoctamin2 (ANO2) harbor
conserved SUMOylation motifs. Therefore, we hypothesized To investigate PI signaling in OSNs, we will use adenoviral
that SUMOylation regulates ciliary localization of AC3 and vectors carrying two different fluorescently tagged proteins,
ANO2. Coexpression of SENP2, a SUMO protease, with the pleckstrin homology (PH) domains of phospholipase
either AC3:GFP or ANO2:GFP in MDCKII cells prevented C (PLC) and the general receptor of phosphoinositides
their normal ciliary localization. Site directed mutagenesis of (GRP1), to monitor PI activity in the murine olfactory
the predicted SUMOylation sites also blocked ciliary locali- epithelium in vivo. Furthermore, we will monitor the effects
zation of both proteins. To test if SUMOylation is necessary of PI signaling on the electrophysiological output of OSNs
for trafficking of signaling proteins in vivo, mice were dually by patch clamp technique of single neurons in acute OE
infected with wildtype or mutant adenovirus constructs along slices of mOR-EG-GFP transgenic mice.
with Arl13b:mCherry (a cilia marker). Live, en face imag-
ing of OSNs showed wildtype AC3:GFP co-localized with
Ar1l3b:mCherry in olfactory cilia, while the mutant form #P50 POSTER SESSION I: MULTIMODAL
failed to enter olfactory cilia. Surprisingly however, both RECEPTION; CHEMOSENSATION AND DISEASE;
wildtype and mutant ANO2:GFP trafficked into olfactory OLFACTION PERIPHERY
cilia, indicating potential other mechanisms permitting traf-
ficking in vivo. In addition, the generation of SUMOylation Phospholipase C Mediates Intracellular Ca2+ Increase
sites in the related channel, ANO1 was not sufficient for cili- via Internal Ca2+ Stores and trpM5 Activation in Mouse
ary entry. Together our data demonstrate that SUMOylation Olfactory Sensory Neurons
of some signaling proteins is necessary, but not sufficient for Steven A. Szebenyi, Tatsuya Ogura, Jen Chang, Aaron
ciliary localization. Acknowledgements: This work was sup- Sathyanesan, Weihong Lin
ported by NIDCD grants 1R01DC009606-01 (JRM) and
Department of Biological Sciences, University of Maryland,
1F32DC011990-01 (JCM)
Baltimore County, Baltimore, MD, USA
Phospholipase C (PLC) and internal Ca2+ stores are involved
#P49 POSTER SESSION I: MULTIMODAL in a variety of cellular signaling including sensory transduc-
RECEPTION; CHEMOSENSATION AND DISEASE; tion. However, our understanding of the PLC pathway in
OLFACTION PERIPHERY mammalian olfactory sensory neurons (OSNs) is largely
limited to those studies in which the PLC inhibitor U73122
Functional characterization of alternative signal was used to suppress odor responses. Recently, transient
transduction pathways in olfactory receptor neurons receptor potential channel M5 (trpM5) has been shown to
Paul Scholz, Sabrina Baumgart, Katharina Klasen, Benjamin express in a population of mature OSNs in mice (Lin et al
Kalbe, Hanns Hatt 2007). trpM5 is an essential downstream effector of the
PLC pathway for taste transduction. Here we investigate
Ruhr-University-Bochum Bochum, Germany
PLC, trpM5 and internal Ca2+ stores in freshly isolated
It is generally agreed that in olfactory sensory neurons mouse OSNs using single cell Ca2+ imaging. We found that
(OSNs) the binding of odorant molecules to their specific OSNs responded to a PLC activator m-3M3FBS in a con-
olfactory receptor (OR) triggers a cAMP-dependent centration dependent manner with a higher percentage of
signaling cascade activating cyclic-nucleotide gated (CNG) responding cells and greater amplitudes after an increase in
channels. However, considerable controversy dating back concentration (78%, n=23 at 15µM, 90.3%, n=52 at 25µM).
more than 10 years has surrounded the question of whether In contrast, only one out of 9 OSNs responded to the inac-
phosphoinositide (PI) signaling plays a role in mammalian tive analog o-3M3FBS (25µM). Eliminating extracellular
olfactory transduction. Early studies of PI signaling in Ca2+ did not reduce the percent of responding OSNs to
olfaction focused solely on the classical phospholipase C m-3M3FBS and only the response amplitudes were moder-
(PLC) dependent pathway, demonstrating that odorants ately reduced (n=7). In addition, The PLC inhibitor U73122
can elevate inositol triphosphate (IP3). In addition, our (5–10µM) greatly reduced the percent of OSNs responding
recent study proved that odorants stimulate both, PLC and to m-3M3FBS (37.5%, n=8) and the response amplitudes.
phosphatidylinositol 3-kinases (PI3Ks) in the dendritic Further, using OSNs isolated from trpM5-GFP and trpM5
knobs and in olfactory cilia of rodent OSNs. In this project, knockout-GFP mice, we found that trpM5-expressing OSNs
we aim at characterizing the dual pathway of olfactory responded to m-3M3FBS with a significantly larger ampli-
signaling in more detail. For this purpose, we will analyze the tude and calcium load than the trpM5- null OSNs (n= 6
distribution of PI signaling upon specific odor stimulation to 9 for each group). Our data suggest that most OSNs are
in living OSNs via translocation imaging. The use of mOR- capable of utilizing the PLC pathway to release Ca2+ from
EG-GFP transgenic mice will allow for specific analysis of internal Ca2+ stores and that subsequent activation of trpM5
OSNs expressing the well-characterized olfactory receptor in trpM5-expressing OSNs leads to additional increases in
mOR-EG, which can be activated by different odorants. intracellular Ca2+ loads. Acknowledgements: Supported by
research grants NIH/NIDCD 009269, 012831 and ARRA evidence supports a role for these receptors in extra-olfactory
administrative supplement to WL functions. We have examined expression of the odorant
receptors M71, M72, I7, P2, MOR28, whose expression
patterns have been well characterized in the olfactory
#P51 POSTER SESSION I: MULTIMODAL system. We screened for expression of these receptors in
RECEPTION; CHEMOSENSATION AND DISEASE; extra-olfactory tissues by RT-PCR and then tested positive
OLFACTION PERIPHERY results by in situ hybridization, immunohistochemistry,
Analysis of the novel protein Q8BH53 in olfactory sensory or through the use of established reporter mouse lines.
neuron cilia Identification of novel expression outside the olfactory
system suggests a function for odorant receptors in these
Anna K Talaga1, Aaron B Stephan2, Varun Chokshi1, tissues. Future research will be aimed toward uncovering
Haiqing Zhao1 functions and identifying putative endogenous ligands.
1
Johns Hopkins University/Biology Baltimore, MD, USA, Acknowledgements: NIH DC007395
2
UCSD/Division of Biological Sciences La Jolla, CA, USA
The cilia of olfactory sensory neurons (OSNs) are spe- #P53 POSTER SESSION I: MULTIMODAL
cialized for encoding and transducing odor information. RECEPTION; CHEMOSENSATION AND DISEASE;
Among the proteins found in the cilia, many are critical OLFACTION PERIPHERY
for mediating and/or modulating olfactory signal trans- M3-R inhibits β-arrestin2 recruitment and desensitization of
duction. We detected a novel protein, Q8BH53, from a mammalian odorant receptors to potentiate their activation
proteomic screen of OSN cilial membrane preparations.
Yue Jiang1, Yun R. Li2, Hiroaki Matsunami1,3
Q8BH53 is conserved among eukaryotes and is a unique
protein, as no other paralogs exist in the mouse genome. 1
Duke University Durham, NC, USA, 2University of
Bioinformatic analysis suggested that the majority of Pennsylvania, Perelman School of Medicine Philadelphia,
the Q8BH53 sequence is composed of ARM- domains. PA, USA, 3Duke University Medical Center Durham, NC, USA
Although the function of Q8BH53 is unknown, the pres- Adjusting sensitivity of sensory stimuli needed by an ani-
ence of these ARM-repeat domains signifies that it may mal at any given time is crucial for animals’ survival. In
be important for establishing protein-protein interactions. mammals, the activation of odorant receptors (ORs), which
q8bh53 transcripts are abundant in the mouse olfactory epi- are expressed in the olfactory sensory neurons (OSNs) in
thelium and are also found in several other tissues. Using the olfactory epithelium, mediates the perception of smell.
immunohistochemistry, we found that Q8BH53 localizes These ORs belong to the large family of G protein-cou-
specifically to OSN cilia but is largely excluded from the pled receptors (GPCRs), which also include the muscarinic
respiratory epithelium cilia in adult mice. Furthermore, we acetylcholine receptors that are key mediators of the para-
found that Q8BH53 expression begins around embryonic sympathetic nervous system output. Previously, we showed
day E13.5 in the olfactory epithelium. We are currently that activation of the muscarinic receptor M3-R has been
using a knock-out mouse model to understand the func- shown to potentiate OR-mediated cAMP response, and the
tional role of Q8BH53 in the olfactory system. In addition, functional interaction between the M3-R and ORs suggests
we are taking a biochemical approach to identify binding that odorant detection may be modulated by the neuro-
partners of Q8BH53 in OSN cilia. Acknowledgements: transmitter at the peripheral level. However, the mecha-
NIH DC007395 nisms underlying this modulation were not understood.
Here we provide evidence suggesting that M3-R mediates
#P52 POSTER SESSION I: MULTIMODAL the potentiation of OR signaling by inhibiting the recruit-
RECEPTION; CHEMOSENSATION AND DISEASE; ment of β-arrestin-2 to activated ORs. In line with this, acti-
OLFACTION PERIPHERY vation of the M3-R by muscarinic agonist further inhibits
β-arrestin-2 recruitment to ORs, while M3-R antagonist
Expression of Several Odorant Receptors Outside the alleviates the inhibition. These effects are not explained
Olfactory System by the competition for β-arrestin-2 between the two recep-
Crystal M. Wall, Marsalis Brown, Haiqing Zhao tors. Further more, the third intracellular loop of the M3-R
is responsible for its regulation of OR activity. This data
Johns Hopkins University/Biology Baltimore, MD, USA
suggests that the M3-R potentiates OR- mediated cAMP
Over one thousand G-protein coupled receptors have been response largely by inhibiting the β-arrestin-2 recruitment
classified as odorant receptors in the mouse genome based to ORs, providing evidence for a novel mechanism of OR
on sequence similarities with receptors found in the olfactory activity regulation by non-OR GPCRs, with β-arrestin-2 as
system. Recent studies have identified odorant receptors a crucial mediator. Acknowledgements: This work is sup-
expressed outside of the olfactory system, and growing ported by grant from NIH.
#P54 POSTER SESSION I: MULTIMODAL performed have been directed towards discovering whether or
RECEPTION; CHEMOSENSATION AND DISEASE; not the same results are observed in mice that were mutated
OLFACTION PERIPHERY with a lecithin retinyl acyl transferase (LRAT knockout, KO)
gene. To determine the VAD effects on mice, specifically on
Off-flavor substances in foods and beverages cause a ORN numbers we took LRAT KO male mice who were age-
potent suppression of olfactory signal transduction matched and fed them a VAD diet or a diet supplemented
Hiroko Takeuchi1, Hiroyuki Kato2, Takashi Kurahashi1 with VA (VAS) for 8 weeks (Study 1) or 19 weeks (Study 2).
Studies included our control group, age matched, male wild
1
Osaka University Osaka, Japan, 2Daiwa Can Company
type (WT) VAS mice. Our VAD rats exhibit distinct signs of
Tokyo, Japan
VAD that include alopecia, ataxia, reddened eyes, white teeth
We examined the effect of off-flavors in foods/beverages and decreased weight gain relative to controls, LRAT KO
on the olfactory receptor cells under the whole-cell voltage mice did not display any of these signs after eight weeks on
clamp recording configuration. Generally, it has been shown the VAD diet. LRAT KO mice on the VAD diet for 19 weeks
that off-flavor substances induce exogenous unpleasant showed a relative weight loss in the latter part of the study.
smells even with a very low concentration (ppt level inclusion Cytosolic extracts from Olfactory tissue were collected from
in products). Although not yet scientifically demonstrated, mice in Study 2. Immunoblots were prepared and probed
it has also been pointed out that off-flavors reduce the with an antibody directed against olfactory marker protein
pleasant flavors contained in foods/beverages. One of the (OMP), a marker for mature ORNs. Chemiluminescent
most powerful off- flavors is 2,4,6-trichloroanisole (TCA), reagent detection system and images were acquired with a
that is especially known for inducing the corktaint of wines. BioImaging System were used to evaluate relative OMP pro-
In the present study, we show with human psychophysical tein expression levels in LRAT KO VAD, LRAT KO VAS,
tests that TCA and related substances actually reduce and WT mice olfactory tissue cytosolic lysates. Signal area
flavors of foods and beverage with very low concentration. densities were recorded using an EpiChemi Darkroom UVP
It was shown that TCA also suppressed cyclic nucleotide- Bioimaging System. This allowed us to quantify the den-
gated (CNG) channels potently, when examined in olfactory sity of each band on the immunoblot. Results have shown
sensory cilia. Surprisingly, the channel suppression was that VAD mice have less OMP expression than those ani-
detected even when 1 aM of TCA was applied to the cell mals fed a VAS diet including the LRAT KO VAS mice and
with an U-tube system. To explain such super- efficiency, the the WT mice. These findings suggest that VAD may have
TCA effect showed the time-integration and slow recovery different and/or less pronounced effects on mice than rats.
from the current suppression, presumably representing the Acknowledgements: NIH/NIGMS/MBRS/ SCORE S06
integration of the substance into the hydrophobic site of the GM 008092
membrane. Based on the relation between the number of
TCA molecules applied and total number of CNG channels,
it was assumed that single TCA molecule may affect more #P56 POSTER SESSION II: OLFACTION
than one CNG channel. This is also consistent with an idea DEVELOPMENT; TASTE CNS; NEUROIMAGING;
that the effect of TCA is mediated by the lipid bilayer to OLFACTION CNS
affect surrounding channels simultaneously. TCA was found
in a wide variety of foods/beverages, when screened out from Cerebral processing of odors related to their hedonic
their flavor losses. Natural generation of TCA and related judgment
off-flavor substances may be one of the mechanisms for the Anna Blumrich, Cornelia Hummel, Emilia Iannilli, Thomas
deterioration of those products. Hummel Smell & Taste Clinic
Department of Otorhinolaryngology, University of Dresden
Medical School Dresden, Germany
OLFACTION PERIPHERY The aim of the study was to investigate parameters reflecting
the hedonic component of odor perception. The emotional
Effect of Vitamin A Deficiency on Olfactory Marker Protein effect of smelling- claiming an odor as pleasant or unpleasant-
Expression in Postnatal Mouse Olfactory Neurons is an important part of the central nervous connection
Bukola A. Oke, Mary Ann Asson-Batres of odor perception. It has been shown that perception of
pleasant odors implicates different cerebral activations than
Tenneessee State University Biology Department Nashville,
that of unpleasant ones. Thirty-two healthy, right-handed
TN, USA
subjects (16 men, 16 women; mean age 23.4 years; range
Our lab has previously shown that Olfactory receptor neu- 20–29 years) were examined. Normal olfactory function was
ron (ORN) levels decrease when sources of Vitamin A have ascertained using the “Sniffin Sticks”. All subjects rated peach
been removed from the diet of postnatal rats. Experiments odor as pleasant, and the smell of butanol as unpleasant.
According to their hedonic judgement of liquorice odor, analyses of the behavioral data demonstrate a replication
subjects were divided into two groups: group 1 subjects of the aforementioned findings. As predicted, valence-inde-
described the odor of liquorice as pleasant, and group 2 pendent olfactory-visual integration was mediated by low-
subjects experienced liquorice as unpleasant. Functional level multisensory integration areas in the superior parietal
magnetic resonance imaging (fMRI) was used to compare lobule. Preliminary analyses of the fMRI data indicate that
the cerebral activations while smelling the three different valence-dependent integration occurs in higher-order multi-
odors: pleasant peach, unpleasant butanol, and ambiguous sensory integration areas in conjunction with areas known
liquorice. Analysis indicated common neural activations in to code for odor valence. Moreover, a differential processing
response to all odors in a number of regions, e.g. in cingulate of unpleasant compared to pleasant olfactory-visual stimu-
gyrus, medial frontal gyrus and caudate nucleus. In subjects lus combinations has been established. Further insights into
disliking liquorice, activations were found in four areas the neural processes mediating the influence of valence on
including medial frontal gyrus, postcentral gyrus and medial the neural correlates of olfactory-visual integration will be
temporal gyrus. In liquorice- liking subjects, activations were discussed. Acknowledgements: Funded by a startup grant
found in the medial frontal lobe. These results suggest that from the Medical Faculty of RWTH Aachen University.
there are different patterns of central nervous activation
dependent on the hedonic value of an odor, mainly including
the frontal lobe, with unpleasant odors activating the more #P58 POSTER SESSION II: OLFACTION
lateral frontal lobe and pleasant odors more medial areas. DEVELOPMENT; TASTE CNS; NEUROIMAGING;
In addition, unpleasant odors produced more activation as OLFACTION CNS
compared to pleasant odors. Acknowledgements: Supported
by Takasago, Paris. Visuo-olfactory integration facilitates peri-threshold
olfactory categorization
Jaryd Hiser1, Lucas R. Novak1, Wen Li1,2
#P57 POSTER SESSION II: OLFACTION Department of Psychology, University of Wisconsin -
1
DEVELOPMENT; TASTE CNS; NEUROIMAGING; Madison Madison, WI, USA, 2Waisman Center, University of
OLFACTION CNS Wisconsin - Madison Madison, WI, USA
Valence Modulation of Crossmodal Olfactory-Visual Neural Olfactory quality discrimination and categorization
Integration proves a highly challenging process in humans. Additional
Jessica Freiherr1, Anna-Nora zur Nieden1, Johan information from other senses, such as visual cues, may
N. Lundström2,3,4 facilitate this operation via crossmodal integration. To date,
crossmodal sensory integration has focused on non-chemical
1
RWTH Aachen University, Diagnostic and Interventional
senses. Using functional magnetic resonance imaging
Neuroradiology Aachen, Germany, 2Monell Chemical Senses
(fMRI) techniques, this study characterized visuo-olfactory
Center Philadelphia, PA, USA, 3University of Pennsylvania,
integration in olfactory categorization. Participants (N=29)
Department of Psychology Philadelphia, PA, USA,
smelled an odor at a merely detectable level from one of three
4
Karolinska Institute, Department of Clinical Neuroscience
categories (food, floral, or wood) while viewing a picture that
Stockholm, Sweden
was congruent or incongruent to the odor, and then made a
We recently demonstrated that a concurring congruent vis- category decision on the odor. Reaction time (RT) was faster
ual stimulus does not affect olfactory sensitivity but does for congruent versus incongruent stimuli (P<.05). Similarly,
modulate the perceived valence or pleasantness and inten- accuracy was higher for congruent than incongruent stimuli
sity of an odor. However, the congruency-dependent effect (P’s<.005). These results reveal effective visuo-olfactory
occurred only for odors perceived as pleasant. With the cur- integration in improving otherwise chance-level odor quality
rent study we explored the neural mechanisms of this behav- categorization. fMRI analysis demonstrated enhanced
ioral phenomenon with the aim of determining the influence right olfactory orbitofrontal cortex (oOFC) response to
of valence on the neural correlates of crossmodal olfactory- congruent versus incongruent visuo-olfactory pairing, for
visual integration. To this end, the pleasant odor phenyl ethyl all three odor categories. Importantly, this enhancement
alcohol and unpleasant odor isovaleric acid was applied using in oOFC was strongly associated with congruency-related
constant-flow olfactometry in combination with a congru- improvement in odor categorization (in RT and accuracy).
ent, incongruent, or blank visual stimulus during an event- Our findings suggest that despite the peri-threshold level
related fMRI paradigm. As control stimuli we also applied of odor presentation and consequent difficulty in odor
pleasant and unpleasant visual stimuli and a baseline stimu- categorization, visual input can be integrated with olfactory
lus. We investigated brain activation due to crossmodal inte- information and markedly improve this process. In particular,
gration in 14 healthy, normosmic participants. Subjects had the secondary olfactory cortex in the OFC mediates this
to rate pleasantness of the odors after each event. Statistical process, presumably via its highly associative multimodal
connections. Multi-voxel pattern analysis is underway #P60 POSTER SESSION II: OLFACTION
to examine nuanced quality representation by ensemble DEVELOPMENT; TASTE CNS; NEUROIMAGING;
neuronal activity in the primary (piriform) and secondary OLFACTION CNS
(oOFC) olfactory cortices. Acknowledgements: Funding
supported by R01MH093413 (W.L.), P30HD03352 (W.L.) Smell what you seek – Reward sensitivity amplifies visuo-
olfactory integration of positive affect
#P59 POSTER SESSION II: OLFACTION Lucas R. Novak1, Jaryd Hiser1, Wen Li1,2
DEVELOPMENT; TASTE CNS; NEUROIMAGING; Department of Psychology, University of Wisconsin -
1
OLFACTION CNS Madison Madison, WI, USA, 2Waisman Center, University of

The chemosensory path of pain Wisconsin - Madison Madison, WI, USA
Kathrin Kollndorfer1, Ksenia Kowalczyk1, Johannes Crossmodal integration is a process ubiquitous in animals with
Frasnelli2, Elisabeth Hoche1, Ewald Unger3, Christian multiple sensory systems, facilitating perception especially
A. Mueller4, Siegfried Trattnig5, Veronika Schöpf1 when challenged with limited stimulus input. However, there
is little research on integration of positive affect, still less
1
Medical University of Vienna, Department of Radiology, how this process varies with relevant individual differences
Division of Neuro- and Musculoskeletal Radiology Vienna, such as reward sensitivity. Reward sensitivity may increase
Austria, 2CERNEC, Université de Montréal, Département response to rewarding stimuli, thereby facilitating crossmodal
de Psychologie Montreal, QC, Canada, 3Medical University integration of positive affect. Applying functional magnetic
of Vienna, Center for Medical Physics and Biomedical resonance imaging (fMRI) techniques, this study examined
Engineering Vienna, Austria, 4Medical University of Vienna, visual and olfactory integration of reward. Participants
Department of Otorhinolaryngology Vienna, Austria, (N=29) performed an odor categorization task: they smelled
5
Medical University of Vienna, Department of Radiology, an odor from one of two categories (floral or wood) while
MR Centre of Ecxellence Vienna, Austria viewing an image congruent or incongruent with the odor,
The function of the olfactory system has been well inves- followed by a category decision. Reaction time was faster
tigated over the last decades. However, we know much less and accuracy greater for floral than wood odors (P’s<.005),
regarding central processing of intranasal trigeminal stimuli. and accuracy was greater for congruent than incongruent
Trigeminal sensations like burning, stinging, tingling, pun- trials (P<.005), suggesting preferential response to pleasant
gency and temperature constitute an additional quality in and bimodal stimulation. fMRI analysis demonstrated
perception of odor and flavor, but also allow odor locali- enhanced left amygdala and hippocampus activation for
zation in contrast to pure olfactory odors. In this study we floral than wood odors and congruent than incongruent
focused on three different trigeminal compounds which tar- trials. Although behavioral response was not associated with
get various sensory receptors and produce distinctive sen- reward sensitivity as measured by the Behavioral Activation
sations: CO2 elicits stinging sensations (activating TRPV1 Scale (BAS), positive affective and integrative responses in
receptors), cinnamaldehyde burning sensations (TRPA1) and right amygdala, hippocampus, and piriform cortex were
menthol cooling sensations (TRPM8). The point of interest greater with higher BAS scores, especially in fun-seeking.
was to investigate discrepancies in the processing of different The amygdala and hippocampus are key contributors
intranasal chemosensory trigeminal stimulations along the in the reward circuitry in the brain, projecting to ventral
trigeminal pathway. Moreover, we aimed to reveal activation striatum and triggering downstream processes driving
in olfactory areas which supports a close relationship between reward experience. Higher reward sensitivity thus promotes
the two chemosensory systems. We conducted an fMRI study reward processing by facilitating integrative processing of
on a 3T MRI scanner to measure neural activity in 12 healthy pleasant sensory inputs in both limbic and primary sensory
subjects. Each of the three stimuli was delivered separately by regions. Acknowledgements: R01MH093413 (W.L.) and
a computer-controlled air-dilution olfactometer in one ses- P30HD03352 (W.L.)
sion. Using an event-related design subjects received trigemi-
nal stimuli of 500ms to the left nostril (ISI 30s). During the
whole experiment subjects were asked to breathe using velo- #P61 POSTER SESSION II: OLFACTION
pharyngeal closure. Functional imaging data were analyzed DEVELOPMENT; TASTE CNS; NEUROIMAGING;
with Independent Component Analysis. Insula, SI and OFC OLFACTION CNS
were activated by all three compounds. Further, all three con-
Biochemical components of trigeminal integration in
ditions led to activation of the PFC demonstrating the well
anosmics: A pilot functional magnetic spectroscopy study
established interaction between the olfactory and trigeminal
system. The findings support the notion that activation of the Veronika Schöpf1, Kathrin Kollndorfer1, Elisabeth Hoche1,
three trigeminal receptors is processed in the same pathway. Bernhard Strasser2, Ksenia Kowalczyk1, Ewald Unger3,
Acknowledgements: FWF (P23205-B09) Christian A. Müller4, Siegfried Trattnig2, Martin Krssak2,5
1
Department of Radiology, Division of Neuro- and 4
Department of Psychology, University of Pennsylvania
Musculoskeletal Radiology Medical University of Vienna Philadelphia, PA, USA, 5Department of Clinical Neurosience,
Vienna, Austria, 2Department of Radiology, MR Centre of Karolinska Institute Stockholm, Sweden
Excellence, Medical University of Vienna Vienna, Austria,
The combination of taste and odor found in a flavorful
3
Center for Medical Physics and Biomedical Engineering,
dish creates a more powerful sensation than its odor or
Medical University of Vienna Vienna, Austria, 4Department
taste in isolation. Whereas the neural processing of the
of Otorhinolaryngology, Medical University of Vienna
individual chemosensory components is well known, the
Vienna, Austria, 5Department of Internal Medicine III,
functional connectivity underlying the combined flavor
Division of Endocrinology and Metabolism, Medial
percept is poorly understood. In the present functional
University of Vienna Vienna, Austria
magnetic resonance imaging study, subjects were presented
Several studies investigated functional interaction of the with taste only (gustatory presentation of juice, closed soft
olfactory and trigeminal system. Although anosmic patients palate), smell only (orthonasal presentation of juice odor),
are not able to perceive odors they show changes in “insu- or a combined flavor (retronasal-gustatory presentation,
lar” blood oxygenation during trigeminal neuronal activity. swallowing juice). As expected, olfactory stimulation alone
This pilot study aimed to investigate how changes on blood activated olfactory areas while gustatory stimulation alone
oxygenation level, as observed by fMRI, are underlined by elicited activation within the gustatory cortex. Overlapping
changes in neurotransmitter (GABA, glutamate) balance activation within both networks could be observed during
under trigeminal simulation in the insula and how these flavor presentation, and a convergence zone between all
changes differ between anosmic and healthy populations. 3 three conditions was observed in the anterior ventral insula
(2f;25–43ys) functional anosmics and 8 controls (6f;18–43ys) and cingulate cortex. Superadditive activity for the flavor
were examined using proton magnetic resonance spectros- condition, relative to odor and taste alone, was observed in
copy (1H-MRS) to explore changes of excitatory and inhibi- the dorsal insular gyrus, extending into parietal operculum
tory neurotransmitters in the insula. Spin-echo based MRS and postcentral gyrus. Finally, to delineate the cerebral
(TE=30ms/TR=5000ms) measurements were performed on a networks contributing to the flavor percept, we assessed
3T whole body MR scanner, using a 32ch coil for signal detec- the functional connectivity between these significant nodes
tion combined with a stimulation device, which was designed responsive to chemosensory overlap during combined odor-
specifically for intranasal application. The paradigm consisted taste stimulation. Increases in functional connectivity with
of 4 stimulation blocks: 4 dynamic cycles with 32 acquisitions, both convergent and superadditive areas were observed in
and 32 stimuli (CO2, 50%v/v, birhinal, 250ms). Acquired an overlapping area in the temporal pole. Taken together,
spectral transients were individually frequency corrected, these findings are suggestive of an important relay function
phased and further grouped according to the timeline posi- of semantic memory circuits in the formation of the flavor
tion into baseline- and stimulation-groups. The quantification experience from crossmodal chemosensory information.
of metabolic intensities was performed using the LCModel
with an imported modelled basis set including metabolites #P63 POSTER SESSION II: OLFACTION
and macromolecular resonances. Results for controls revealed
DEVELOPMENT; TASTE CNS; NEUROIMAGING;
decreased GABA (41.24%) during the rest phase. Anosmic
OLFACTION CNS
patients showed a significant decrease of glutamate (16.42%)
and a higher GABA response (205.25%) rate to stimulation Is the Superior Temporal Sulcus involved in the Reinforced
compared to controls. Results of this study will significantly Configural Processing of a Binary Odor Mixture?
contribute to the basic understanding of trigeminal process- Charlotte Sinding1,2, Gérard Coureaud1, Noëlle Béno1, Elodie
ing of chemosensory information in patients with olfactory Le Berre1, Cornelia Hummel2, John Prescott3, Moustafa
dysfunction. Acknowledgements: FWF (P23205-B09) Bensafi4, Thomas Hummel2, Thierry Thomas-Danguin1
1
Centre des Sciences du Goût et de l’Alimentation (CSGA),
#P62 POSTER SESSION II: OLFACTION UMR 6265 CNRS, UMR 1324 INRA, Université de Bourgogne,
DEVELOPMENT; TASTE CNS; NEUROIMAGING; Developmental Ethology and Cognitive Psychology Team
OLFACTION CNS and Flavour Perception Team Dijon, France, 2Smell & Taste
Clinic, Department of Otorhinolaryngology, University of
Superadditive processing during flavor perception is
Dresden Medical School Dresden, Germany, 3TasteMatters
modulated by anterior temporal cortex connectivity
Research & Consulting Sydney, Australia, 4Centre de
Janina Seubert1, Kathrin Ohla1,2, Yoshiko Yokomukai3, Johan Recherche en Neurosciences de Lyon, UMR5020 CNRS,
N. Lundström1,4,5 Université Lyon 1, Neurosciences Sensorielles Comportement
Monell Chemical Senses Center Philadelphia, PA, USA,
1 Cognition Lyon, France
German Institute of Human Nutrition Potsdam-Rehbrücke,
2
In macaque monkeys the superior temporal sulcus (STS)
Germany, 3Kirin Holdings Company LTD Tokyo, Japan, projects to the orbitofrontal cortex which projects to the
primary olfactory cortex (Carmichael and Price 1995). associated weight gains. An understanding of the neural
These connections are believed to be reciprocal, therefore processes of taste perception is essential for elucidating this
the STS is supposed to receive inputs from olfactory areas. disease state. Unfortunately, studies on the human brain
Kettenmann et al. (1996) showed that STS was activated are limited, and thus the majority of neural taste function
when participants were stimulated with monomolecular knowledge stems from animal studies. These studies have
odors. This area is also specifically activated for configural shown that hedonic and intensity information is encoded in
visual processing, as during the perception of body motion the primary gustatory cortex, with neuronal firing correlated
(Thompson et al. 2005). In the present study, we investigated with stimulation intensity. Seeking to confirm animal models
the configural processing of odor mixtures in human adults. in the human brain, this study utilized fMRI to study BOLD
We repeatedly exposed (2 sessions of 11 exposures) healthy signal changes in response to varied concentrations of sweet
volunteers (n=12, GAB) to a binary mixture (AB) configu- and salty taste stimulants. Normal, healthy volunteers
rally processed (blending of the two components’ odors into (n=11) completed a total of fifteen taste fMRI studies in
a single pineapple odor), while others (n=14, Gcompo) were which brain response to an event-related taste stimulation
exposed to the separate components, A (“strawberry”) and paradigm was correlated with perceived ratings of both
B (“caramel”). To equilibrate the number of exposures in pleasantness and intensity. The data indicate a positive
the two groups, GAB was also exposed to PEA (monomo- correlation between pleasantness and intensity for sweet
lecular, “rose”). Such exposures were known to favor the tastants; while a negative correlation was observed for salty
perception of the AB configuration in GAB and the percep- tastants. Each triggered significant activation in the primary
tion of the elements in Gcompo (Sinding et al. 2011, in and secondary gustatory cortices including: bilateral anterior
preparation). Two days after the pre-exposure, all subjects insular cortex, posterior orbitofrontal cortex, cingulated
received an fMRI while stimulated by AB, A, B and PEA. cortex, and dorsolateral prefrontal cortex. BOLD signals in
As a major result, the STS appeared significantly more acti- these regions were significantly correlated with both hedonic
vated for the processing of the AB mixture in GAB than in and intensity ratings from subjects. Overall these results
Gcompo. The STS was also more activated for the process- show that the human brain processes hedonic and intensity
ing of AB as compared to A and B, in GAB. The STS was information of sweet and salty taste through similar neural
not significantly activated in Gcompo for any stimulation, in networks previously seen in animal models
any contrast. These results suggest that the STS is a criti-
cal area for the reinforced configural processing of simple
odorant mixtures. However, PEA also activated significantly #P65 POSTER SESSION II: OLFACTION
the STS in GAB in comparison to Gcompo. Acknowledgements: DEVELOPMENT; TASTE CNS; NEUROIMAGING;
Supported by grants from the Burgundy Regional council OLFACTION CNS
and EU-ERDF to GC and TTD, European Dijon-Dresden
Laboratory (LEA 549) to TH, GC, TTD, and a fellowship Perception and encoding of odor elements and mixtures in
from the French MESR to CS. the human brain
Keng Nei Wu1, Sydni M. Cole1, Jay A. Gottfried1,2
1
Northwestern University/Neurology Department, Feinberg
#P64 POSTER SESSION II: OLFACTION School of Medicine Chicago, IL, USA, 2Northwestern
DEVELOPMENT; TASTE CNS; NEUROIMAGING; University/Department of Psychology Evanston, IL, USA
OLFACTION CNS
In the natural environment, most odorous objects are com-
Association of Pleasantness and Intensity of Sweet and posed of dozens, if not hundreds, of volatile molecules.
Salty Taste in the Human Brain Despite this apparent complexity, the olfactory system seam-
Jianli Wang1, Sebastian Rupprecht1, Zachary Mosher1, Robert lessly integrates these components into perceptual wholes.
Mchugh1, Jeffrey Vesek1, Sarah Ryan1, Megha Vasavada1, Utilizing a between subject design, this experiment aimed
Qing X. Yang1,2, Andras Hajnal3,4, Ann M. Rogers4 to investigate how experience in the form of aversive learn-
ing modulates perception and encoding of odor mixtures by
1
Penn State College of Medicine/Radiology Hershey,
pairing either a target binary odor mixture (Mx) or one of
PA, USA, 2Penn State College of Medicine/Neurosurgery
its components (Ele) with an electric shock. We used psy-
Hershey, PA, USA, 3Penn State College of Medicine/Neural &
chophysical measurements, functional magnetic resonance
Behavioral Sciences Hershey, PA, USA, 4Penn State College of
imaging (fMRI), and multivariate analytical techniques to
Medicine/Surgery Hershey, PA, USA
investigate these learning induced changes. We presented
An abnormal assessment of pleasantness and intensity of human subjects with six stimuli: three monomolecular odor-
tastants can lead to negative, long-term health conditions. ants (A, B, C), and three binary mixtures (AB, BC, AC).
Among these, obesity may be a consequence of diminished To date, results have been collected for 13 subjects (8 Ele,
sensitivity to sweet tastes, leading to amplified cravings and 5 Mx) who were successfully conditioned. When asked to
identify the odor component(s) of these stimuli, subjects group but not in the anosmic group. This hedonic specific
in the Mx group showed decreased accuracy in identifying approach was meant to control for activation that was due to
the correct component(s). Moreover, these subjects also attention allocation or activation of semantic circuits that are
rated mixtures to be less similar to their components, while alone sufficient to evoke activation in olfactory areas. Direct
subjects in the Ele group rated mixtures to be more similar comparisons between the groups revealed greater activation
to their components after conditioning. These preliminary in the anosmic group in olfactory areas than in the control
findings suggest that olfactory learning of a binary mixture group. We conclude that, in contrast to the control group,
may induce perceptual and neural fusion of odor elements anosmic participants have difficulties to perform olfactory
into a synthetic whole. Conversely, pairing a shock with a imagery in the conventional meaning.
component of a binary mixture may induce neural “fission”
of the mixture, such that its components are processed in
a more elemental fashion. Ongoing fMRI analysis will test #P67 POSTER SESSION II: OLFACTION
the hypothesis that learning induced changes in odor qual- DEVELOPMENT; TASTE CNS; NEUROIMAGING;
ity perception may be reflected in the correlation between OLFACTION CNS
odor evoked patterns of activation in the posterior piriform
cortex. Acknowledgements: This work was supported by Multi-modal functional imaging of rat olfactory bulb with
Northwestern Institutional Predoctoral Training Awards orthonasal and retronasal odorant stimulation: functional
to K.N.W. (T32NS047987) and grants R01DC010014 and insights through complementary techniques
K08DC007653 from the US National Institute on Deafness Michelle R Rebello1,2, Basavaraju G Sanganahalli3, Gordon M
and Other Communication Disorders to J.A.G. Shepherd1,2, Fahmeed Hyder3, Justus V Verhagen1,2
1
The John B. Pierce Laboratory New Haven, CT, USA, 2Yale
School of Medicine, Dept. Neurobiology New Haven, CT,
#P66 POSTER SESSION II: OLFACTION USA, 3Yale University, MRRC New Haven, CT, USA
OLFACTION CNS Various techniques can be used to evaluate odor response
maps of the olfactory bulb, each with its own advantages.
The Fate of the Inner Nose: Odor Imagery in Patients With Here we combined fMRI with intrinsic and calcium opti-
Olfactory Loss cal imaging to better characterize glomerular responses.
Elena L. R. Flohr1,2, Artin Arshamian3,1, Matthias J. Wieser2, Functional MRI and intrinsic imaging share slow responses
Cornelia Hummel1, Maria Larsson3, Andreas Muehlberger2, of complex origin which are based on dynamic changes in
Thomas Hummel1 oxy- and deoxy-hemoglobin. Whereas fMRI can image the
entire bulb, intrinsic and calcium are limited to the dorsal
1
Smell and Taste Clinic, University of Dresden Medical School
regions but with high spatiotemporal resolution. CBV-
Dresden, Germany, 2Department of Psychology I, University
weighted fMRI was unsuccessful in bulb imaging, as was
of Würzburg Wuerzburg, Germany, 3Department of
the use of other anesthetics (Ex:domitor). In seven rats we
Psychology, Stockholm University Stockholm, Sweden
performed micro fMRI (BOLD, 120x120x300µm) in dorsal
Although the concept of olfactory mental imagery remains orientation so that data could be compared to subsequent
controversial, recent studies support the principle. Cerebral calcium imaging of orthonasal responses in the same sub-
activations during olfactory mental imagery are fairly well jects. In a few cases odor induced activation maps showed
investigated in healthy participants but very few studies strong overlap between the two imaging modalities. We fur-
address the subjects of olfactory imagery in patients with ther apply these techniques in separate animals to define
olfactory loss. To investigate if olfactory imagery is impaired retronasal dorsal and whole bulb responses and compare
in patients who are no longer able to smell, 16 participants those to orthonasal odorant presentations. We have found
with acquired anosmia and 19 normosmic control participants significant effects of odor route on the response magni-
have been investigated. We used functional magnet resonance tude and timing. Further, retronasal odor concentration
tomography and subjective ratings to explore the mechanisms affects response latency in a direction depending on the
during mental imagery of odors. After an imagery training, odorant. We are evaluating several methods of post-hoc
participants imagined odors triggered by words naming co-registering across these methods 1) across functional
pleasant and unpleasant olfactory objects. We found that the maps (fMRI, intrinsic, calcium), 2) using phantom markers
patients compared to healthy control participants showed on the skull, 3) using vasculature via venogram MRI, and
greater difficulties in imagining odors and lower intensity 4) using SWIFT-MRI for skull imaging. Due to the partial
scores while doing so. Looking at neural activation, the volume effect and the relatively thin dorsal glomerular layer
pattern observed by Bensafi et al. (2007) that imagining we are also developing a miniature phased coil-array allow-
unpleasant odors leads to more activation in olfaction-related ing higher resolution and high quality dorsal functional
areas than imagining pleasant odors was found in the control images. Our comparative approach shows the challenges in
obtaining and interpreting odor maps using different meth- 1

Maria-von-Linden-Schule, Heckentalstraße 86 89518
odologies. Acknowledgements: This work is supported by Heidenheim, Germany, 2Faculty of Medicine, Institute of
NIH/NIDCD Grants R01DC009994 and R01DC011286. Anatomy, Department of Neuroimmunology, University of
Leipzig, Liebigstr. 13 04103 Leipzig, Germany
#P68 POSTER SESSION II: OLFACTION The olfactory bulb is the anterior part of the brain and
phylogenetically one of the oldest brain structures. During
postnatal development, when the animal grows, the brain
OLFACTION CNS
increases in size – and so does the olfactory bulb. However,
Quantifying Bursting Olfactory Neuron Activity from in some mammals, such as the American mink (Neovison
Calcium Signals Using Maximum Entropy Deconvolution vison) it is known, that brain shows an overshoot develop-
In Jun Park1, Yuriy V. Bobkov2, Barry W. Ache2,3, Jose ment postnatally with a subsequent reduction in size. Thus
C. Principe1 we were interested, if this applies also to the olfactory bulb.
Therefore we analyzed morphometrically a total of 57
1
Dept. of Electrical and Computer Engineering, University of female minks ranging from newborn (postnatal day 0, P0)
Florida Gainesville, FL, USA, 2Whitney Laboratory, Center for to one year of age for their brain and olfactory bulb size.
Smell and Taste, and McKnight Brain Institute St Augustine, The results reveal, that the volume of one olfactory bulb
FL, USA, 3Depts. of Biology and Neuroscience, University of in newborns is 1.26 ± 0.02 mm3, increasing continuously
Florida Gainesville, FL, USA (P30: 40.27 ± 8.77 mm3; P90: 84.70 ± 1.87 mm3) to adult val-
Advances in calcium imaging have enabled studies of the ues (107.19 ± 4.15 mm3) with no overshoot phenomena. In
activity dynamics of both individual neurons and neuronal contrast, the brain weight increases postnatally from P0
assemblies. However, inferring action potentials (spikes) (0.29 ± 0.06 g) up to P90 (10.18 ± 0.42 g) when maximal val-
from calcium signals is still a challenging issue due to hidden ues are reached, and decreasing afterwards more than 17%
nonlinearity in their relationship, contamination by noise, and to the adult size (8.43 ± 0.35 g). The olfactory bulb growth
often the relatively low temporal resolution of the calcium therefore does not parallel the total brain growth but shows
signal compared to the time-scale of spike generation. Complex an allometric growth pattern. On the other hand, the overall
neuronal discharge, as in the case of the bursting or rhythmically body growth increases continuously to adult values result-
active neuronal activity represents an even greater challenge for ing in an olfactory bulb/body weight ratio of similar val-
reconstructing spike trains based on calcium signals. Here we ues among newborns, brain overshoot age and adults (P0:
propose doing this using blind calcium signal deconvolution 0.014 ± 0.002%; P90: 0.012 ± 0.002%, adult: 0.011 ± 0.001%)
based on a theoretical information approach. The basic idea is with higher values early postnatally (P30: 0.047 ± 0.013%).
to maximize the output entropy of a nonlinear filter where the This indicates that the olfactory bulb is influenced by other
nonlinearity is defined by the cumulative distribution function factors than the cortical neurons for its neuronal network
of the spike signal. We tested this maximum entropy (ME) growth and controlled by different stimuli for its formation
algorithm on bursting olfactory receptor neurons (bORNs) in and connectivity. Further, no postnatal reduction in size
the lobster olfactory organ. The advantage of the ME algorithm suggests a basic and important functional relevance of the
is that the filter can be trained online based only on the statistics olfactory bulb.
of the spike signal without making any assumptions about the
spike-calcium signal relation. We show that the ME method is
able to reconstruct the timing of the first and the last spike of
#P70 POSTER SESSION II: OLFACTION
a burst with higher accuracy compared to other methods. Thus DEVELOPMENT; TASTE CNS; NEUROIMAGING;
the ME method should be a useful tool for inferring parameters OLFACTION CNS
of bursting neurons, including bursting olfactory neurons, Effect of kamikihito (TJ-137) on nerve growth factor and
to help further understand the mechanism and function of olfactory nerve in vivo
bursting-based neuronal sensory coding. Acknowledgements:
Junpei Yamamoto1, HIdeaki Shiga1, Kohshin Washiyama2,
Supported by award R21 DC011859 from the NIDCD
Ryohei Amano2, Takaki MIwa1
1
Otorhinolaryngology, Kanazawa Medical University
#P69 POSTER SESSION II: OLFACTION Ishikawa, Japan, 2Quantum medical technology, Kanazawa
University Ishikawa, Japan
OLFACTION CNS Background: A Kampo product, kamikihito (product name
code: TJ-137), has ingredients that promote nerve growth.
Allometric Growth of Olfactory Bulb and Brain in
Paclitaxel, a cancer chemotherapeutic agent, is toxic to
Female Minks
olfactory nerve cells in vivo. To show TJ-137 pre-medication
Willi Bennegger1, Elke Weiler1,2 effect on nerve growth factor (NGF) in olfactory bulb and
paclitaxel induced olfactory neuropathy in vivo. Methods: VNO neurogenic border. These data suggest that the role
Female 7-week-old Bulb/c mice were fed food containing played by FGF8 in controlling cell fate specification and
TJ-137, or control food, for 14 days before and after neural patterning of the olfactory pit is in large part indirect
intravenous paclitaxel administration. NGF was assessed as Fgf8 levels affect both BMP and Noggin expression in
with ELISA in the olfactory bulb of mice fed food containing the pit and nasal mesenchyme. BMP and Noggin expression
TJ-137 (n=9), or control food (n=9) for 14 days. The epithelial respectively define the epithelial or neurogenic permissive
changes in the nasal turbinates of mice were assessed by borders. The previously described lack of GnRH neuron
H&E and immunohistochemical staining for the olfactory specification in animals with chronically reduced FGF8
marker protein (OMP). The accumulation of the neuronal reflects the loss of a large neurogenic permissive milieu that
tracer (Dextran tetramethylrhodamine) in the olfactory includes the entire VNO. Acknowledgements: The work pre-
bulb was assessed in frozen sections of mice 48 h after nasal sented was supported by the Intramural Research Program
administration of the tracer. Results: NGF was significantly of the National Institutes of Health, National Institute of
increased in the olfactory bulb of mice fed food-containing Neurological Disorders and Stroke
TJ-137 than in the control mice (P=0.017). The epithelium of
nasal turbinates of TJ-137 treated mice was less injured than
that of the control mice after paclitaxel administration. The #P72 POSTER SESSION II: OLFACTION
accumulation of the neuronal tracer in the olfactory bulb DEVELOPMENT; TASTE CNS; NEUROIMAGING;
was higher in the TJ-137 treated mice compared to controls. OLFACTION CNS
Conclusion: We found that TJ-137 is effective in increasing
NGF in olfactory bulb and reducing paclitaxel induced An IP3R3- and NPY-expressing microvillous cell mediates
olfactory neuropathy in vivo. Acknowledgements: Assist tissue homeostasis and regeneration
Kaken from Kanazawa Medical University (2012) Colleen C. Hegg1,2,3, Cuihong Jia1, Sebastien Hayoz1, Chelsea
R. Hutch2,3, Apryl E. Pooley2, Tania R. Iqbal2
1
Michigan State University Pharmacology and Toxicology
#P71 POSTER SESSION II:OLFACTION East Lansing, MI, USA, 2Neuroscience Program East Lansing,
DEVELOPMENT; TASTE CNS; NEUROIMAGING; MI, USA, 3Center for Integrative Toxicology East Lansing, MI,
OLFACTION CNS USA
Fgf8 Defines Neurogenic Vomeronasal and GnRH Calcium-dependent release of neurotrophic factors plays
Neurogenic Mileu by Influencing BMP and BMP Antagonists an important role in the maintenance of neurons, yet
Expression. the release mechanisms are understudied. The inositol
Paolo E. Forni1, Kapil Bharti2, Susan Wray1 triphosphate (IP3) receptor is a calcium release channel that
has a physiological role in development, sensory perception,
1
National Institute of Neurological Disorders and Stroke/NIH
neuronal signaling and secretion. In the olfactory system,
Bethesda, MD, USA, 2National Eye Institute/NIH Bethesda,
the IP3 receptor subtype 3 (IP3R3) is expressed exclusively
MD, USA
in a microvillous cell subtype that is the predominant
FGF8 plays a pivotal role in development of craniofacial cell that expresses neurotrophic factor neuropeptide Y
structures, the olfactory/vomeronasal system and GnRH (NPY). We hypothesized that the IP3R3− expressing
neurons. BMPs can antagonize FGFs expression and sig- microvillous cells secrete sufficient NPY needed for both
nal, BMP and FGF8 signaling are known to exert opposite the continual maintenance of the neuronal population and
roles in defining epithelial versus neurogenic fate. We ana- for neuroregeneration following injury. We addressed this
lyzed the relation between Fgf8, BMP and BMP anatgonists question by assessing the release of neurotrophic factor NPY
in the developing olfactory pit in two Fgf8 hypomorph and regenerative capabilities in wild type mice, IP3R3+/−, and
mouse models, expressing different levels of FGF8. In both IP3R3−/− mice. Injury, simulated using extracellular ATP
mutant mouse models, Fgf8neo/neoand Fgf8neo/Null, regardless as a model, induced IP3 receptor-mediated NPY release
of the FGF8 dosage, overlapping defects were observed in in wild-type mice. ATP-evoked NPY release was impaired
the olfactory pit: lack of neurons formation limited to the in IP3R3−/− mice, suggesting that IP3R3 contributes to
ventral area of the developing nasal pit and the proximal NPY release following injury. Under normal physiological
portion of respiratory epithelium, where GnRH neurons conditions, both IP3R3−/− mice and explants from these mice
normally form. Analyzing expression of BMP4 and BMP had fewer progenitor cells that proliferate and differentiate
antagonist Noggin we found a previously not described into immature neurons. Although the number of mature
large mesenchymal Noggin source that sharply defines a neurons and the in vivo rate of proliferation were not altered,
BMP free GnRH and VNO neurogenic border. In Fgf8 the proliferative response to the olfactotoxicant satratoxin
hypomorphs BMP4 and Noggin expression were found to G and olfactory bulb ablation injury was compromised in
be altered, with Noggin no longer defining the GnRH and the olfactory epithelium of IP3R3−/− mice. The reductions in
both NPY release and number of progenitor cells in IP3R3−/− #P74 POSTER SESSION II: OLFACTION
mice point to a role of the IP3R3 in tissue homeostasis and DEVELOPMENT; TASTE CNS; NEUROIMAGING;
neuroregeneration. Collectively, these data suggest that OLFACTION CNS
IP3R3 expressing microvillous cells are actively responsive
to injury and promote recovery. Acknowledgements: Inactivation of the Interoceptive Insula Suppresses
Supported by NIH DC006897 (CCH), MSU institutional Chemosensory Cue Reactivity to Ethanol Following Chronic
funds (CCH), NIEHS T32 ES007255 (CRH), NINDS Ethanol Exposure
T32 NS044928 (AEP and TRI) and Swiss Fellowship for Norma Castro, Emily Driver, Jason Dudley, Jessica Godfrey,
Advanced Researchers PA 00P3_131493 (SH). Brian Feretic, Carlo Quintanilla, Susan M. Brasser
San Diego State University/Psychology San Diego, CA, USA
DEVELOPMENT; TASTE CNS; NEUROIMAGING; Recent findings indicate that the insular cortex is critically
OLFACTION CNS involved in addictive behavior to multiple drugs of abuse by
regulating an organism’s responsiveness to drug-associated
PACAP increases [Ca2+] in neonatal OB via direct and indirect sensory cues. Damage to the insula in addicted smokers
mechanisms results in a disruption of nicotine addiction, and inactivation
Mavis A Irwin1, Mary T Lucero2 of the insula in rodents disrupts conditioned preference
for environments previously paired with amphetamine or
University of Utah Salt Lake City, UT, USA, 2American
1
nicotine. Imaging studies have demonstrated activation
University of the Caribbean Cupecoy, Netherlands Antilles in the insula during drug craving and exposure to drug-
Our lab has been studying the pleiotropic peptide named related cues, including the taste of alcohol in heavy drinkers.
Pituitary Adenylate Cyclase Activating Peptide (PACAP) The present study measured chemosensory responses to
in the olfactory epithelium1 of rodents. The physiologi- ethanol in chronically ethanol- exposed or naive rats under
cal effects of PACAP in the olfactory bulb (OB) are still conditions of pharmacological silencing of the visceral
unknown. Neonatal OB is enriched with both PACAP and insula to examine the role of this brain region in mediating
its G-protein coupled receptor PAC1R. Without PACAP, responses to ethanol-associated sensory cues. Rats were
neonates often die before weaning, suggesting that PACAP initially exposed to either a 20% ethanol intermittent access
is required for normal development. Previously, we showed paradigm or were given access only to water. Following
that PACAP led to an oscillating increase in [Ca2+] in OB implantation of intracranial cannulae, rats from each
neurons. To address whether the PACAP-induced responses exposure condition were tested for brief-access lick responses
are direct or indirect, we used cocktails of antagonists for to ethanol (0–40%) after receiving bilateral insula infusions
the GABA receptors (GABAR) and/ or glutamate recep- of saline or muscimol. Alcohol-experienced rats displayed
tors (GlutR) in the presence and absence of PACAP. We a concentration-dependent increase in chemosensory avidity
performed confocal Ca2+ imaging on live slices from P2-P5 for ethanol compared to alcohol-naive rats, evidenced
mice loaded with the Ca2+ indicator dye Fluo-4 AM. The by elevated lick responses and trial sampling frequency
optimal dose of PACAP was empirically determined to be particularly at higher ethanol concentrations (15–40%).
40 nM and was used in all experiments. Combined block of Inactivation of the insula eliminated this concentration-
GABAR and GlutR yielded a 66% decrease in numbers of dependent response in chronically exposed animals, but
PACAP responsive cells. Blocking just GlutR resulted in a did not modify orosensory responses to ethanol in alcohol-
similar reduction, suggesting that glutamate mediates the naive rats. These data support insular cortex involvement
majority of the indirect effects. Interestingly, blocking only in mediating responsiveness to conditioned alcohol
the GABAR resulted in block of GABA-induced initial Ca2+ chemosensory cues following chronic association of ethanol’s
response on immature cells. However, the majority of these taste and post-absorptive effects. Acknowledgements:
cells showed the post-PACAP oscillation. Our data sug- Support Contributed By: NIH AA015741
gest 1) about 1/3 of the PACAP-responsive cells have direct
PAC1R activity. 2) PACAP promotes glutamate release
which in turn activates 2/3 of the PACAP-responsive cells. #P75 POSTER SESSION II: OLFACTION
3) GlutR may have a role in the post-PACAP [Ca2+] oscil- DEVELOPMENT; TASTE CNS; NEUROIMAGING;
lation. 4) GABA is also released by PACAP from PAC1R- OLFACTION CNS
rich GABAergic cells. In conclusion, we find that PACAP Growth patterns of sensory neuron axon terminals in the
has both direct and indirect effects on neonatal OB neurons developing olfactory bulb
and may promote glutamate and GABA release in early
development. Acknowledgements: NIH 1F31DC011686-02 Ivan Manzini1,2, Thomas Hassenlklöver1,2
Blackman Trust Fund 00253 6000 16917 University of Utah 1
University of Göttingen, Department of Neurophysiology
Graduate Research Fellowship 2011 and Cellular Biophysics Göttingen, Germany, 2University of
Göttingen, DFG Cluster of Excellence “Nanoscale Microscopy considerable neurogenesis throughout life. In the mouse
and Molecular Physiology of the Brain” (CNMPB) Göttingen, olfactory epithelium, each primary olfactory sensory neuron
Germany (OSN) stably expresses a single odorant receptor (OR) type
The developing, but also the mature vertebrate olfactory sys- out of a repertoire of ~1200. All OSNs with the same OR
tem is a site of ongoing neurogenesis. Olfactory stem cells identity are distributed within one of the few broadly-
continuously generate new sensory neurons which extend defined zones. However, it remains elusive whether such
axons into the olfactory bulb where they face the challenge to OR expression patterns are shaped by sensory stimulation
integrate into an existing neuronal circuitry. Synaptic contacts and/or neuronal activity. Here we addressed this question
to second-order neurons are formed in distinct target regions, by investigating OR gene or protein expression in two
so-called glomeruli. In rodents, sensory neurons normally surgically- or genetically-modified mouse models. Using in
project only into one specific glomerulus of the olfactory bulb. situ hybridization, we examined the expression patterns of
We investigated the growth patterns of sensory neuron axons 15 selected OR genes in mice which underwent neonatal,
in the developing olfactory system of the aquatic amphibian unilateral naris closure. After four-week occlusion, the
Xenopus laevis. To address the question how connectivity is expression level in the closed side was significantly lower (for
reshaped during olfactory system maturation a range of lar- four ORs), similar (for three ORs) or significantly higher (for
val stages and young postmetamorphic animals were included eight ORs) than that in the open side. In addition, using a
in the experiments. Fluophore-coupled dextrans or plasmid specific OR antibody, we demonstrated that this OR protein
DNA, encoding for fluorescent proteins, were introduced into was upregulated in the closed side but downregulated in
sensory neurons via electroporation. The main sensory projec- the open side. Furthermore, we examined the expression
tion fields within the main- and accessory olfactory bulb were patterns of individual OR genes in transgenic mice in which
visualized by electroporation of the whole olfactory organ. olfactory marker protein (OMP) drives overexpression of
During metamorphosis the main olfactory system is com- the inward rectifying potassium channel (Kir2.1) in most
pletely reorganized, whereas the sensory neurons of the acces- mature OSNs to reduce their neuronal activity. The cell
sory olfactory system are maintained. The axonal branching density for most OR genes (six out of seven tested) was
patterns of sensory neurons, originating from both the vome- significantly reduced compared to wild-type controls. The
ronasal and main olfactory epithelium, were investigated by results suggest that sensory inputs have differential influence
sparse staining of sensory neurons. Synaptic connections on OSNs expressing different ORs and that neuronal
were clearly visible as tufted axonal endings. Most sensory activity is critical for survival of OSNs. Acknowledgements:
neurons showed a branched axonal pattern before terminat- Supported by grants from the NIDCD/NIH DC006213 and
ing in tufted arborizations inside glomeruli. Surprisingly, a DC011554.
high percentage of cells terminated in multiple and not single
glomerulus-like structures. This pattern was comparable in
sensory neurons originating from both the vomeronasal and #P77 POSTER SESSION II: OLFACTION
the main olfactory organ. Acknowledgements: Supported DEVELOPMENT; TASTE CNS; NEUROIMAGING;
by DFG Cluster of Excellence “Nanoscale Microscopy and OLFACTION CNS
Molecular Physiology of the Brain” (CNMPB) to I.M. and Optogenetic Investigation of GABAergic Circuitries in the
DFG Schwerpunktprogramm 1392 to I.M. Rostral Nucleus of the Solitary Tract
James A. Corson, Robert M. Bradley
#P76 POSTER SESSION II: OLFACTION University of Michigan/ Biologic and Materials Sciences Ann
DEVELOPMENT; TASTE CNS; NEUROIMAGING; Arbor, MI, USA
OLFACTION CNS
The rostral nucleus of the solitary tract (rNTS) is the first
Activity-Dependent Expression of Odorant Receptors in the central target of primary gustatory nerve fibers and as
Mouse Olfactory Epithelium such plays an essential role in the processing and coding
Shaohua Zhao1,2, Huikai Tian1, Rosemary Lewis1, Limei Ma3, of peripheral taste sensory information. The intrinsic cir-
Ying Yuan1, Congrong R Yu3, Minghong Ma1 cuitry within rNTS is likely integral in shaping the incom-
ing information into both ascending and descending efferent
1
Department of Neuroscience, University of Pennsylvania
signals. Substantial subpopulations of interneurons in the
School of Medicine Philadelphia, PA, USA, 2Department of
rNTS are GABAergic and thus contribute to the generation
Geriatric Cardiology, Qilu Hospital of Shandong University
of hyperpolarization-activated changes in repetitive firing
Jinan, China, 3Stowers Institute for Medical Research Kansas
patterns in projection neurons. Despite this importance in
City, MT, USA
shaping rNTS gustatory-evoked signaling, the organization
Sensory experience plays critical roles in development and of rNTS GABAergic circuits is unknown. To investigate
maintenance of the olfactory system, which undergoes the organization of GABAergic innervation onto identified
populations of neurons, we used a mouse model in which additional cell was not responsive to lingual taste stimuli but
channelrhodopsin was expressed under the control of the was inhibited by gastric tastant delivery of MSG, HCl and
vesicular GABA transporter. GABAergic interneurons were quinine. Collectively, these data suggest that information
activated in an in vitro slice preparation with 473 nm laser from both lingual and post-lingual chemoreceptors
illumination merged into the optic train of the microscope. converge onto NTS cells, suggesting a role for post-lingual
Focused laser illumination produced consistent saturated chemoreceptors in modulation of ingestive behavior on a
photocurrents in GABAergic neurons with high temporal short timescale. Acknowledgements: Supported by NIDCD
and spatial resolution. While recording inhibitory postsyn- grant RO1DC006914 to PMD.
aptic currents in either GABAergic or non-GABAergic neu-
rons, the laser spot was systematically scanned over discrete
portions of the rNTS to map out the GABAergic innerva- #P79 POSTER SESSION II: OLFACTION
tion onto the recorded neuron. Neurons received inhibitory DEVELOPMENT; TASTE CNS; NEUROIMAGING;
innervation from wide expanses of rNTS, often with focal OLFACTION CNS
spots of strong inhibition located in areas not immediately
adjacent to the recorded neuron. This suggests that in addi- Synaptic Properties of the Basolateral Amygdala Projection
tion to a low level of global inhibition, there are also specific to Gustatory Cortex
subregions of rNTS that are able to strongly hyperpolarize Melissa Haley1,2, Alfredo Fontanini1,2, Arianna Maffei1,2
individual neurons and possibly induce alterations in repeti-
Department of Neurobiology and Behavior Stony Brook,
1
tive discharge patterns. Acknowledgements: T32DC000011,
NY, USA, 2Program in Neuroscience Stony Brook, NY, USA
RO1DC000288
The gustatory cortex (GC) receives a dense and anatomi-
cally well documented projection from the basolateral
#P78 POSTER SESSION II: OLFACTION nucleus of the amygdala (BLA). This input is thought to
DEVELOPMENT; TASTE CNS; NEUROIMAGING; convey affective and anticipatory information regard-
OLFACTION CNS ing taste. Indeed inactivation of BLA results in dramatic
changes in the way taste and anticipatory cues are processed
Sensory Afferents from the Stomach of the Rat Converge in GC. Although the behavioral and functional significance
onto Taste-responsive Neurons in the Rat Brainstem of this projection has been the focus of extensive investiga-
Alexander J. Denman-Brice, Patricia M. Di Lorenzo tion in awake and anesthetized animals, the synaptic proper-
ties and organization of these inputs are unknown. To study
Binghamton University/Psychology Binghamton, NY, USA
the BLA to GC synapse, viral vectors carrying a construct
Recent descriptions of taste receptors in the gut have set for ChannelRhodopsin2 were injected in the BLA. After 2
the stage for the idea that taste stimuli continue to provide weeks, whole-cell recordings in dysgranular and agranular
information even when they are no longer in the mouth. GC were performed in combination with photoactivation
For example, intraduodenal infusions of bitter tastants can of BLA terminal fields. BLA afferents were found to target
modify eating behavior within a single meal. Given that both excitatory and inhibitory neurons in all layers of GC.
both taste and post- lingual chemosensory feedback may Across all layers, approximately 60% of regular- spiking
be important for modifying eating behavior on a relatively (RS), fast-spiking (FS), and low-threshold spiking (LTS)
short timescale, it is possible that chemosensory afferents neurons responded to light-activation of BLA afferents.
from the gut may converge onto the same relay nuclei as RS cells had a longer rise time than FS cells (p=.016) and
taste information in the brainstem. We investigated the a longer decay time than FS cells (p=.0004). In addition,
responsiveness of single neurons in the rat brainstem to differences could be seen in the synaptic properties of BLA
tongue and gastric taste stimulation. Initially, rats were inputs onto neurons in superficial and deep layers of GC.
anesthetized with urethane and prepared for recording from Layer 2/3 RS cells had larger current amplitudes than layer
the brainstem. A length of polyethylene tubing was threaded 5/6 cells (p=.03), and there was a significant difference in
down the rat’s esophagus to the stomach for delivery of the percentage of FS cells that responded to stimulation in
tastants. A tungsten microelectrode was then placed in the layer 2/3 (75%) and layer 5/6 (25%). These data suggest that
nucleus of the solitary tract (NTS) and taste-responsive cells BLA inputs in GC have cell type specific and layer specific
were isolated. Preliminary data from 18 taste-responsive properties. The combination of feed-forward inhibition and
cells show that some (n=8) NTS cells change their firing rate excitation likely serves to shape the temporal dynamics of
in response to infusion of small amounts (0.4 ml) of taste taste responses and to enhance the representation of behav-
stimuli (0.1 M NaCl, 0.01 M HCl, 0.01 M quinine, 0.5 M iorally salient stimuli. Acknowledgements: National Eye
sucrose and 0.1 M MSG) into the stomach. Gastric responses Institute Grant #R01-EY019885-S1 and National Institute
were most frequently found to NaCl and MSG; no excitatory on Deafness and Other Communication Disorders Grant
responses were found to HCl infused into the stomach. An #R01-DC010389
#P80 POSTER SESSION II: OLFACTION (100%/0%, 90%/10%, 70%/30%, 50%/50%, etc.). We tested for
DEVELOPMENT; TASTE CNS; NEUROIMAGING; three different hypothetical response patterns: 1) Responses
OLFACTION CNS varying as a function of sucrose concentration (the
monotonic pattern); 2) Responses increasing or decreasing as
Cortical modulation of taste-related orofacial behaviors a function of degree of mixture of the stimulus (the mixture
Jennifer X. Li, Donald B. Katz pattern); and 3) Responses that change abruptly from being
similar to one pure taste to being similar the other (the
Brandeis University Waltham, MA, USA
categorical pattern). Our results demonstrate the presence
Upon receiving a taste stimulus, primates and rodents produce of both monotonic and mixture patterns within responses of
a sequence of rhythmic orofacial movements, also known as GC neurons. Specifically, further analysis (that included the
taste reactivity (TR), specific elements of which reflect the presentation of 50 mM sucrose and citric acid) made it clear
hedonic quality (i. e., the palatability) of the stimulus. Aversive that mixture suppression reliably precedes a palatability-
stimuli, such as quinine, elicit bouts of gapes, movements related pattern, and that the same phenomenon characterizes
that serve to eject the stimulus from the mouth. The produc- responses to sucrose/NaCl mixtures. The temporal dynamics
tion of gapes indicates aversiveness, and the latency to gape of the emergence of the palatability-related pattern parallel
is inversely related to the degree of aversiveness (Travers and the temporal dynamics of the emergence of preference
Norgren, 1986). While the motor circuits necessary for taste- behavior for the same mixtures, as measured by a brief access
related orofacial movements are contained within a brainstem test. We saw no evidence of categorical coding.
network (Grill & Norgren), in the intact animal, this network
is modulated by feedback from higher-order forebrain struc-
tures. Relatively little is known about forebrain structures’ roles #P82 POSTER SESSION II: OLFACTION
in the selection and production of TR, however. We set out to DEVELOPMENT; TASTE CNS; NEUROIMAGING;
examine the relationship between TR and neural responses in OLFACTION CNS
primary gustatory cortex (GC) by performing paired record-
ings of single unit activity and jaw electromyography (EMG) Conditioned Taste Aversion Does Not Require Cortical
in awake rats presented with strong (0.3 M) and weak (0.03 M) mRNA Synthesis
sucrose, and strong (0.001 M) and weak (0.0001 M) quinine, Abigail A Russo1, Yasmin U Marrero2, Donald B Katz1,2,3
via intra-oral cannulae. Comparisons of the time courses of
neural and EMG responses revealed that palatability-related
1
Brandeis University Department of Psychology Waltham,
signals in GC preceded those in EMG by ~250 ms, suggest- MA, USA, 2Brandeis University Program of Neuroscience
ing that GC could drive palatability-related orofacial move- Waltham, MA, USA, 3Volen Center for Complex Systems,
ments. Preliminary analyses further demonstrated that the Brandeis University Waltham, MA, USA
spiking activity of individual GC neurons was correlated with Although it has been well established that the gustatory cor-
the latency of quinine-induced gape bouts. Our results suggest tex (GC) plays a significant role in the consolidation of taste
that forebrain activity may influence certain aspects of TR, memory, the precise physiological mechanisms by which this
such as the initiation of palatability-related orofacial move- takes place are not fully understood. Notably, taste memory
ments. Acknowledgements: DC007703 acquisition is traditionally viewed as dependent on cortical
protein synthesis (Dudai et al. 2004), but it is unclear whether
#P81 POSTER SESSION II: OLFACTION the learning process requires cortical mRNA transcription.
DEVELOPMENT; TASTE CNS; NEUROIMAGING; Here, we investigated this possibility using actinomycin D
OLFACTION CNS (Act-D), an mRNA synthesis inhibitor that has been shown
to impair contextual fear conditioning when infused into the
Neural dynamics in response to binary taste mixtures amygdala (Parsons et al 2006). Act-D was microinfused via
Joost X Maier, Donald B Katz cannulae implanted into the GC (1.4 mm anterior to Bregma,
5.0 mm lateral, 4.5 mm ventral) of awake rats. Immediately
after infusion, a conditioned taste aversion protocol was per-
In natural environments, taste signals an animal encounters formed during which the rats were exposed to a taste paired
typically consist of complex mixtures of tastants. Although with malaise. Our results indicated that rats form taste aver-
a great deal is known about how the taste system processes sions even when mRNA synthesis in GC is blocked. These
single tastes presented in isolation, not much is know results suggest that memory consolidation is in part inde-
about how brain integrates different taste signals presented pendent of mRNA synthesis in the gustatory cortex. It is
simultaneously. Here we probed single neurons in primary likely that subcortical production of mRNA, presumably in
gustatory cortex (GC) for responsiveness to binary taste the amygdala, is sufficient to support cortical protein syn-
mixtures. Stimuli consisted of sucrose and citric acid and thesis and establish taste memory. Acknowledgements: R01
sucrose and sodium chloride mixed in different ratios DC-006666/DC/NIDCD NIH HHS/ United States
#P83 POSTER SESSION II: OLFACTION Biologic & Materials Sciences, School of Dentistry University
DEVELOPMENT; TASTE CNS; NEUROIMAGING; of Michigan, MI, USA
OLFACTION CNS All chemosensitive information derived from stimulation
Nutritive value, not taste, is necessary for flavor of taste receptors in the oro-pharynx relays via the rNST
preferences in mice in the brain stem. Neurons of the rNST have been defined
using anatomical and histochemical criteria but recently
Jennifer M Stratford we have also shown that many rNST neurons possess pri-
Rocky Mountain Taste & Smell Center, Department of mary cilia, small organelles extending from the cell surface.
Cell and Developmental Biology, Neuroscience Program, Primary cilia have been shown to play an important role dur-
University Colorado School of Medicine Aurora, CO, USA ing development and are involved in cell signaling pathways.
The importance of cilia in development is evident in various
The preference for food is dependent primarily upon the
developmental brain disorders, or ciliopathies, resulting from
interplay between oral and post-oral factors. The relative
disrupted cilia function. We studied the location of primary
contribution of these two systems to food intake is not fully
cilia in rNST cells in postnatal rat because they may play a
explored. Mice that lack the ability to taste, by genetic deletion
role in signaling during taste processing. A number of mark-
of the P2X2/P2X3 purinergic receptor subunits (P2X-KO),
ers have been used to identify primary cilia. Two widely used
can form a preference for monosodium glutamate (MSG)
markers are ACIII, part of a cAMP pathway, and Arl13b,
using only post-ingestive cues. The neural mechanisms that
part of a cGMP signaling pathway. Previously we reported
underlie this ability remain unknown but likely involve
that ACIII-labeled primary cilia are present in about half
viscerosensory detection of the nutritive qualities of MSG.
of rNST neurons. Somatostatin receptor 3 (SSTR3) and
Thus, the current study assessed if P2X-KO mice can form
melanin-concentrating hormone receptor 1(Mch1R) co-
a preference for the calorie-free sweetener, SC45647 (SC),
localize with the ACIII-labeled cilia. Interestingly, though,
which, like MSG, is appetitive to WT animals. WT and
Arl13b and ACIII-labeled cilia are found on different rNST
P2X-KO mice were given training sessions with a flavor alone
cells. Neither SSTR3 nor Mch1R co-localizes with Arl13b.
(e.g. cherry) or with a different flavor (e.g. grape) mixed with
In addition, recently we detected a primary cilium in almost
0.05 mM SC. Then all animals were given 2-bottle preference
every rNST astrocyte using an Arl13b antibody, but not
tests with both flavors without SC. During training, WT
ACIII, further demonstrating differences in rNST primary
animals drank more SC than did P2X-KO mice, suggesting
cilia. Since ACIII couples with a cAMP pathway and Arl13b
that WT, but not P2X-KO mice, can taste SC. However,
couples with a cGMP signaling pathway it is possible that
neither WT nor P2X-KO animals preferred the flavor that
the cell types that express different primary cilia play dif-
was previously paired with SC in flavor alone preference tests.
ferent roles in rNST function. Acknowledgements: NIH
SC-evoked brain activation was measured by expression of
NIDCD Grant DC000288
the immediate early gene c-Fos (cFLI) in the nuc. solitary
tract (nTS)- the primary taste/viscerosensory nucleus. As
previously reported for MSG stimulation, SC- induced #P85 POSTER SESSION II: OLFACTION
cFLI in gustatory (rostral) nTS was less in P2X-KO animals DEVELOPMENT; TASTE CNS; NEUROIMAGING;
compared to WT controls. In viscerosensory (caudal) nTS, OLFACTION CNS
SC-induced cFLI did not differ between WT and P2X- KO
mice. Further, within caudal nTS, SC was less effective than Taste responses change over consecutive days in single cells
MSG in evoking cFLI in both lines. Together, these results in the rat brainstem recorded in the awake animal
suggest that nutritive content, not taste, is necessary to drive Michael S. Weiss1, Andrew M. Fooden1, Jonathan D. Victor2,
food preferences and that this information is represented Patricia M. Di Lorenzo1
in the caudal nTS. Acknowledgements: Supported by NIH 1
Binghamton Univeristy Dept. of Psychology Binghamton,
grants to JMS and Thomas E. Finger (U. of Colorado
NY, USA, 2Weill Cornell Medical College Dept. of Neurology
Denver Medical School, Rocky Mountain Taste and Smell
and Neuroscience New York, NY, USA
Center, Aurora, CO).
Theories of taste coding have relied on recordings from
single cells in a single session; i.e. snapshots of activity. In
#P84 POSTER SESSION II: OLFACTION contrast, there is evidence that taste responses can change
DEVELOPMENT; TASTE CNS; NEUROIMAGING; over days (e.g. in chorda tympani fibers, Shimatani et al.,
OLFACTION CNS Physiol. & Behav., 80(2–3), 309–15, 2003). Here, we present
data showing that taste responses in individual cells in the
Distinct groups of cilia in rat rostral nucleus of the solitary
nucleus of the solitary tract (NTS) and parabrachial nucleus
tract (rNST) labeled with ACIII and Arl13b
of the pons (PbN) vary significantly across consecutive days.
Min Wang, Charlotte C. Mistretta, Robert M. Bradley Rats were surgically implanted with a chronic microwire
assembly into the NTS or PbN, allowed to recover, and water of activation of ChAT-expressing cells in the EPL in both
deprived. Rats had free access to a lick spout that delivered exposure groups when compared to activation in control
taste stimuli or water while cellular activity was recorded. mice, suggesting that this population may serve a role in the
Thus far, in the NTS, 8 animals yielded multi-day recordings processing of social odor cues. Thus, we have identified a
(range = 2–5 d; median = 2 d); in the PbN, 5 animals significant cholinergic interneuron population in the AOB
yielded multi-day recordings (range = 2–7 d; median = 2.5 that varies significantly in the anterior and posterior regions.
d). To determine whether the recordings on successive days Thus, our data supports the idea that the anterior and pos-
were likely to represent recordings of the same neuron, we terior AOB process sensory information differently and sug-
examined the similarity of the recorded waveform templates. gests that this cholinergic interneuron population may serve
For 76% of multi-day NTS recordings and 30% of multi-day to process olfactory information in a region specific man-
PbN recordings, waveforms were highly similar (waveform ner. Acknowledgements: Supported by research grants NIH/
template correlation > 0.99). As a control, this degree of NIDCD 009269, 012831 and ARRA administrative supple-
similarity was rare (1.3% of pairs in NTS, <1% of pairs ment to WL
in PbN) when the waveforms were from known-different
neurons, recorded on separate microwires. Thus, it is likely
most of the recordings across days represent recordings of the #P87 POSTER SESSION II: OLFACTION
same neuron. Analyses of these putative same-cell multi-day DEVELOPMENT; TASTE CNS; NEUROIMAGING;
recordings showed that responses to individual tastants both OLFACTION CNS
appeared and disappeared across days, resulting in shifts in
tuning. These data imply that theories of taste coding need Suppression of Association Synapses in Piriform Cortex
to incorporate the dynamic nature of taste response profiles. During Post-Training Sleep Impairs Odor Memory Selectivity
Acknowledgements: NIDCD R01-DC006914 Dylan C Barnes1,2, Donald A Wilson1,2,3
1
Graduate Center CUNY New York City, NY, USA, 2Nathan
Kline Institute Orangeburg, NY, USA, 3NYU Langone Medical
#P86 POSTER SESSION II: OLFACTION Center New York City, NY, USA
OLFACTION CNS Slow wave sleep (SWS) is characterized by slow-wave
oscillations in neocortex, as well as sharp waves (SPW) in
Characterization of Cholinergic Interneuron Populations in both the hippocampus and piriform cortex (PCX). Neural
the Accessory Olfactory Bulb activity during SWS is hypothesized to contribute to memory
Sarah E Ashby, Kurt Krosnowski, Weihong Lin consolidation through “replay” of waking activity patterns.
For example, we have demonstrated that imposed replay
University of Maryland, Baltimore County Baltimore,
of odor- evoked activity in the olfactory system during
MD, USA
SWS enhances subsequent memory of that odor. Neurons
The accessory olfactory bulb (AOB) contains diverse pop- co-activated by an odor are hypothesized to become
ulations of intrinsic interneurons that play a key role in linked into a cohesive ensemble through strengthening
information processing, including choline acetyltransferase of association synapses. Replay of odor evoked ensemble
(ChAT)- expressing interneurons, which have not previously activity during SWS may help strengthen these connections
been identified. We detected these cholinergic interneurons and improve memory and memory acuity. Here, we tested
via green fluorescent protein (GFP) signal in ChAT(BAC)- the hypothesis of that association fiber activity during SWS
eGFP mice, which allows visualization and quantification facilitates replay and memory of recently learned odors
of the interneuron cell bodies throughout the bulbar layers by infusing baclofen (or saline) into the PCX during post-
of the AOB (Krosnowski et al, 2012). We found significant training sleep. Baclofen is a GABA-B receptor agonist
differences in the number of cholinergic interneurons in the that has been shown to selectively depress association fiber
anterior and posterior glomerular layer (GL) and the exter- synapses. Rats were chronically implanted with bilateral
nal plexiform layer (EPL), with the highest numbers of cho- cannulae and a recording electrode in the anterior PCX.
linergic interneurons in the anterior GL and posterior EPL. After recovery, rats were differentially conditioned with CS+
In the posterior EPL, we also noted a high density of GFP+ odor/footshock and CS− odor stimuli. During the 4 hours
cells forming a ring around the outer edges of the layer, thus immediately post-training, animals were placed in a sleeping
creating a heavy GFP+ population along the border between chamber and bilaterally infused with either baclofen or saline.
the anterior and posterior AOB. We then examined the pos- Local field potential and EMG activity were recorded during
sible role of ChAT interneurons in the AOB using adult male conditioning, post-training sleep, and test periods. On test
mice exposed to either bedding from a mated pair or to a day, 24 hours following conditioning, freezing responses to
male aggressor mouse. We monitored the activity marker, the CS+, CS− and other odors were examined. Preliminary
c-fos, in these regions and found significantly higher levels behavioral results suggest that post-training PCX baclofen
infusions do not impair memory for the CS+ but reduce 4

Northwestern University/Department of Neurobiology
odor acuity/enhance generalization of the odor-fear Evanston, IL, USA, 5HHMI Janelia Farm Research Campus/
response. Acknowledgements: F31-DC012284 to D.C.B. and Visiting Scientist Program Ashburn, VA, USA, 6The University
R01-DC003906 from the NIDCD to D.A.W. of Texas Medical School/Department of Neurobiology and
Anatomy Houston, TX, USA
#P88 POSTER SESSION II: OLFACTION Olfactory bulb glomeruli organize and relay sensory infor-
mation that arrives from the nose. Odors typically evoke
activity across many glomeruli, making it difficult to study
OLFACTION CNS
their individual role in olfactory processing. To overcome
Cholinergic modulation of glomerular odor sensitivity in this difficulty we employed transgenic mice in which chan-
the olfactory bulb nelrhodopsin-2 is selectively expressed in genetically iden-
Mounir Bendahmane, M Cameron Ogg, Max L Fletcher tified olfactory sensory neurons; these neurons project
their axons to a defined glomerulus and can be activated
University of Tennessee Health Science Center Memphis, via pulsed illumination of the olfactory epithelium with a
TN, USA 447nm laser. Through in vivo two-photon calcium imaging,
In the olfactory system, many studies have shown that cho- we monitored optically evoked neural activity in the glomer-
linergic input to the olfactory bulb is not only involved in ulus and nearby juxtaglomerular neurons. Laser pulses reli-
learning and memory but also detection and discrimination. ably activated the glomerulus and evoked calcium responses
In this study we used calcium imaging to explore the cholin- in juxtaglomerular neuron pools of various sizes. Changing
ergic effect on OB postsynaptic glomerular odor responses. laser pulse width and timing altered the strength, appear-
Using mice expressing GCaMP2 in M/T cells, we studied ance and duration of glomerular and cellular responses.
the modulation of dorsal surface glomerular odor concen- We also identified juxtaglomerular cells that responded to
tration-response curves via HDB (horizontal limb of the optical activation with decreased intracellular calcium;
diagonal band of Broca) stimulation or OB cholinergic phar- unlike activated cells, these inhibited cells did not cluster
macological manipulation. Overall, we find that increased next to the optically activated glomerulus but were spread
cholinergic OB activation through HDB stimulation or cho- throughout surrounding areas. We are now investigating
linergic-uptake blocker application increases the sensitivity how different neuronal responses types relate to the molecu-
of individual glomerular odor responses by shifting the odor lar phenotype of the juxtaglomerular cells. To accomplish
concentration-response curve to the left and decreasing the this we generated hybrid mice, which express GAD67-GFP
EC50 by up to one log unit in odor concentration. This effect and/or GAD65- GFP in olfactory bulb interneurons along
was observed for all glomeruli tested regardless of baseline with the selectively expressed channelrhodopsin-2. In com-
odor sensitivity or odorant used. OB application of a mus- bination, our data provide a uniquely detailed analysis of
carinic antagonist completely blocks these shifts, suggesting the neuronal network that comprises a single olfactory glo-
that the increased sensitivity observed is primarily driven merulus. Acknowledgements: Supported by NIH grants
by muscarinic activation. We are now exploring the cholin- U24NS057631 and R01DC005259 and by the National
ergic effects on individual OB cell types using two-photon Research Foundation of Korea World class Institute Grant
microscopy to further address these effects at the single cell WCI 2009-003.
level. Acknowledgements: NIH R03 DC009853 and the Pew
Scholars Program in the Biomedical Sciences.
#P89 POSTER SESSION II: OLFACTION OLFACTION CNS
DEVELOPMENT; TASTE CNS; NEUROIMAGING; Top-down modulation of olfactory bulb output by the
OLFACTION CNS midbrain serotonergic system.
In Vivo Optophysiological Analysis of the Glomerular Unit Daniela Brunert, Matt Wachowiak
Response in Mice
Brain Institute and Department of Physiology Salt Lake City,
Oliver R. Braubach1,2,3, Tuce Tombaz1, Masoud UT, USA
Allahverdizadeh1, Thomas Bozza4,5, Lawrence B. Cohen1,2,3,
The olfactory bulb (OB) receives input from multiple top-
Ryota Homma2,3,6
down neuromodulatory systems. Serotonergic inputs from
1
Korea Institute of Science and Technology/Center for the raphe innervate all OB layers and can presynaptically
Functional Connectomics Seoul, Korea, 2Marine Biological modulate sensory input gain. However, the effects of
Laboratory Woods Hole, MA, USA, 3Yale School of serotonergic modulation on OB circuitry and output in vivo
Medicine/Department of Physiology New Haven, CT, USA, remain unclear. Here, we used Cre-expressing mouse strains
to express the calcium indicator GCaMP in periglomerular cortex. Furthermore, in the LO injected mouse, we found
(PG) cells (using GAD2-cre mice) and in mitral/tufted cells labeled cells in agranular insular and mediodorsal nucleus
(MTCs) (Cdhr1-cre mice) in order to visualize how raphe of thalamus. In the AI injected mice, a similar labeling pat-
stimulation alters resting and sensory- evoked excitation of tern was seen in endopiriform nucleus with sparsely labeled
these two neuron populations. In GAD2- cre mice, brief (1–4 cells in ventral medial anterior piriform cortex. In both
s) electrical stimulation of raphe elicited a slow increase in experiments, we found no labeled cells in the dorsal anterior
baseline fluorescence that outlasted stimulation by several piriform cortex or posterior piriform cortex. Together the
seconds, as well as a several-fold increase in the amplitude data suggest that piriform cortex may be composed of finer
of inhalation-evoked transients (mean increase, 650 ± 247%). anatomical subdivisions that project to prefrontal areas.
Stimulation of raphe also increased odorant-evoked Whether these subdivisions perform specific functional roles
response amplitudes. Raphe stimulation effects were blocked remains to be determined.
by the 5-HT2A/C antagonist cinanserin applied locally
to the OB. These results suggest that serotonergic inputs
to OB transiently increase the baseline excitability of PG #P92 POSTER SESSION II: OLFACTION
cells as well as their responses to sensory input. In contrast, DEVELOPMENT; TASTE CNS; NEUROIMAGING;
MTCs in Cdhr1-cre mice showed only weak increases OLFACTION CNS
in baseline fluorescence upon raphe stimulation onset
followed by a more pronounced increase after stimulation Neuromodulatory regulation of learning within
offset in some animals. Surprisingly, raphe stimulation did olfactory bulb
not alter inhalation- or odorant-evoked responses in these Thomas A Cleland
neurons. Together, these results demonstrate a differential
Dept. Psychology, Cornell University Ithaca, NY, USA
effect of serotonergic modulation on OB cell types in vivo
and serve as a starting point for further dissection of the Intrinsic plasticity within olfactory bulb (OB) circuitry
circuit mechanisms underlying the top-down modulation of modifies odor generalization gradients based on experience
early olfactory processing as a function of behavioral state. so as to dynamically construct essentially categorical odor
Acknowledgements: Funded by NIDCD DC010915 representations. This plasticity also is regulated and perhaps
gated by neuromodulatory state, and can be manipulated
by pharmacological infusions into OB. Cholinergic
#P91 POSTER SESSION II: OLFACTION inputs to the OB acutely regulate the breadth of odor
DEVELOPMENT; TASTE CNS; NEUROIMAGING; generalization, though the nicotinic component, primarily
OLFACTION CNS localized in the glomerular layer, dominates this acute
effect. In contrast, muscarinic receptors are predominantly
Neuronal connections from piriform cortex to prefrontal expressed in the external plexiform layer (EPL), which has
cortical areas of mice been increasingly associated with mechanisms of intrinsic
Chien-Fu F Chen, Natasha Kharas learning within OB. We therefore investigated the role that
muscarinic modulation of OB circuitry plays in olfactory
Stuart Firestein Columbia University/Biological Sciences New
associative learning. We found that intrabulbar infusion of
York, NY, USA
scopolamine impaired olfactory learning when delivered
Piriform cortex, the main olfactory processing area, has been between training and testing, or when delivered prior to
shown to have projections to prefrontal cortex providing training in studies imposing a similar 45-minute training-
olfactory input and receive projections from prefrontal cor- testing latency, but not when testing followed training
tex as a potential downstream modulatory pathway. While immediately or with a four- minute latency. (Dihydrokainate
recent data establish the projections from the prefrontal infusion into OB was used to prevent the retrograde
areas to piriform, the projections from piriform to prefrontal amnestic effect of isoflurane anesthesia). This pattern of
areas remain less understood. To investigate these connec- results indicates that intact muscarinic responsivity within
tions, we utilized retrograde tracing and confocal micros- OB is important for the maintenance of an intact odor
copy. We injected the retrograde tracer, cholera toxin subunit memory over this delay period. In contrast, intact alpha-1
b (CTb), via iontophoretic injection in the prefrontal areas noradrenergic responsivity in OB appears permissive for
of lateral orbitofrontal cortex (LO) or agranular insular cor- adapting generalization gradients to changes in odor-
tex (AI). The C57BL/6 mice were perfused seven days after associated reward levels. Using computational modeling, we
CTb injection. The CTb iontophoretic injection created very are outlining a common framework for OB processing and
confined injection sites of a diameter of 150 to 200 µm. Our neuromodulation to understand and explain how OB-based
preliminary data reveal that for mice injected in the LO, CTb circuitry can instantiate appropriate topologies of learning
positive cells were found in endopiriform nucleus and the in response to experience. Acknowledgements: Supported by
ventral-medial area of layer II/III of the anterior piriform NIDCD grant DC009948.
#P93 POSTER SESSION II: OLFACTION Cornell University Department of Neurobiology and
DEVELOPMENT; TASTE CNS; NEUROIMAGING; Behavior Ithaca, NY, USA
OLFACTION CNS The olfactory bulb (OB) and piriform cortex (PC) receive
GABAergic gating of olfactory-motor transmission dense cholinergic projections from the diagonal band of
Broca in the basal forebrain. Cholinergic modulation within
Gheylen Daghfous1,2, Elias Atallah2, Jean-Luc Létourneau1, the PC has long been proposed to serve an important function
François Auclair2, Dominique Derjean1, Barbara S Zielinski3, in olfactory learning and memory. We here investigate
Réjean Dubuc1,2 how olfactory discrimination learning is regulated by
1
Groupe de Recherche en Activité Physique Adaptée cholinergic modulation of the OB inputs to the PC. Using
(GRAPA), Département de kinanthropologie, Université pharmacological manipulation of the OB, we examined the
du Québec à Montréal Montreal, QC, Canada, 2Groupe role of bulbar cholinergic signaling in rats’ performance on
de Recherche sur le Système Nerveux Central (GRSNC), a two-alternative choice odor discrimination task. Results
Département de physiologie, Université de Montréal show that blocking bulbar cholinergic signaling significantly
Montreal, QC, Canada, 3Department of Biological Sciences, slows learning, although the relative contribution of
University of Windsor Windsor, ON, Canada muscarinic (MAChRs) and nicotinic receptors (NAChRs)
depends on task difficulty. Specifically, blocking MAChRs
Olfactory stimuli induce and modulate locomotor activity
(38 mM scopolamine) impaired learning for nearly all odor
during vital behaviors such as homing, predator avoidance,
sets tested (n=13), whereas blocking NAChRs (19 mM
reproduction, foraging and feeding. The neural substrate
MLA) only affected learning when the task was made
underlying olfactory-motor behaviors was recently uncov-
difficult by using perceptually similar odors. This pattern
ered in sea lampreys (Petromyzon marinus) [Derjean et al.
of behavioral effects is consistent with predictions from a
2010 PLoS Biol 8(12): e1000567]. It consists of a specific neu-
recently developed model of cholinergic modulation in the
ral pathway, extending from the medial part of the olfactory
OB and PC (de Almeida et al., 2012). The model suggests
bulb (OB) to the mesencephalic locomotor region (MLR),
that MAChRs and NAChRs serve complementary roles
with a single relay in the posterior tuberculum (PT). In all
in regulating OB output and cortical learning. Namely,
vertebrates, the MLR acts as a motor command center that
NAChRs determine the output rate within each OB
controls locomotion via a descending projection to reticu-
channel and therefore regulate the overlap between learned
lospinal neurons (RS). This oligosynaptic pathway permits
representations in the cortical network. On the other hand,
movements to be rapidly generated in response to olfac-
MAChRs control the timing of spikes across OB output
tory stimuli, and thus functions as a pathway dedicated to
channels and, as a consequence, regulate the strength of
action. The modulatory mechanisms that act on this path-
odor representations in the cortical network. Together,
way and that are responsible for affecting the variability of
these results suggest that MAChRs in the OB serve a
the lamprey’s behavioral responses to olfactory inputs are
general role in regulating learning, whereas NAChRs are
still unknown. We addressed this question by using anatomi-
only critical when there is substantial overlap in the sensory
cal (tracers and immunohistochemistry) and physiological
inputs. Acknowledgements: NIH R01 DC009948 (CL)
(intracellular recordings) techniques. Retrograde axonal trac-
NIH F32 DC011974 (SD) L’Oreal Fellowship for Women
ing from the PT combined with GABA immunofluorescence
in Science (SD)
showed dense GABAergic innervation of the medial OB, a
central component of the olfactory-locomotor pathway,
suggesting a role for GABA in the modulation of this path- #P95 POSTER SESSION II: OLFACTION
way. Physiological experiments showed that injections of the DEVELOPMENT; TASTE CNS; NEUROIMAGING;
GABA antagonist gabazine (0.1 – 1 mM) into the medial OB OLFACTION CNS
considerably amplify or unmask the responses of RS neurons
to olfactory inputs. Taken together, our results suggest that Retronasal odor intensity coding in the dorsal olfactory
GABAergic innervation of the OB acts as a gatekeeper for bulb of rats
sensory inputs to motor control centers. Acknowledgements: Shree Hari Gautam1,2, Michelle R Rebello1, Justus V
Great Lakes Fishery Commission CIHR NSERC FRSQ Verhagen1
John B Pierce Lab New Haven, CT, USA, 2University of
1
Arkansas Fayetteville, AR, USA
OLFACTION CNS In nature food contains many volatile chemicals with a wide
range of concentrations. The volatiles, when released in the
Distinct roles of bulbar muscarinic and nicotinic receptors in mouth while eating, travel to the nasal cavity via the naso-
olfactory discrimination learning
pharynx evoking a retronasal smell which contributes to food
Sasha Devore, Licurgo de Almeida, Christiane Linster flavor. The olfactory system is responsible for encoding not
only the quality but also the concentration of the volatiles dependent on both intra and extracellular calcium, which
present in food. It is believed that each odor is represented maintains rhythmicity. The oscillatory discharge patterns
by a unique glomerular activation pattern in the olfactory observed in AOB mitral cells could play an important role in
bulb. However, whether and how retronasal odor concen- the signal coding and/or hormonal homeostasis controlled by
tration is encoded by the spatiotemporal activity pattern of mitral cells target regions in higher brain centers.
olfactory glomeruli, without confounding the quality of a
different odorant, remains unknown. In this study we opti-
cally imaged the retronasal odor-induced calcium responses #P97 POSTER SESSION II: OLFACTION
of olfactory receptor neurons in the dorsal olfactory bulb DEVELOPMENT; TASTE CNS; NEUROIMAGING;
in double-tracheotomized rats. We found reliable concentra- OLFACTION CNS
tion-response curves that differed between odors. MDS of Lateralized differences in olfactory bulb volume relate to
the spatial OB patterns suggest that ambiguity among select lateralized differences in olfactory function
stimuli may occur. Further, the relation between dynam-
ics and concentration differed remarkably among retrona- Thomas Hummel1, Antje Haehner1, Cornelia B Hummel1,
sal odorants. Understanding of coding for retronasal odor Ilona Croy2, Emilia Iannilli1
intensity has potentially important implications in the feed- Smell & Taste Clinic, Dept. of ORL, Technical Univ of
1
ing behavior and flavor neuroscience. Dresden Dresden, Germany, 2Dept. of Psychosomatic
Medicine, Technical Univ of Dresden Dreden, Germany
The present study aimed to investigate whether side
#P96 POSTER SESSION II: OLFACTION differences in olfactory bulb (OB) volume correlate to
DEVELOPMENT; TASTE CNS; NEUROIMAGING; respective differences in olfactory function. In a total of
OLFACTION CNS 164 healthy volunteers volumetric measures of the OBs
Intrinsic oscillatory discharge patterns in mitral cells of the were performed plus lateralized measurements of odor
mouse accessory olfactory bulb thresholds and odor discrimination. Side differences were
defined as 10% difference between the left and right OB. In
Monika Gorin, Marc Spehr
39 cases volumes on the right side were larger than on the
Dept. of Chemosensation, Institute of Biology II, RWTH left side, whereas in 29 cases it was the other way around.
Aachen University Aachen, Germany Subjects with larger right-sided OB volumes were found
to be more sensitive to odorous stimulation of the right as
The accessory olfactory bulb (AOB) represents the first stage
compared to the left nostril in terms of odor thresholds and
of central information processing in the rodent accessory
odor detection; while correspondingly, higher sensitivity
olfactory system. In the vomeronasal organ, social chemosig-
of left nostrils was observed in individuals with larger OB
nals activate sensory neurons which form synaptic contacts
volumes on the left side. These data appear to suggest that
with mitral/tufted cells, the main excitatory projection neu-
OB volume is partly dependent on lateralized influences on
rons in AOB. Bypassing the thalamo-cortical axis, these neu-
the olfactory system, reflecting its lateralized organization.
rons project directly to higher brain regions such as amygdala
Acknowledgements: Supported by a grant from the “Roland
and hypothalamus. Despite their physiological significance,
Ernst Stiftung” to TH.
the intrinsic properties of mitral cells and their role in social
information coding and signal integration in the AOB are not
fully understood. Here, we investigate the biophysical prop- #P98 POSTER SESSION II: OLFACTION
erties of AOB mitral cells using both voltage- and current- DEVELOPMENT; TASTE CNS; NEUROIMAGING;
clamp whole cell recordings from optically identified neurons OLFACTION CNS
in acute mouse AOB tissue slices. We identify a population
of mitral cells that display slow oscillatory discharge pat- Olfactory Sensory Neuron Physiology and Exposure-
terns which persist after pharmacological inhibition of syn- Induced Plasticity Are Altered in Adult Olfactory Marker
aptic transmission (AP5, NBQX and gabazine), revealing Protein Knockout Mice
the network-independent origin of this bursting behavior. Marley D. Kass, Andrew H. Moberly, John P. McGann
The underlying subthreshold membrane potential fluctua-
Rutgers University/Psychology Department New Brunswick,
tions with alternating up/down states display a high degree
NJ, USA
of periodicity. Using electrophysiological and pharmaco-
logical approaches, we analyze the basic ionic mechanisms Olfactory marker protein (OMP) is highly and selectively
underlying mitral cell oscillatory discharge. Our data dem- expressed in olfactory sensory neurons (OSNs) across spe-
onstrate a complex interplay of multiple voltage-gated ionic cies, but its function remains elusive. Previous in vitro studies
conductances, such as TTX-sensitive sodium channels and of MOR23-expressing OSNs suggested that OMP accel-
TEA-sensitive potassium channels as well as conductances erates the OSNs’ response to odorants and may modulate
the odorant- selectivity of OSNs (Lee et al. 2011 J Neurosci analyses of major neuropile volumes. We counted the number
31:2974–82). Here we performed in vivo optical imaging of functional units (glomeruli) within the antennal lobe, the
in adult mice expressing the fluorescent exocytosis indica- first olfactory neuropile in insects, and quantified synaptic
tor synaptopHluorin from the OMP locus. We compared structures in higher-order sensory integration centers
the spatiotemporal patterns of odor-evoked transmitter (mushroom bodies). The investigated Osmia species showed
release from OSNs in mice that were heterozygous for OMP significant differences in selected neuropile volumes and also a
(OMP−/+) or OMP-null (OMP−/−) and found that these large interspecies variance in glomerular numbers, correlated
patterns developed on a slower timescale in OMP−/− mice to floral preference. The strictly oligolectic species Osmia
but eventually reached the same magnitude as in OMP−/+ adunca showed the smallest number of glomeruli, whereas
mice. In OMP−/− mice, OSNs innervating a given glomeru- all polylectic species showed larger glomerular numbers.
lus also responded to a broader range of odorants than in The mushroom bodies of polylectic and oligolectic species
OMP−/+ mice. These results extend the previous in vitro showed the same density of synaptic structures, but expressed
findings in MOR23-expressing OSNs to other OSN popu- significant volume differences in the subregions that process
lations in vivo. We next evaluated the effects of a 7 day olfactory information. Chemical analyzes of host-plant odors
odor exposure paradigm on these spatiotemporal patterns and behavioral tests will be next steps to understand the large
in adult OMP−/+ and OMP−/− mice. We found that in impact of floral preference on the complexity of the olfactory
OMP−/+ mice, odorant exposure reduced the number of system in bees. Acknowledgements: DFG KE-1701 1/1
glomeruli receiving OSN input evoked by the exposure odor-
ant and the magnitude of those inputs but had no effect on
the response to unrelated odorants. In contrast, in OMP−/− #P100 POSTER SESSION II: OLFACTION
mice this experience-dependent suppression was observed DEVELOPMENT; TASTE CNS; NEUROIMAGING;
in the responses to all test odorants, not just the exposure OLFACTION CNS
odorant. These results suggest that OMP not only conveys
odor-selectivity on OSNs but also plays a role in restricting Temporal-spatial transformation in the piriform cortex
experience-dependent plasticity to specific OSN popula- Alex Koulakov1, Honi Sanders2, Brian Kolterman1, Dima
tions. Acknowledgements: This work was supported by the Rinberg3, John Lisman1
National Institute on Deafness and Other Communication
Cold Spring Harbor Laboratory Cold Spring Harbor, NY,
1
Disorders (R00 DC009442 to JPM).
USA, 2Brandeis University Waltham, MA, USA, 3New York
University New York, NY, USA
#P99 POSTER SESSION II: OLFACTION Mitral cells of the olfactory bulb respond to stimuli with brief
DEVELOPMENT; TASTE CNS; NEUROIMAGING; and temporally precise transient changes in the firing rate
OLFACTION CNS (sharp events) that tile the inhalation cycle. This suggests that
information about odorants can be encoded by the temporal
Floral Preference is reflected in the Neuroanatomy of the sequence of these events. Here we propose a class of compu-
Olfactory System in Mason Bees (Osmia) tational models for the olfactory cortex that can detect such
Christina Kelber1, Thomas Schmitt1, Wolfgang Roessler2 sequences and convert them into a spatial pattern that can
be recognized by standard attractor networks. We propose
1
Department of Biology, Ecological Networks TU Darmstadt,
that the olfactory cortex contains groups of cells that can be
Germany, 2Department of Behavioral Physiology and
sequentially activated by inputs from mitral cells synchronized
Sociobiology, Biozentrum University of Wuerzburg,
at different phases of the respiratory cycle. Neurons in each
Germany
group can be persistently activated by virtue of, for example,
Locating food sources is important for all insects, and in many an intrinsic bistability mechanism. The pattern of activation
species the olfactory system is crucial for finding suitable of neurons in each group carries a snapshot of coincidences in
sources. Two main strategies can be found: either to be able mitral cell sharp events at a particular phase of the breathing
to use many different food sources (generalists) or to be cycle. Due to long-range intracortical connectivity, the activa-
specialized on a single or only few food sources (specialists). tion of one group “enables” bistability in another group which
In bees, both generalist and specialized species can be found can then form a snapshot of mitral cell activity at a later phase
that collect pollen and nectar either from many plant families of the respiratory cycle. In this way, persistent activation of
(polylectic) or from only few species (oligolectic). In some groups of neuron occurs sequentially, each in turn representing
cases, both types of food preferences can be found within the olfactory bulb activity at a certain phase of the sequence.
the same genus, like in the mason bee genus Osmia. Here We further show that sharp events in mitral cell responses
we investigate how the floral preference is reflected in the occur at a preferred phase of gamma cycles (measured in the
neuroanatomy of the olfactory system. We employed confocal field potential). Given that there are only a few gamma cycles
microscopy scanning and 3D-reconstruction for quantitative within a sniff, the number of groups needed to define gamma
cycle specific snapshots of an odorant is not large. Recognition University of Arizona/Department of Neuroscience Tucson,
may occur when the spatial pattern becomes sufficient to dis- AZ, USA
tinguish among the potential odorants. The male moths of Manduca sexta are more attracted to a
mimic of its natural female sex pheromones, composing of
#P101 POSTER SESSION II: OLFACTION only two essential components in a ratio that is found in its
natural pheromones. Deviation from this ratio causes reduced
behavior. The projection neurons innervating the pheromone
OLFACTION CNS
responsive region of the male antennal lobe produce maxi-
Unique Cholinergic Interneuron Populations in the Mouse mal synchronized spiking activity in response to blends con-
Accessory Olfactory Bulb: Neurochemical Expression and sisting of the two components centering around the natural
Fiber Density ratio, leading to a hypothesis that blend ratios are encoded in
Kurt Krosnowski, Sarah Ashby, Weihong Lin neuronal synchrony. To test this hypothesis, we investigated
the physiological and morphological features of down-stream
University of Maryland Baltimore County Baltimore, protocerebral neurons that were challenged with stimulation
MD, USA of single pheromone components and their blend of differ-
The accessory olfactory bulb (AOB) is a primary central ent ratios. We found a small proportion of protocerebral neu-
processing site of sensory information detected via the rons showing stronger responses to the blend of natural ratio
vomeronasal organ. The AOB contains diverse populations whereas many other neurons did not distinguish these blends
of intrinsic interneurons. We detected a largely unidentified at all. In a multi-dimensional analysis, we also found the
choline acetyltransferase-expressing (ChAT) cholinergic population response mapped onto the second principle axis
interneuron population using ChAT(BAC)-eGFP mice. Here displayed most distinction among the two pheromone com-
we classified their neurochemical expression and distribution ponents and their blend, and the distinction occurred prior
throughout the AOB. We then determined if this cholinergic to the peak population response - a result consistent with
interneuron population differs from other known populations an earlier observation where neural synchrony in the anten-
of interneurons in the AOB and main olfactory bulb (MOB). nal lobe tends to maximize before the firing rate reaches its
Similar to the MOB (Krosnowski et al 2012), we found that peak. Moreover, the response patterns of protocerebral neu-
all cholinergic interneurons are neither dopaminergic nor rons are very diverse, indicating the complexity of internal
GABAergic. While most ChAT expressing cells in the external representation of odor stimuli at the level of protocerebrum.
plexiform layer (EPL) of the AOB are not glutamatergic, we Acknowledgements: This work was supported by NSF grant
found some coexpression between ChAT-GFP and GluR2/3, a DMS-1200004 to HL, NIH grant R01-DC-02751 to JGH
glutamatergic marker, in contrast with results obtained from the
MOB. Also, unlike the cholinergic interneuron population in
the MOB, the majority of cholinergic interneurons in the AOB
#P103 POSTER SESSION II:OLFACTION
do not express a calcium binding protein, calbindin-D28K. DEVELOPMENT; TASTE CNS; NEUROIMAGING;
Further, clear differences can be seen between cholinergic OLFACTION CNS
nerve fibers in the internal plexiform layer (IPL) of the MOB Comparison of changes in odor-induced firing of mitral cells
and the AOB. Unlike in the MOB, where the highest density of and oscillations in the local field potentials in mice learning
cholinergic nerve fibers was found in the IPL, in the AOB, the to discriminate odors
IPL contains the fewest visible fibers. Instead, the majority of
Anan Li, Diego Restrepo
cholinergic fibers in the AOB are found in the EPL. Thus, our
data supports the idea that the intrinsic cholinergic interneuron Department of Cell and Developmental Biology, Rocky
populations in the AOB are distinct from previously identified Mountain Taste and Smell Center and Neuroscience
interneuron populations in both the MOB and AOB and this Program Aurora, CO, USA
suggests that they play a unique role in signal processing in the Odor induced mitral cell firing and changes in local field
accessory olfactory system. Acknowledgements: NIH/NIDCD potentials (LFPs) are modified as an animal learns to dis-
009269, 012831 and ARRA administrative supplement to WL criminate between odors. In previous work we reported
that as the animal learns to discriminate between odors in
go-no go odor discrimination tasks synchronized unit fir-
#P102 POSTER SESSION II:OLFACTION ing of mitral cells develop divergent responses to rewarded
DEVELOPMENT; TASTE CNS; NEUROIMAGING; and unrewarded odors, and convey important informa-
OLFACTION CNS tion on odor quality in addition to odor identity in awake
behaving mice (Doucette et. al. Neuron 69, 1176–1187,
Blend Processing by Protocerebral Neurons of Manduca sexta
2011). LFPs reflect integrated signals from cell ensembles
Hong Lei, Hong-Yan Chiu, John Hildebrand also show divergent responses. However, how mitral cell
firing and local LFPs are related and more importantly #P105 POSTER SESSION II: OLFACTION
how these are related on a trial-by-trial basis when the DEVELOPMENT; TASTE CNS; NEUROIMAGING;
animal makes mistakes remains to be elucidated. Here OLFACTION CNS
our preliminary data indicate that unit firing and beta
oscillations of LFPs (10–35 Hz) show related changes Influences of lateral amygdala activation on piriform
during the learning process of the go-no go task: at the cortical odor processing
beginning of the task, there is no or very weak divergent Benjamin Sadrian1,2, Donald Wilson1,2
odor responses for both signals, while obvious and strong 1
NYU School of Medicine New York, NY, USA, 2Nathan Kline
divergent responses are found as the mice learn to dis-
Institute Orageburg, NY, USA
criminate the odor pairs. Acknowledgements: DC00566
and DC04657 Olfactory sensory processing in the piriform cortex requires
the synergy of odorant ligand input, local inhibitory feed-
#P104 POSTER SESSION II: OLFACTION back loops, and interregional modulation, in order to synthe-
DEVELOPMENT; TASTE CNS; NEUROIMAGING; size emotionally relevant and contextually significant odor
OLFACTION CNS percepts. Reciprocal connectivity between the piriform cor-
tex and higher processing regions, such as the lateral entorhi-
Identification of Microglia in the Peripheral Deafferentation nal cortex and amygdala, provide currently understudied
Response of the Adult Zebrafish Olfactory Bulb routes through which odor processing in the piriform may
Amanda K McKenna, Christine A Byrd-Jacobs be regulated. We have employed optogenetic techniques to
investigate how activation of lateral amygdala (LA) during
Western Michigan University/Biological Sciences Kalamazoo, odor presentation affects piriform cortical odor processing.
MI, USA We have performed single unit recordings of both sponta-
Our lab has been examining the potential role of microglia neous and odor-evoked activity in the anterior piriform
in the deafferentation response of the zebrafish olfactory cortex, and summarized a range of LA-influenced changes
bulb. We previously used phagocytosis-dependent labeling in piriform activity. We have also begun using a fear con-
with DiA to illustrate the putative microglial response ditioning model to investigate the influences of emotional
following olfactory organ ablation. DiA-labeled puncta in significance on odor processing in the piriform. We aim to
the deafferented olfactory bulb increased dramatically in describe how such contextual changes affect the precision of
number and then diminished over the course of a week. both cortical odor processing and behavioral odor percep-
The labeling pattern corresponded directly to areas of the tion. Acknowledgements: T32-MH067763 from the NIMH
bulb with damaged axons. In that study, we were unable to B.A.S. and R01-DC003906 from the NIDCD to D.A.W.
to identify the as microglia. The current study seeks to
confirm both the presence and active role of microglia in #P106 POSTER SESSION II: OLFACTION
the deafferented zebrafish olfactory bulb using an antibody DEVELOPMENT; TASTE CNS; NEUROIMAGING;
to zebrafish microglia (anti-4C4). Zebrafish were treated OLFACTION CNS
either with cautery ablation or Triton X-100 application
to the olfactory organ to cause either permanent or Wild Scents: comparing the olfactory anatomy of caged and
temporary deafferentation of the bulb. We hypothesized wild mice
that the pattern of anti-4C4 labeling would mimic the Ernesto Salcedo, Kyle Hanson, Taylor Jonas, Lois Low
pattern seen with DiA. We found that the olfactory bulb
Diego Restrepo University of Colorado School of Medicine
had an obvious increase in 4C4-positive microglia 1 day
Aurora, CO, USA
following both permanent and temporary treatments.
These 4C4-positive profiles had primarily amoeboid We have previously detailed the subtle neuroanatomical
morphology; they were found throughout the bulb layers changes we found in the glomeruli of olfactory bulbs from
but were concentrated around the degenerating axons. genetically identical mice reared in cages with different
Over the next several days, the 4C4-positive microglia levels of ventilation (Oliva and Salcedo et al, 2010). In these
appeared to decrease in number; they also changed to mice, we were able to correlate these glomerular changes
mostly ramified morphologies. This pattern overlaps with marked increases in aggressive behavior towards
with the DiA results but also appears to show additional invader mice, highlighting the exquisite sensitivity a mouse’s
microglia not actively phagocytizing axonal debris. Thus, olfactory neuroanatomy has to its environment. In order to
there is a profound microglial response immediately after examine the broader effects that environment may have on
both permanent and temporary deafferentation in the the formation of the olfactory system, we have trapped wild
adult zebrafish olfactory bulb that sharply declines over house mice from the Denver environs and have rigorously
the next several days. Acknowledgements: Supported by characterized the neuroanatomy of their main olfactory
NIH- NIDCD #011137 (CBJ) bulbs (MOB) using MATLAB mapping software developed
in-house and immunohistochemical techniques. On gross epithelium of mice was assessed in frozen sections under
examination, the MOBs from the wild mice do not appear a fluoroscopic microscope. Results: The ratio of uptake
to be significantly different from their caged brethren. Nor of nasally administered 125I-IGF-I in the cerebrum to
did we find any significant immunostaining differences in uptake in the blood of the model group was significantly
OMP of GAP43 labeling of the MOB. Although somewhat decreased compared to the control group. The accumula-
smaller, the wild olfactory bulbs had an estimated number of tion of nasally administered neuronal tracer in the nasal
glomeruli (using Meisami’s Correction) that does not differ epithelium of mice was significantly prevented by the
significantly from the estimated number of glomeruli found in resection of the olfactory bulb. Conclusions: Olfactory
the MOBs of their caged counterparts. Curiously, we do find bulb resection results in the reduced delivery of nasally
a dramatic difference in the distribution of olfactory sensory administered IGF-I to the brain due to the disconnection
innervations across the surface of the MOB: caged mice of the olfactory nerve between the nasal epithelium and
tended to have larger glomeruli that occupied a significantly olfactory bulb in vivo. Acknowledgements: Grant-in-Aid
larger portion of the glomerular layer then did the wild for Scientific Research from the Ministry of Education,
mice. This distribution was particularly pronounced in the Science and Culture of Japan (C21592174 to H.S.) and
ventro-medial portion of the bulb around the AOB. These Assist Kaken from Kanazawa Medical University (J.Y.)
results provide further evidence that olfactory environment
plays a role in fine-tuning the formation and maintenance
of glomeruli in the main olfactory bulb. Acknowledgements: #P108 POSTER SESSION II: OLFACTION
NIDCD DEVELOPMENT; TASTE CNS; NEUROIMAGING;
OLFACTION CNS
Dynamics of reward-mediated neural plasticity in honey
bee antennal lobe glomerulus
OLFACTION CNS Adrian Smith1,4, Irina Sinakevitch1, Ramon Huerta2, Maxim
Bazhenov3, Brian H Smith1
Assessment of nasally administered insulin-like growth
factor-I accumulation in the cerebrum of mice with resected
1
Arizona State University, School of Life Science, Tempe,
olfactory bulb AZ, USA, 2BioCircuits Institute, University of California San
Diego, La Jolla, CA, USA, 3Department of Cell Biology and
Hideaki Shiga1,2, Mikiya Nagaoka2, Kohshin Washiyama2,
Neuroscience, University of California, Riverside, CA, USA,
Junpei Yamamoto1, Ryohei Amano2, Takaki Miwa1 4
Arizona State University, Mathematical, Computational and
1
Otorhinolaryngology, Kanazawa Medical University Modeling Sciences Center, Applied Mathematics for the Life
Ishikawa, Japan, 2Quantum Medical Technology, Kanazawa and Social Sciences Tempe, AZ, USA
University Ishikawa, Japan
Recent studies of neural plasticity in the honey bee antennal
Objectives: To show the role of the olfactory bulb in the lobe (AL) show the response dynamics of uniglomerular
delivery of nasally administered insulin-like growth fac- projection neurons (uPNs) to an odor change after
tor-I (IGF-I) to the brain in vivo. Nasal administration association of that odor with sucrose reinforcement. The
of IGF-I has been shown to enable drug delivery to the octopamine (OA), released by the ventral unpaired median
brain beyond the blood brain barrier in vivo. IGF-I is neuron (VUM), is necessary for these neural plasticity
associated with the development and growth of the central changes. Using anti-OA staining, we found that the
nerve. Methods: The ratio of uptake of nasally adminis- varicosity-like distributions of VUM branch mostly in the
tered 125I-IGF-I in the cerebrum to uptake in the blood of cortex of the glomerulus, where it potentially modulates
male ICR mice with resected left olfactory bulb (8 weeks olfactory receptor neurons (ORNs), local interneurons
of age, the model mice) was compared to that of the sham- (LNs) and uPNs. We develop a biophysical model of the
operated male ICR mice (8 weeks of age, the control mice). AL circuit to investigate modulatory mechanisms that can
We exposed and resected the left olfactory bulb, cutting explain existing data on the dynamical changes of uPNs
the frontal bones of model mice, and just exposed the left during associative learning, and lead to insights for new
olfactory bulb in control mice under anesthesia. 125I-IGF-I experiments. First, we hypothesize that OA release from
(human, recombinant) saline solution was obtained from VUM varicosities would be dependent on correlated firing
PerkinElmer Japan (Yokohama, Japan), and 10µl was between ORNs and VUM during associative learning.
instilled into the left nostril of each mouse with a micro- This mechanism implies simultaneous cholinergic and
injection pipette under anesthesia. The radioactivity of octopaminergic transmission to PNs. Second, OA release
the samples was measured with gamma spectrometry. from VUM acts on AmOA1 receptors expressed in LNs.
The accumulation of the nasally administered neuronal AmOA1 activation increases the excitability of LNs,
tracer (fluoro-ruby; dextran tetramethylrhodamine) in the leading to increased inhibition of PNs. Third, release of
OA leads to direct activation of beta-adrenergic-like OA #P110 POSTER SESSION II: OLFACTION

receptors. This increases the levels of cAMP, triggering DEVELOPMENT; TASTE CNS; NEUROIMAGING;
PKA-dependent translation and upregulation of alpha7 OLFACTION CNS
nicotinic acetylcholine receptor (nAChR) subtypes.
nAChR-dependent Ca2+ influx triggers transcription Expression and Activity of Glucagon-like Peptide-1 in the
factors that upregulate transient Shal-type K+ channels, Mouse Olfactory Bulb
preventing excessive membrane depolarization. Based on Nicolas Thiebaud1, Ida Llewellyn-Smith2, Fiona Gribble3,
these hypotheses, we propose the model to characterize Frank Reimann3, Stefan Trapp4, Debra A. Fadool5
dynamics of the uPN as a function of the relative 1
The Florida State University, Department of Biological
expression of Shal-type K+ channels and OA release.
Science Tallahassee, FL, USA, 2Flinders University,
Acknowledgements: NIH-NCRR RR014166 and NIH
Centre for Neuroscience Bedford Park, Australia,
grant R01 DC011422 3
Addenbrooke’s Hospital, Cambridge Institute for
Medical Research Cambridge, United Kingdom, 4Imperial
College London, Department of Surgery and Cancer
#P109 POSTER SESSION II: OLFACTION London, United Kingdom, 5The Florida State University,
DEVELOPMENT; TASTE CNS; NEUROIMAGING; Program in Neuroscience and Molecular Biophysics
OLFACTION CNS Tallahassee, FL, USA
In vivo imaging of odor-evoked responses in the mouse A number of peptides and hormones that are known to
olfactory bulb using the FP voltage sensor ArcLight and the regulate energy metabolism or feeding behavior have been
calcium sensor GCaMP3 identified in the olfactory system. These hormones are
thought to modulate olfactory perception and function
Douglas A Storace1, Lawrence B Cohen1, 2, Uhna Sung2 to suppress or promote appetite. Glucagon-like peptide-1
(GLP-1) is an incretin peptide that also suppresses food
1
Yale University / Department of Cellular and Molecular
intake, and in situ hybridization in rat has suggested that
Physiology New Haven, CT, USA, 2Korea Institute of Science
both GLP-1 and its receptor are present in the olfactory
and Technology / Center for Functional Connectomics Seoul,
bulb (OB). Furthermore, GLP-1 modulation of taste
South Korea
sensitivity has been reported for taste buds, prompting us
Optogenetic reporters of membrane potential allow for to speculate a similar role in the olfactory system. Using
recording of genetically distinct populations of neurons, transgenic mice expressing YFP under the preproglucagon
although their usefulness to date has been limited by poor (PPG) promoter, we confirm that a population of YFP-
in vivo expression, small signal sizes and slow kinetics. The immunoreactive neurons exists in the granule cell layer
fluorescent protein (FP) voltage sensor ArcLight exhib- (GCL) of the OB, indicative of GLP-1 producing cells.
its a change in fluorescence to a 100 mV depolarization The labeled neurons had a typical granule cell morphology
five times larger than previously reported probes in HEK with a single axon projecting into the adjacent mitral cell
293 cells. However, recordings of ArcLight in mammalian layer (MCL). We observed strong immunoreactivity for
neurons have been limited to cultured neurons. The goal the GLP-1 receptor in the MCL and in sparse cells in the
of the present study was to examine ArcLight responses in GCL in olfactory marker protein (OMP-GFP) transgenic
the olfactory bulb in an in vivo preparation, and compare mice. Binding of biotinylated GLP-1 to the GLP-1 receptor
them to those of the genetically encoded calcium indicator was visualized within the same regions. We confirmed the
GCaMP3. AAV-1 viral transduction was used to express presence of functional GLP-1 receptors on mitral cells
ArcLight and GCaMP3 in the mouse olfactory bulb. Odors (MCs) using whole-cell patch-clamp recordings in acute OB
were presented at different stimulus duration and concentra- slices. Bath perfusion of 1 µM GLP-1 or its stable analogue,
tions, and the resulting patterns of activation were imaged. exendin-4, resulted in a significant increase in the evoked
Odor-specific patterns of activation were obtained from both action potential frequency (370% for GLP-1 and 170% for
ArcLight and GCaMP3, although only ArcLight had suf- exendin-4) and a concomitant decrease in the interburst
ficiently fast temporal kinetics to clearly detect population interval in about 60% of the sampled MCs. These results
activity elicited by individual sniffs to an odor. The results show that GLP-1 is synthesized locally in the OB and
indicate that ArcLight can be used as a reliable detector directly affects the firing properties of MCs, suggesting a
of odor-evoked population signals in the mouse olfactory potential paracrine modulation of olfactory output with
bulb. Acknowledgements: Supported by US NIH Grants satiety. Acknowledgements: This work was supported by
DC005259 and NS057631, Grant WCI 2009-003 from the R01 DC003387 from the NIH/NIDCD, a Creative Research
National Research Foundation of Korea, and an James Council (CRC) award from FSU, a Project Grant 1025031
Hudson Brown – Alexander Brown Coxe Fellowship from from NHMRC Australia, and MR/J013293/1 from the
Yale University. MRC United Kingdom.
#P111 POSTER SESSION II: OLFACTION Diabetes and Nutrition Baltimore, MD, USA, 4University of
DEVELOPMENT; TASTE CNS; NEUROIMAGING; Maryland School of Medicine Baltimore, MD, USA
OLFACTION CNS The main olfactory system has distinct subsystems that dif-
Enhanced Survival of Newly Formed Cells Contributes to fer in the chemosensory stimuli to which they respond and
Restoration of Olfactory Bulb Volume Following Reversible the connections they make to the brain. In contrast to the
Deafferentation in Adult Zebrafish canonical glomeruli of the main olfactory bulb (MOB),
individual necklace glomeruli (NGs) receive heterogene-
Darcy M Trimpe, Christine A Byrd-Jacobs ous olfactory sensory neuron (OSN) innervation [including
Western Michigan University/Biological Sciences Kalamazoo, semiochemical-sensitive OSNs expressing guanylyl cyclase D
MI, USA (GC-D)] and display extensive intrabulbar connections. This
organization suggests that NGs integrate multiple olfactory
Our lab has shown that chronic partial deafferentation, signals. To better understand the functional circuitry associ-
achieved through unilateral chemical ablation of the olfac- ated with NGs, we examine the transsynaptic spread of neu-
tory epithelium with Triton X-100, results in a decrease in ronal activity using intrinsic flavoprotein fluorescence (FF)
olfactory bulb volume, while cessation of treatment allows imaging. FF imaging measures the changes in endogenous
bulb volume to recover. We hypothesized that alterations in fluorescence produced by mitochondrial flavoproteins that
cell genesis and/ or survival would be involved in restoration accompany increased metabolic demand. The FF signals
of olfactory bulb size. Bromodeoxyuridine (BrdU) adminis- correspond to neuronal activity and can be followed across
tration was used to examine newly formed cells in the brain synapses, thus facilitating the mapping of functional circuits.
of adult zebrafish, with short-term survival allowing investi- Studies in horizontal MOB slices from 3–5 w.o. mice exhibit
gation of patterns of cell proliferation and long-term survival robust FF signal spread from the glomerular layer (GL) to
allowing examination of cell survival and fate. We first com- the external plexiform layer (EPL) following electrical stimu-
pared two methods of BrdU administration: immersion of lation (1–4 s, 10–50 Hz, 10–100 µA) of individual canonical
fish in the drug versus intraperitoneal injection. While both and necklace glomeruli. Bath application of 10 mM gabazine
methods revealed similar numbers of newly formed cells, increased lateral stimulus-dependent FF signal spread in
injection of the drug resulted in loss of fewer fish during treat- the GL/ EPL, and resulted in a 2-fold increase in FF signal
ment. Next, we examined potential alterations in cell genesis amplitude in the EPL. Single NG stimulation (from mice
and/or cell survival resulting from reversible partial deafferen- expressing green fluorescent protein under control of the
tation. Repeated detergent treatment followed by BrdU expo- gene encoding GC- D) results in reduced FF signal ampli-
sure showed no difference in the number of dividing cells in tude but prolonged FA signal duration compared to canoni-
the olfactory bulb, indicating that cell genesis is not affected. cal glomeruli. The use of FF imaging should help reveal
There was, however, an increase in newly formed cells that basic strategies of information processing in the MOB and
survived when the detergent treatment ceased, indicating that its subsystems. Acknowledgements: NIDCD (DC005633),
cell survival contributes to the restoration of bulb volume NIGMS (GM008181), NINDS (NS063391)
during the period of reinnervation. When fish were exposed
to BrdU before the repeated detergent treatment, there
appeared to be no effect on the number of newly formed cells. #P113 POSTER SESSION II: OLFACTION
Thus, enhanced cell survival, rather than cell genesis, appears DEVELOPMENT; TASTE CNS; NEUROIMAGING;
to be a contributing factor in the restoration of olfactory bulb OLFACTION CNS
volume following return of innervation in a reversible deaf-
ferentation model. Acknowledgements: Supported by NIH- DIFFERENTIAL MODIFICATIONS OF SYNAPTIC WEIGHTS
NIDCD #011137 (CBJ) DURING ODOR RULE LEARNING: DYNAMICS OF
INTERACTION BETWEEN THE PIRIFORM CORTEX WITH
LOWER AND HIGHER BRAIN AREAS
#P112 POSTER SESSION II: OLFACTION Yaniv Cohen1,2,3, Donald A. Wilson2,3, Edi Barkai1
1
Departments of Biology and Neurobiology, Faculty
OLFACTION CNS
of Natural Sciences, University of Haifa Haifa, Israel,
Characterizing Olfactory Bulb Circuitry using Intrinsic 2
Department of Child and Adolescent Psychiatry, New York
Flavoprotein Fluorescence Imaging University Langone School of Medicine New York, NY,
Cedric R Uytingco1,2,4, Adam C Puche1,2,4, Steven D USA, 3Emotional Brain Institute, Nathan Kline Institute for
Munger1,2,3,4 Psychiatric Research, Orangeburg New York, NY, USA
1
Depatment of Anaotmy and Neurobiology Baltimore, Learning of a particularly difficult olfactory-discrimi-
MD, USA, 2Program in Neuroscience Baltimore, MD, USA, nation (OD) task results in acquisition of rule-learning,
3
Department of Medicine, Division of Endocrinology, a process that requires prolonged and extensive training.
Previously, we demonstrated enhanced synaptic connectiv- in heptane) were 0.35, 0.07, 0.05 and 0 ppm (during 15,
ity between the piriform cortex (PC) and its ascending and 45, 60 and 30 min, respectively). During the 30 minutes
descending inputs from the olfactory bulb (OB) and orbit- of exposure subjects were exposed to only heptane at
ofrontal cortex (OFC) following OD rule learning. Here, the same concentration as in the other exposures (4.9
using recordings of evoked field post-synaptic potentials ppm). During exposure, eye irritation was rated on
in behaving animals, we examined the dynamics by which Borg’s CR-100 scale. Human exposure to acrolein at sub-
synaptic connectivity from the OB and OFC to the PC are threshold concentrations showed a cumulative effect on
modified during rule acquisition. We show profound differ- sensory irritation. During exposure to 0.35 ppm (but not
ences in the dynamics and strength of synaptic connectivity 0.07 and 0.05 ppm) acrolein evoked a significant increase
modulation between the ascending and descending inputs. in irritation compared to the control condition after about
During rule learning acquisition, the ascending synaptic 12 minutes of exposure. During exposure to 0.07 and
connectivity from the OB to the anterior and posterior PC 0.05 ppm only some of the subjects reported increased
is simultaneously enhanced. Notably, the daily OB electri- irritation after about 30 minutes of exposure. A large
cal stimulation used to examine the strength of synaptic variability in reported sensory irritation was seen between
inputs enhanced the rate of rule learning. In sharp con- individuals and this may be due to individual differences
trast, the synaptic input in the descending pathway from the in the ability to remove the electrophilic irritants from
OFC was significantly reduced during rule learning acqui- the cornea. Acknowledgements: The Swedish Research
sition. OFC stimulation had no effect on the rate at which Council FORMAS
the rule was acquired. Once rule learning was established,
the strength of synaptic connectivity in the two pathways
resumed its pre-training values. We suggest that acquisition #P115 POSTER SESSION III:TRIGEMINAL;
of olfactory rule learning requires a transient enhancement HUMAN OLFACTORY PSYCHOPHYSICS; TASTE
of ascending inputs to the PC, synchronized with a paral- PERIPHERY
lel decrease in the descending inputs. This combined short-
lived modulation is required to enable the PC network to Solitary Chemosensory Cells in Human Nasal Epithelium
reorganize in a manner that enables it to first acquire and Sarah E Cooper1, Marco Tizzano2, Vijay R Ramakrishnan1,
then maintain the rule. Henry P Barham1, Jameson K Mattingly1, Thomas E Finger1,2,
Sue C Kinnamon1,2
1
University of Colorado, Department of Otolaryngology
#P114 POSTER SESSION III: TRIGEMINAL; Aurora, CO, USA, 2University of Colorado, Department of
HUMAN OLFACTORY PSYCHOPHYSICS; TASTE Cell and Developmental Biology Aurora, CO, USA
PERIPHERY
Solitary chemosensory cells (SCCs) described in rodents
Human exposure to acrolein – time dependence on TRPA1 rely on the bitter taste transduction cascade to detect
agonists potential irritants within the airways. SCCs express all of
Anna-Sara Claeson, Nina Lind the taste GPCR signaling effectors including T2R bitter
taste receptors, α-gustducin, PLCβ2, and the transduction
Department od Psychology, Umeå university Umeå, Sweden
channel TRPM5. SCCs are innervated by the trigeminal
The objective of the study was to examine the time nerve and when stimulated evoke protective airway reflexes
dependence on sensory irritation potency of acrolein such as sneezing, apnea and local inflammation (Tizzano
(2-propenal) in humans. Concentrations at or below et al., 2010; AChemS 2012). We have begun investigating
earlier reported thresholds that initially are too low SCCs in human sinonasal epithelium and their clinical
to evoke sensory irritation in the eye but might do so implications. Previously we and others reported that cells
in exposures up to 60 minutes were used. Acrolein is a resembling SCCs occur in human biopsy material near the
known TRPA1 agonist present in cigarette smoke, smoke vestige of the vomeronasal organ (Braun et al., 2011) as
from fires, automobile exhaust and smog. The TRPA1 well as within the turbinates (Barham et al 2013; AChemS
channel is activated by electrophilic compounds that 2012). However, the exact distribution and abundance of
form covalent bonds with cysteine residues. Because of SCCs in humans is unknown. To map the distribution of
this mechanism of activation one can expect duration of SCCs, we obtained middle and inferior turbinate mucosa
exposure to be of importance in evoking sensory irritation. from human patients that were free of sinonasal disease, but
The exposures occurred in an exposure chamber and the were undergoing surgical procedures requiring removal of
subjects were breathing fresh air through a mask that this tissue. Whole mount tissue was stained with antibodies
covered the nose and mouth. All participants took part against TRPM5 and villin, which is expressed at the apex
in four exposure conditions, differing in duration and of microvillous, but not ciliated, epithelial cells. TRPM5-
concentration. The concentrations of acrolein (diluted immunoreactive cells were scattered heterogeneously in
the sinonasal tissue. The cells were most abundant on the #P117 POSTER SESSION III: TRIGEMINAL;
ridges of the turbinates and less abundant on the lateral HUMAN OLFACTORY PSYCHOPHYSICS; TASTE
margins. Many TRPM5 immunoreactive cells also labeled PERIPHERY
with the villin antibody, suggesting that TRPM5 is pre-
sent in microvillous but not ciliated cells of the epithelium. Cholinergic Regulation by ChAT and TrpM5 – Expressing
Studies are in progress to determine if disease state alters Microvillous Cells in Main Olfactory Epithelium and Solitary
the distribution or abundance of these cells and whether Chemosensory Cells in Nasal Cavity and Upper Airway
SCCs in humans are innervated by the trigeminal nerve, as Tatsuya Ogura, Steven A Szebenyi, Weihong Lin
in rodents. Acknowledgements: R01 DC009820 (TEF and Dept. of Biological Sciences, University of Maryland
SCK) P30 DC04657 (to D. Restrepo) Baltimore County Baltimore, MD, USA
Chemosensory detection of inhaled irritating, harmful sub-
#P116 POSTER SESSION III: TRIGEMINAL; stances in the nasal cavity and upper airway is essential for
vital organ protection. Previously, we identified a population
HUMAN OLFACTORY PSYCHOPHYSICS; TASTE
of micorvillous cells (MCs) in the main olfactory epithelium
PERIPHERY (MOE) that expresses both transient receptor potential chan-
Responses to change in oral temperature by neurons nel M5 (trpM5) and choline acetyltransferase (ChAT). Despite
in the mouse medullary dorsal horn and nucleus of the their distinct morphology and distribution, the MCs share
solitary tract properties, such as cell marker expression and chemosensibility
Yi Kang, Christian Lemon
with solitary chemosensory cells (SCCs) found in the nasal res-
piratory epithelium, vomeronasal duct and trachea (Lin et al.
St. Louis University St. Louis, MO, USA 2008, Ogura et al. 2010, 2011). Here, we provide comparative
Psychophysical data show the temperature of sapid studies of MCs and SCCs at different regions. Using single cell
solutions influences flavor. However, it is not entirely Ca2+ imaging, we compared intracellular Ca2+ response pro-
clear how central neural circuits for oral sensation encode file to chemical stimuli. We found that the order of respond-
temperature input. The trigeminal subnucleus caudalis ing cell percentage to various stimuli was similar among these
(Vc) is a brainstem somatic relay receiving temperature cells (high concentration odorants > denatonium > ATP >
signals from the oro-facial region and implicated for acetylcholine (ACh)). Because of the low percentage of ACh-
oral thermosensation. Additionally, the solitary tract sensitive MCs and SCCs, ACh released from these cells may
nucleus (NTS), the first central taste relay, also receives play a role in paracrine regulation to influence neighboring
thermal input from the mouth. Here we compared neural cells. Using Ca2+ imaging on intact epithelial preparations, we
responses of NTS and Vc to intraoral thermal stimulation found that ACh- induced increases in intracellular Ca2+ levels
in anesthetized mice to assess the contribution of activity in epithelial cells surrounding the MCs and SCCs were inhib-
in these structures to oral temperature responses in ited by muscarinic ACh antagonist atropine. Our results sug-
brain stem. Extracellular single-unit activities of NTS gest that MCs and SCCs share common physiological roles in
thermo-gustatory neurons and Vc thermo-somatic cells sensing chemical stimuli and may release ACh to influence sur-
were recorded after application of different temperature rounding non- sensory cells via paracrine mechanism. Because
stimuli to tongue, including cold (5 and 10 °C), cool/ MCs lack afferent innervation, this cholinergic paracrine
ambient (17 and 23 °C), and warm/ hot (30, 45, and regulation could be especially important for chemoreception-
48 °C). Temperature stimulation was achieved rapidly mediated regulation of MOE activities. Acknowledgements:
by oral flow of thermally varied water. Seventy- two Supported by research grants NIH/NIDCD 009269, 012831
neurons were obtained; 34 from Vc and 38 from NTS. and ARRA administrative supplement to WL
Analyses revealed significant differences between NTS
and Vc in responses to thermal stimuli [F(1, 70) = 5.80, #P118 POSTER SESSION III: TRIGEMINAL;
P<0.05], and a significant interaction between nucleus and HUMAN OLFACTORY PSYCHOPHYSICS; TASTE
temperature [F(7, 490) = 61.34, P<0.001]. Cold stimuli PERIPHERY
produced a marked excitation in the majority of Vc cells
(30/34). In the Vc, mean firing rates decreased sharply Long-term acclimation to capsaicin solutions affects taste
from 109.9 to −49.2 Hz with oral warming. In NTS, 76% bud volume and consumption in rats
of cells (29/38) were influenced by temperature; 9 were Jacquelyn M Omelian, Suzanne I Sollars
excited by cold, 15 excited by warmth, and 5 responded
University of Nebraska at Omaha/Psychology Omaha,
to both kinds of stimuli. Thus, excitation to oral warming
NE, USA
was more prevalent in NTS. Results suggest Vc and NTS
process oral temperature information in distinct manners. The effects of chronic exposure to capsaicin (the component
Acknowledgements: NIH DC 011579 (CHL) responsible for the piquancy of chili peppers) in the gustatory
system are not yet well understood; a critical consideration that all of the tested compounds except tetradecane acti-
given capsaicin’s popularity in culinary and medicinal vate the rat trigeminal nerve. Then, we examined behavioral
applications. To examine these potential effects, rats received aversion responses to these compounds by placing a rat in
40 days of treatment with a 30% sucrose solution containing a square plexiglass arena with petri dishes in each corner.
either a 5 ppm capsaicin (in 2.5% ethanol) or sham (2.5% After a period of habituation, an insect chemical defense
ethanol) condition. Animals began exposure as neonates compound was added to one of the four petri dishes and
(P5) or adults (P40) to evaluate potential differences across the movement of the rat recorded. Ethovision XT (Noldus)
development. Taste bud volumes within fungiform papillae was used to analyze whether rats spent less time in the irri-
were measured at either two or 50 days post treatment, to assess tant containing corner. Rats were behaviorally averse to
immediate or lasting effects. Animals treated with capsaicin most compounds at concentrations similar to those released
as neonates had significantly smaller taste buds at 50 days by insects. By imaging primary cultures of rat trigeminal
post treatment but in no other group (neonate or adult) were ganglia using a calcium responsive dye, we have confirmed
there significant differences. Capsaicin is a trigeminal irritant the activation of the trigeminal system by insect chemical
and does not directly affect the chorda tympani-innervated defense compounds. Additional experiments are underway
taste buds, thus the difference in taste bud volumes following to determine the specific receptor targets of these chemi-
capsaicin exposure suggests an integrated relationship cals using a heterologous expression system. In the future,
between the chorda tympani and lingual nerves in gustatory we will be doing similar work with chickens to determine
maintenance. The capsaicin concentrations used here were the role of the trigeminal system in avian responses to insect
significantly lower than those found in nature, as treatments defense compounds. Acknowledgements: WFU Center for
were limited to what animals would consume willingly. As Molecular Communication and Signaling
an additional experiment, we examined whether rats develop
a tolerance to capsaicin over time. To do so we gave adult
animals 5 ppm capsaicin/30% sucrose solutions and then #P120 POSTER SESSION III: TRIGEMINAL;
incrementally increased the quantity of capsaicin by 2.5 HUMAN OLFACTORY PSYCHOPHYSICS; TASTE
ppm after each 5 day period. Initial results showed animals PERIPHERY
willingly consumed higher levels of capsaicin (10 ppm) with
this type of acclimation. Taste bud volume analyses for these Cholingeric Neurotransmission Links Solitary Chemosensory
animals will be presented, and further experiments with Cells To Nasal Inflammation
additional acclimation are ongoing. Acknowledgements: CJ Saunders1,2, Thomas E Finger1,2, Marco Tizzano1
University of Nebraska at Omaha: Graduate Research and 1
Rocky Mtn Taste & Smell Center, Univ Colo School of
Creative Activities Fund
Medicine Aurora, CO, USA, 2Neuroscience Program, Univ
Colo School of Medicine Aurora, CO, USA
#P119 POSTER SESSION III: TRIGEMINAL; Solitary chemosensory cells (SCCs) are specialized epithelial
HUMAN OLFACTORY PSYCHOPHYSICS; TASTE chemosensors that respond to “bitter” substances via the
PERIPHERY canonical taste transduction cascade (T2Rs, Gα-gustducin
and TRPM5). When stimulated, SCCs release a hitherto
Trigeminal Mediation of Mammalian Aversion to Insect unidentified neurotransmitter onto peptidergic nociceptive
Chemical Defense Compounds trigeminal fibers. Activation of these nociceptive fibers
Paige Richards, Pamela Fazio, Alexa Ciesinski, Annalyn triggers neurogenic inflammation via NK1 (substance P)
Welp, Deirdre Craven, Wayne Silver receptors on capillaries causing local plasma extravasation
(Tizzano & Finger, AChemS 2012). How SCCs activate
Wake Forest University Winston-Salem, NC, USA
nerve fibers is unknown. In the present study, we show
Insects release defensive chemicals as one mechanism for that choline acetyltransferase, the synthetic enzyme for
deterring predation and defending their territory. A wide acetylcholine (ACh), is present in SCCs. Previous studies
variety of insects use the same chemicals for this purpose. have shown nicotinic ACh receptors (nAChR) on trigeminal
We hypothesize that defensive insect chemicals used by mul- fibers. Thus, all elements for cholinergic neurotransmission
tiple insect orders are targeting the mammalian trigeminal are present in the SCC-trigeminal system. To test if SCC-
system and eliciting chemesthesis. In the current study, we mediated inflammation requires activation of nAChRs,
test a number of insect defense compounds to determine we measured SCC-evoked plasma extravasation in mice
if they are irritating to a mammalian predator (rat). We stimulated unilaterally with denatonium benzoate (20 µL,
first determined if these defense compounds activated the 10mM). Prior to chemical stimulation, mice were injected i.p.
trigeminal nerve by recording from the ethmoid branch with either saline or the nAChR-antagonist Mecamylamine
and monitoring respiration while perfusing stimuli through (Mec). Under urethane anesthesia, the right naris was
the rat’s nasal cavity. Using these methods, we determined stimulated with denatonium, and the mouse injected i.v. with
Alexa555- conjugated albumin. Heads were bisected and #P122 POSTER SESSION III: TRIGEMINAL;
fluorescence intensity in the nasal epithelium was quantified. HUMAN OLFACTORY PSYCHOPHYSICS; TASTE
A one-way ANOVA demonstrated a significant difference PERIPHERY
between the three experimental groups [F(2,12)=17.03;
p<0.001], with the saline control showing significantly more The Perception of Novel Cooling Ingredients in Flavored
extravasation than either the 3mg/kg Mec [p<0.01] and Beverages Varies with Ethnicity and Prior Experience
6 mg/kg Mec [p<0.001] groups (Tukey’s HSD test). This Xaiorong Su1, Beverly J Tepper1, Jennifer B Tartaglia2, Carter
result supports the hypothesis that SCCs are cholinergic, like Green2
tracheal brush cells (Krasteva, PNAS 2011), and release ACh
Rutgers University/Food Science New Brunswick, NJ, USA,
1
to activate nAChRs on trigeminal fibers when stimulated.
Takasago International Corporation (U.S.A.) Rockleigh, NJ,
2
Acknowledgements: NIDCD R03 DC012413 (M.T.), R01
USA
DC009820 (T.E.F.), and P30 DC04657 (to D. Restrepo)
Derivatives of l-menthol are cooling ingredients that are
#P121 POSTER SESSION III: TRIGEMINAL; widely used in confectionery and oral care, and may have
HUMAN OLFACTORY PSYCHOPHYSICS; TASTE applications in beverages. To our knowledge, no studies
PERIPHERY have examined ethnic differences and prior experience with
cooling ingredients in the sensory perception of cooling in
Both painless and dTRPA1, Drosophila TRPA Channels, model beverages. This study examined the oral sensations
Detect Chemical Irritants of cooling, heat/burning, tingling, sweetness, bitterness
Wayne L Silver, Madison L Shoaf, Pam A Fazio, Erik C and overall flavor from lemon-lime flavored beverages
Johnson with two novel cooling ingredient blends; Coolact® 38D/
(l)-menthyl lactate blend or Coolact® 5 /Coolact®10 blend
Wake Forest University/Biology Winston-Salem, NC, USA in healthy adults who were East Asian (n=54), Caucasian
The ability to detect chemical irritants is important for the (n=43) or other (n=19). Subjects rated intensity and liking
avoidance of potentially life threatening compounds. In of each blend at four concentrations (0, 75, 150, and 300
mammals, one target of these irritants is the TRPA1 chan- ppm) using 15-cm line scales at four time points (0, 2.5, 5
nel. The fruit fly, Drosophila, has four homologs of mam- and 10 min) after tasting. The intensity of all attributes was
malian TRPA1, two of which are painless and dTRPA1. maximal immediately after tasting (P<0.0001) and decreased
Previous research has provided contradictory evidence with time (p<0.0001). Both blends primarily delivered the
about the roles of painless and dTRPA1 in fly chemical noci- sensations of cooling and tingling with minimal perception
ception. We used the proboscis extension reflex and devel- of heat/burning and bitterness. At time 0, East Asians
oped a two-bottle preference test to analyze the behavioral perceived more heat/burning than Caucasians from
phenotypes of painless and dTRPA1 mutants. Both assays Coolact®5/Coolact®10 blend (p<0.01). Also, subjects who
indicate that each channel is required for the behavioral were familiar with flavored beverages containing cooling
aversion to AITC, though it is not clear whether they are ingredients (n=60) perceived more cooling, heat/burning and
acting independently or in combination. We evaluated the tingling from the Coolact®5/Coolact®10 blend (p<0.0001)
expression patterns of painless and dTRPA1 to determine and more cooling (p<0.001) and heat/burning (p<0.01) from
if there was colocalization. No overlap was seen centrally, the Coolact® 38D/(l)-menthyl lactate blend compared to
and we are currently evaluating colocalization peripherally. subjects who were not familiar with these products (n=56).
To further define these cell populations, specific cell mark- These data suggest that the intensity of cooling ingredients
ers were identified. We observed subsets of painless and in beverages is influenced by ethnicity and prior experience
dTRPA1 that were colocalized with the fly homologs of with these beverages.These factors should be considered in
mammalian CGRP and Substance P, respectively. Cell excit- future psychophysical studies of cooling ingredients and
ability of painless and dTRPA1 cells was assessed by using product applications Acknowledgements: Supported by
a Ca+2 reporter, GCaMP, to observe changes in Ca+2 levels Takasago International Corporation (U.S.A.).
in response to AITC. Both painless and dTRPA1-expressing
cells displayed significant increases in fluorescence following
application of AITC. To determine if activation occurred #P123 POSTER SESSION III: TRIGEMINAL;
directly or indirectly, painless and dTRPA1 were expressed HUMAN OLFACTORY PSYCHOPHYSICS; TASTE
in an ectopic Drosophila tissue. Again, both painless and PERIPHERY
dTRPA1 demonstrated increases in cell excitability to AITC,
Peptidergic Trigeminal Nerve Fibers are Required for
suggesting that these channels are acting independently to
Solitary Chemosensory Cell-mediated inflammation
detect irritants. Collectively, our results suggest a complex
following chemical insult of the nasal mucosa
neural circuit that requires both the painless and dTRPA1
channels for chemical nociception. Marco Tizzano1, Michael Christensen1,2, Thomas E. Finger1
1
Rocky Mtn Taste & Smell Ctr, Dept. Cell Developmental fact required to perceive carbonation bite. Subjects rated oral
Biology, Univ. of Colorado School of Medicine Aurora, CO, USA, pungency from several concentrations of carbonated water
2
Department of Bioscience, Aarhus University Aarhus, Denmark both at normal atmospheric pressure (at which bubbles could
The nasal epithelium houses a population of solitary chem- form) and at 2.0 atmospheres pressure (at which bubbles did
osensory cells (SCCs) that express T2R taste receptors along not form). Ratings of carbonation bite under the two pressure
with their downstream signaling components and which conditions were essentially identical, indicating that bubbles
are heavily innervated by peptidergic (substance P) trigemi- are not required for pungency. In Experiment 2, we created
nal nerve fibers. Nasal SCCs respond to bitter compounds controlled streams of air bubbles around the tongue in mildly
including bacterially-produced molecules, to evoke protec- pungent CO2 solutions to determine how tactile stimulation
tive respiratory reflexes and early inflammatory responses from bubbles affects carbonation bite. Since innocuous sen-
(Gulbransen 2008, Tizzano 2010 and AChemS 2012). Here sations like light touch and cooling often suppress pain, we
we test whether SCC- mediated pro-inflammatory responses predicted that bubbles might reduce rated bite. Contrary to
require activation of the trigeminal nerve and subsequent prediction, air bubbles flowing around the tongue signifi-
peptidergic neurotransmission, or whether SCC activation cantly enhanced rated bite, without inducing perceived bite
triggers local inflammation via an intramucosal paracrine in blank (un-carbonated) solutions. Accordingly, though
signaling mechanism. When denatonium (10mM) is instilled bubbles are clearly not required for carbonation bite, they
into the nasal passageways, it evokes SCC-dependent plasma may well modulate perceived bite. More generally, the results
leakage and local mast cell (MC) degranulation (Tizzano show that innocuous tactile stimulation can enhance chemo-
AChemS 2012). Chemical ablation of peptidergic nerve genic pain. Possible physiological mechanisms are discussed.
fibers with resiniferatoxin (RTX, an ultrapotent analog of Acknowledgements: Supported in part by Anheuser-Busch
capsaicin) eliminates both denatonium- mediated plasma InBev
leakage and MC degranulation. These results show that
the peptidergic nerve fibers are necessary for these SCC-
mediated pro-inflammatory events. Moreover, injection of #P125 POSTER SESSION III: TRIGEMINAL;
L732138 (5mg/kg), an inhibitor of the neurokinin 1 (sub- HUMAN OLFACTORY PSYCHOPHYSICS; TASTE
stance P) receptor present on blood vessels, prevents denato- PERIPHERY
nium- induced plasma leakage. This indicates that substance Acid detection by TRPV1 channels in both ‘taste blind’
P is the mediator for plasma leakage. Our results demon- (P2X-KO) and C57 mice
strate that activation of the SCCs leads to a rapid, local
pro-inflammatory response via neurogenic mechanisms. Meghan L Bills, Jennifer M Stratford,
This fast pro-inflammatory response, driven by the SCCs in Thomas E Finger University of Colorado School of Medicine/
conjunction with the trigeminal nerve, represents a 1st line Department of Cell and Developmental Biology Aurora, CO,
of defense against respiratory epithelial assault by noxious USA
chemicals and bacterial pathogens. Acknowledgements:
NIDCD R03 DC012413 (M.T.), R01 DC009820 (T.E.F.), In the oropharynx the low pH of acidic solutions is
and P30 DC04657 (to D. Restrepo) detected via taste as sour, and by general mucosal fibers
as pungency or chemesthesis. This results in the behavioral
avoidance of acidic stimuli but the relative contribution
#P124 POSTER SESSION III: TRIGEMINAL; of these two systems is unknown. Genetic deletion of the
HUMAN OLFACTORY PSYCHOPHYSICS; TASTE purinergic receptors P2X2 and P2X3 (P2X-KO) results
PERIPHERY in interruption of taste bud-to- nerve transmission and
consequent loss of taste responses in the gustatory nerves.
The Influence of Bubbles on the Perception of Although P2X-KO mice do not respond behaviorally
Carbonation Bite to most tastants, they continue to avoid acids at similar
Paul M Wise1, Stephen R Thom2, Madeline Wolf1, Bruce concentrations as wildtypes. General mucosal afferents
Bryant1 express TRPV1 channels, which are activated by low pH
and may underlie this avoidance of acids. To test this,
1
P2X-KO (n=8) and C57 (n=12) mice were assessed using
University of Pennsylvania/Institute for Environmental
2
a two-bottle preference test in which one bottle contained
Medicine Philadelphia, PA, USA
20 mM citric acid (CA) and the other water. The test was
Although many people assume that the bite of carbona- given in the presence and absence of the TRPV1 antagonist
tion is due to tactile stimulation of the oral cavity by bub- Iodoresiniferatoxin (I-RTX). Both strains showed a
bles, it has become increasingly clear that carbonation bite significant decrease in avoidance with I-RTX versus vehicle
comes mainly from formation of carbonic acid in the oral (p<0.05) indicating a role for TRPV1 in acid avoidance.
mucosa. In Experiment 1, we asked whether bubbles were in Additionally, C57 mice that had been injected repeatedly
with Resiniferatoxin (RTX) to eliminate TRPV1 nerve Ca2+ influx in hTRPA1-exrpessing cells in the same man-
terminals were tested using a two-bottle preference test ner as in hTRPV1 with an EC50 value of 57.2 μM. The
with 20 mM CA and water. The RTX injected mice had a NGCC-induced Ca2+ influx was blocked by ruthenium
higher acid intake, 19%, than controls, 11%. This difference red (30 μM), and HC-030031 (100 μM), a specific antago-
was similar to the change in acid intake of C57 mice given nist of TRPA1. These data provides evidence that NGCC
I-RTX, 21%, versus vehicle, 12%. These results indicate selectively activate TRPV1 and TRPA1 in cultured cells.
that the TRPV1 channel is important in non-taste acid These data further support our previous suggestion that
avoidance. However, P2X-KO mice still avoided CA even NGCC interact with the TRPV1 variant cation channel
with the TRPV1 channel blocked suggesting that TRPV1 in the anterior taste receptive field. Acknowledgements:
does not account for all non-taste acid detection and that Supported by a Korea Food Research Institute (KFRI)
other mechanisms are likely involved, e.g. TRPA1 (Wang grant E0121201 and DC-011569
et al. 2011). Acknowledgements: Supported by NIH Grant
R56DC000147 (TEF) and P30 DC04657 (Rocky Mountain
Taste & Smell Center). #P127 POSTER SESSION III: TRIGEMINAL;
PERIPHERY
#P126 POSTER SESSION III: TRIGEMINAL; Olfactory Overshadowing: The Effect of Verbal and Tactile
HUMAN OLFACTORY PSYCHOPHYSICS; TASTE Stimuli on Olfactory Memory
PERIPHERY Nicole K Beers, Amy E Callaham, David E Hornung
N-geranylcyclopropylcaboximide (NGCC) selectively activate Biology Dept. St. Lawrence University Canton, NY, USA
hTRPV1 and hTRPA1 in cultured cells
While the strong association between smell and memory
MR Rhyu1, HJ Son 1, Y Kim1, MJ Kim1, SH Song1, MJ Cheong has received considerable attention, little is understood
M1, T Misaka2, M.L. Dewis3, V Lyall4 about the effect non-olfactory stimuli have on the encoding
of olfactory memory. In the “verbal overshadowing”
1
Korea Food Research Institute Seongnam-Si, South Korea,
part of this study, subjects were divided into 5 groups
2
The University of Tokyo Tokyo, Japan, 3International
which differed in the type of verbal response attached
Flavors & Fragrances Union Beach, NJ, USA, 4Virginia
to 10 target odors presented during a training phase. In
Commonwealth University Richmond, VA, USA
the 4 experimental groups, subjects verbalized names or
In mammals, two salt taste pathways have been character- descriptors whereas subjects in the control group provided
ized: one is selectively responsive to Na+, which is inhib- no verbal name or description. After participating in an
ited by amiloride and the other is Na+ non-specific and unrelated task, subjects were presented with a battery of
amiloride- insensitive. As the amiloride-sensitive Na+ odorants one at a time (10 targets and 10 distracters) and
specific salt taste receptor, the epithelial sodium chan- asked if they remembered smelling each odorant. Subjects
nel (ENaC) has been validated. The transient receptor who verbalized a name or description recognized fewer
potential vanilloid-1 variant salt taste receptor (TRPV1t) odorants as compared to subjects who provided no verbal
has been proposed as a constitutively active non-selective response. In addition, the subjects who provided no verbal
cation channel that has many similarities with the pain response identified fewer distracter odorants as belonging to
receptor TRPV1. In previous report we have shown that the target group. In the “temperature overshadowing” part
NGCC synthesized by IFF, modulates salt taste on human of this study, subjects had cold or room temperature water
and amiloride-insensitive NaCl chorda tympani taste applied to their hands while they concurrently smelled the
nerve responses by interacting with TRPV1t. In this pres- target odors. Subjects who had water applied to their hands
entation, we performed calcium imaging and cell based during the training phase recognized fewer of the target
assay using in hTRPV1-expressing cells to test the inter- odorants as compared to subjects in a control group from
action with NGCC and TRPV1NGCC enhanced Ca2+ the verbal overshadowing part of the study who had no
influx in hTRPV1-exrpessing cells in a time- and dose- water applied to their hands; additionally subjects recalled
dependent manner with an EC50 value of 98.7 μM. The fewer of the odors that were paired with the cold stimulus
NGCC-induced Ca2+ influx was markedly attenuated by than those that were paired with room temperature water.
ruthenium red (30 μM), a general blocker of TRP chan- The data from both parts of this study is consistent with
nels, and capsazepine (5 μM), a specific antagonist of the hypothesis that olfactory memory is impaired when
TRPV1, implying NGCC directly activate TRPV1. On the target odorants are coupled with verbal or tactile stimuli.
other hand, TRPA1 is often co-expressed with TRPV1 in Acknowledgements: The St. Lawrence University Fellows
sensory neurons therefore we investigated the effects of Program and the Biology Department provided some of
NGCC on hTRPA1-expressing cells. NGCC enhanced the funding for this work.
P128 POSTER SESSION III: TRIGEMINAL; Kingdom, 3International Flavors & Fragrances Union Beach,
HUMAN OLFACTORY PSYCHOPHYSICS; TASTE NJ, USA
PERIPHERY Olfactory research offers few trustworthy odor thresholds.
A new paradigm for testing olfactory memory performance Bad for the field, the situation sets nonexperts in need of
in healthy humans the information adrift. A rule of thumb says to choose
the lowest threshold of a set because studies with the best
Yvonne F. Brünner1, Christian Benedict2, Jessica Freiherr1 stimulus control and some measure of concentration
Diagnostic and Interventional Neuroradiology Aachen,
1 normally yield the lowest values. Nagata et al. over years
Germany, 2Institute of Neuroscience Uppsala, Sweden produced the largest set of thresholds gathered with coherent
methodology and analytical verification. They lie below
Long-term memory processes can be divided into two cat-
most others. Abraham et al. used a well-established linear
egories: declarative and procedural memory. While declara-
solvation energy relationship (LSER) to describe the set. To
tive memory concerns actual knowledge of facts, procedural
strengthen the position, they incorporated sets from the CPL
memory applies to more unconscious memory of abilities
and Hellman. Data obtained via olfactometer in recent years
and skills. Declarative memory processes are based upon the
from the CPL needed no normalization with the Nagata set,
two components recollection and recognition. Odors are con-
whereas those from Hellman and earlier squeeze- bottle
sidered highly salient but abstract evocative cues during the
data from the CPL did. The LSER accommodated these via
recollection of past personal experiences and events. The aim
indicator variables (factors) to bring the data into line and
of our current study was to develop a paradigm for testing
described potency for 353 odorants. Although the equation
declarative memory processes with regard to odors. Therefore,
left about 25% of variance unaccounted for, it offers the
we implemented three memory tasks in E-prime 2.0 presenta-
strongest statement yet for prediction. Until now, no sets
tion software. With help of an odor-place associative memory
included fragrance materials, such as patchouli oil or vanillin,
task we investigated memory of odor-place combinations.
known for potency. Without them, one could argue that the
During an odor item recognition task the subjects were asked
equation might describe local rather than universal rules. We
if they had experienced several odors during the previous task.
accordingly ran groups of at least 10 Ss on over a dozen such
As a control task for the first task a picture-place associative
materials. We used the CPL protocols that have given stable
memory task was utilized. Each of those tasks contained two
results and required no normalization. The outcome fell in
phases – an encoding phase (1 block) and a retrieval phase (4
line with the data for the hundreds of materials, with neither
blocks) – and were applied to 17 healthy subjects. The results
more nor less variance. It adds evidence that the LSER has
suggest that the subjects were able to learn during the encod-
universal relevance to predict potency. Since the solvation-
ing as well as during the retrieval phase. We found higher
relevant parameters used for prediction exist for thousands
odor-place associative, odor recognition, and picture-place
of materials, the LSER may afford more accessibility and
associative memory performance scores at the end compared
accuracy than values in compilations. Acknowledgements:
to the beginning of the retrieval phase. We thus claim that our
Supported by International Flavors and Fragrances.
paradigm renders useful during the investigation of olfactory
memory processes and the influence of external factors on
those processes in humans. We here present a fully automated #P130 POSTER SESSION III: TRIGEMINAL;
paradigm, which can be utilized during future functional imag- HUMAN OLFACTORY PSYCHOPHYSICS; TASTE
ing studies with the goal to shed light onto the neural network PERIPHERY
of human olfactory memory processes. Acknowledgements:
This research was funded by a startup grant from the Medical Categorical Dimensions of Human Odor Descriptor Space
Faculty of RWTH Aachen University. Revealed by Non-Negative Matrix Factorization
Jason B Castro1, Arvind Ramanathan2, Chakra S Chennubhotla3
#P129 POSTER SESSION III: TRIGEMINAL; 1
Bates College,Department of Psychology, Program in
HUMAN OLFACTORY PSYCHOPHYSICS; TASTE Neuroscience Lewiston, ME, USA, 2Oak Ridge National
PERIPHERY Laboratory Oak Ridge, TN, USA, 3University of Pittsburgh,
Department of Computational and Systems Biology
Fragrance Materials Extend the Range of a Model for Odor Pittsburgh, PA, USA
Potency
Recent studies using Principal Components Analysis
William S Cain1, Roland Schmidt1, Jorge E Cometto-Muñiz1,
(PCA) support low-dimensional models of odor space, in
Sang Park1, Craig B Warren1, Michael H Abraham2, Matthias
which one or two dimensions – with hedonic valence fea-
Tabert3
turing prominently – explain most odor variability. Here
1
Chemosensory Perception Lab (CPL) La Jolla, CA, USA, we use non-negative matrix factorization (NMF) – a non-
2
University College London / Chemistry London, United linear optimization method - to discover an alternative,
reduced-dimensional representation of the Dravnieks odor best-fitting genetic models showed that over 30% of the vari-
database(144 odors x 146 descriptors). We first divided ances in binaral rivalry rate and binaral rivalry magnitude,
the dataset into training and testing halves, and found respectively, were accounted for by additive genetic factors.
that RMSD testing error attained a minimum for sub- Our study represents the first large sample study of binaral
space choice of 25, motivating this as an upper bound for rivalry. The findings demonstrate a reliable genetic compo-
odor perceptual space dimensionality. More parsimonious nent of this phenomenon and suggest an innate rhythm of
representations were found by comparing reconstruction olfactory perception.
errors (fraction of unexplained variance) of NMF with
reconstruction errors of PCA on scrambled data (PCAsd).
For subspace sizes > 10, NMF error was indistinguishable
from PCAsd error, indicating no gain in retaining more #P132 POSTER SESSION III: TRIGEMINAL;
than 10 perceptual dimensions. As is typical of NMF basis HUMAN OLFACTORY PSYCHOPHYSICS; TASTE
sets, the 10 odor dimensions we obtain are sparse (only a PERIPHERY
small subset of the 146 descriptors apply), and categorical Olfactory Plasticity in Young Adults
(represent a positive valued quality). Moreover, these 10
dimensions were near- orthogonal, with a mean angle of Beverly J. Cowart1, Marcia L. Pelchat1, Johan
73 degrees between all pairs of basis vectors. Investigating N. Lundström1,2,3, Ryan Crawford1, Lydia Milbury1, Kathrin
the distribution of odors in this 10-dimensional space, we Ohla1,4
find marked clustering, with each odor clearly defined by 1
Monell Chemical Senses Center Philadelphia, PA,
its membership in a single dimension, to the exclusion of USA, 2Dept. of Psychology, University of Pennsylvania
others. Members of each cluster have notable structural Philadelphia, PA, USA, 3Dept. of Clinical Neuroscience,
homology, which we quantified as correlations among Karolinska Institute Stockholm, Sweden, 4German Institute
physiochemical descriptors of odors in each cluster. In of Human Nutrition Potsdam-Rehbrücke Nuthetal,
sum, we describe a representation of odor perceptual Germany
space consisting of 10 discretely occupied dimensions that
apply categorically. We propose that this may help eluci- There is considerable evidence that olfaction is, in many
date the natural axes of olfaction. Acknowledgements: ways, a “learned” sense, showing experience-induced
NIH GM086238 (CSC) plasticity in both central circuits and peripheral receptors
even in adulthood. Although this has been demonstrated in
humans, the focus has been on single chemicals to which some
#P131 POSTER SESSION III: TRIGEMINAL; individuals are initially insensitive. We sought to explore
HUMAN OLFACTORY PSYCHOPHYSICS; TASTE (1) the possibility of enhancing young adult sensitivity to
PERIPHERY complex odors, (2) the role of cognitive engagement (active
identification of exposed odors vs. simple exposure), and
Genetic Contribution to Binaral Rivalry (3) the extent of transfer of learning to unexposed odors.
Kepu Chen, Bin Zhou, Guo Feng, Wen Zhou For this, we obtained both odor thresholds and olfactory
event-related potentials (OERPs) from 40 young adults in
Institute of Psychology, Chinese Academy of Sciences
an attempt to assess the neuronal mechanisms of potential
Beijing, China
behavioral changes. Thresholds for four exposed and two
When two odorants of different structure and smell are unexposed complex odorants were obtained at baseline, 6
simultaneously presented to the two nostrils, we experi- and 12 weeks; OERPs in response to two of the exposed
ence alternations in olfactory percepts, a recently discovered and both unexposed odorants were obtained at baseline and
phenomenon termed binaral rivalry. In an effort to further 12 weeks. Our behavioral results suggest that intermittent
characterize this phenomenon and its nature, we adopted odor exposure in these circumstances does enhance
the twin method and tested monozygotic (MZ) twins (n = 73 threshold sensitivity and that cognitive engagement may
pairs) that are genetically identical and dizygotic (DZ) twins further enhance generalization to unexposed odorants.
(n = 70 pairs) that share about half of their genes. The Our electrophysiological results show learning-dependent
majority of participants experienced binaral rivalry over a amplitude changes, particularly of the late positive
course of 20 samplings of eugenol and amyl acetate, one component. Taken together, these data provide further
to each nostril. Large variances are observed in the number support for the notion that repeated exposure augments
and the magnitude of their perceptual switches. Critically, olfactory sensitivity and that cognitive mechanisms exert
such individual differences are partially genetic. The cor- a significant modulatory effect. Acknowledgements:
relations between MZ twins for the two aforementioned Supported by the U.S. Army Research Office, grant
indexes are both higher than those between DZ twins. The #W911NF-11-1-0087.
#P133 POSTER SESSION III: TRIGEMINAL; #P134 POSTER SESSION III: TRIGEMINAL;
HUMAN OLFACTORY PSYCHOPHYSICS; TASTE HUMAN OLFACTORY PSYCHOPHYSICS; TASTE
PERIPHERY PERIPHERY
Proton-Transfer-Reaction Mass Spectrometry Effects of Cinnamon Scent Administration on Physiology,
Range of Motion, Mood, Anxiety and Perceived Workload
Melanie Y Denzer1, Jonathan Beauchamp2, David W Kern3,
During a Multi-session Physical Therapy Program
Stefan Gailer2, Norbert Thuerauf4, Johannes Kornhuber4,
Andrea Buettner1,2 Jessica Florian, Kristen Johnson, Sierra Moore, Bryan
Raudenbush, Allison Burke
1
Department of Chemistry and Pharmacy, Emil Fischer
Center, University of Erlangen-Nuremberg Erlangen, Wheeling Jesuit University Wheeling, WV, USA
Germany, 2Department of Sensory Analytics, Fraunhofer
Scents have been shown to elicit both emotional and
Institute for Process Engineering and Packaging IVV
physiological responses. The current study aimed to evaluate
Freising, Germany, 3Department of Comparative Human
the possible effects of cinnamon scent when applied to a
Development, Institute for Mind and Biology, University
physical therapy regimen. Forty-two undergraduate students,
of Chicago Chicago, IL, USA, 4Department of Psychiatry
16 males and 26 female, completed a four trial physical
and Psychotherapy, University of Erlangen Erlangen,
therapy regimen in one of two rooms: a control room or
Germany
a room infused with a cinnamon scent. The experimenters
Since their introduction in the mid-1990s, Sniffin’ Sticks measured participants’ range of motion, mood (POMS),
have been used effectively by many otolaryngologists to and anxiety (STAI) prior to and following each trial of
assess olfactory dysfunction in countless patients. Despite exercises. At the end of each visit, perceived workload was
their widespread use, however, there is currently a lack of assessed (NASA-TLX). The data were analyzed using a 4
data on the actual odorant concentrations released from the (visits) X 2 (groups) mixed design ANOVA. Significant
tips of these pens and whether these emitted concentrations results were found for ratings of effort on the NASA- TLX,
scale linearly in accordance with the odorant concentrations F(3, 120)=2.8, p = .042. Participants in the cinnamon scent
of the pen set. The purpose of this study was to ascertain condition rated their perceived effort exertion as being
whether the Sniffin’ Sticks’ presumed odorant release was lower than participants in the control condition. Decreased
concordant with the concentration of the odorant solu- perceived effort may cause patients undergoing a physical
tions placed in the pens. The commercially-available odour therapy program to feel more comfortable while completing
threshold test containing n-butanol was chosen here for their exercises, thus increasing the likelihood of adherence
evaluation. The threshold set contains concentrations (v/v) to the program.
ranging from 4 % (pen no. 1) to 1.2 ppm (pen no. 16), with
stepwise 1:2 dilutions. We also tested an additional custom- #P135 POSTER SESSION III: TRIGEMINAL;
made pen containing 8 % n-butanol (pen no. 0). The odor- HUMAN OLFACTORY PSYCHOPHYSICS; TASTE
ant concentration emanating from the tip of each pen was PERIPHERY
measured directly via proton-transfer-reaction mass spec-
trometry (PTR-MS), which is an on-line analytical tool for Towards A Novel Method for Human Olfactory Research
detection and quantitation of volatile organic compounds Jessica M Gaby Vivian Zayas
(VOCs) – including odorants – at trace concentrations. The
Cornell University Ithaca, NY, USA
pens were also subjected to repeated use to ascertain the
degree of reproducibility of emitted odorant concentrations The majority of human olfactory research to date has been
under stress. These measurements showed that the concen- conducted without regard to perfume or dietary contribu-
tration linearity of n-butanol emitted over the range of pens tions to body odor. Given that humans have used perfume for
was excellent and highly reproducible. The stress tests dem- thousands of years, and that culturally mediated food pref-
onstrated that the emitted concentrations of n-butanol were erences may affect body odor, I propose that an ecologically
lower after repeated use of the pens compared to those of relevant model of olfactory communication should account
the unused pens, albeit with a mostly good linearity over the for body odor as people present themselves in real social situ-
entire range of pens. Acknowledgements: Part of this study ations. Further, current collection methods for odor samples
is affiliated with the Neurotrition Project, which is supported focus mainly on axillary secretions, though it is known that
by the FAU Emerging Fields Initiative. This work was also other body areas contribute to odor. This pilot study aims
partly supported by The National Social Life, Health and to validate a novel olfactory research method by employing
Aging Project Wave 2 (R37 AG030481). DWK is supported present others rather than disembodied odors. In a repeated
by The Center on Aging Specialized Training Program in the measures design, blindfolded, ear-plugged raters evaluated a
Demography and Economics of Aging, which is funded by series of participants as each sat beside them. No restrictions
the National Institute on Aging (NIA) (T32000243) in diet or fragrance were used, to mimic real-life interactions.
First and second ratings for each participant were examined potential roles of the medial prefrontal cortex during
for intertrial reliability, revealing significant consistency control versus prediction is currently underway and will
(p<.001), even though raters were unaware of our repeated also be discussed. Acknowledgements: The research in this
measures design. Self-report measures of odor sensitivity, presentation is supported by the National Institutes of Health
perfume usage, and perceived pleasantness of friends’ body [NIDCD 1 R01 DC 010014, NIMH 5 F32 MH 091967].
odors were significantly associated with pleasantness ratings
(p<.05). These preliminary results indicate reliable olfactory
#P137 POSTER SESSION III: TRIGEMINAL;
perception of body odor in this novel design. Upcoming iter-
ations will provide a larger sample size, and plans for future HUMAN OLFACTORY PSYCHOPHYSICS; TASTE
experiments include examining more typically purified body PERIPHERY
odor, as well as comparing purified to daily body odor, Pleasant and unpleasant odors are easier to detect and
which will allow us to gain insight into the potential effects harder to ignore
of fragrance and dietary choices on olfactory communica-
Jingwen Jin, Aprajita Mohanty
tion. Studies comparing lab samples to present others will
evaluate the generalizability of tightly controlled lab tech- State University of New York at Stony Brook/Department of
niques to real life situations. Acknowledgements: Institute Psychology Stony Brook, NY, USA
for the Social Sciences, Cornell University Odor objects are often experienced within the context of
several competing odors. To deal with the excess of informa-
tion the olfactory system utilizes mechanisms that bias the
#P136 POSTER SESSION III: TRIGEMINAL; competition between odors towards preferential represen-
HUMAN OLFACTORY PSYCHOPHYSICS; TASTE tation of the most relevant odor. This biasing may involve
PERIPHERY anticipatory attentional mechanisms that use feature-
Effects of Prediction and Control on the Perception of related information regarding behaviorally relevant odors to
Aversive Olfactory Stimuli amplify relevant odors and filter our irrelevant odors. Since
an important feature of olfactory stimuli is their valence, the
Katherina K. Hauner, Daniel T. Ohm, Jay A. Gottfried present study utilized an olfactory attentional search task to
Northwestern University Chicago, IL, USA examine whether 1) expectation of pleasant or unpleasant
odors enhances perceptual sensitivity compared to neutral
Prediction and control are distinct cognitive functions odors and 2) presence of an irrelevant pleasant or unpleas-
that have been shown to play significant roles in emotion ant odor make it harder to detect a relevant odor. Subjects
regulation. Animal studies have demonstrated that the ability performed an olfactory task in which they decided whether
to predict or exert control over the temporal onset of aversive a particular target smell is present in each trial. Trials started
stimulus presentation substantially decreases stimulus- with a cue that indicated the forthcoming target odor (pleas-
evoked physiological arousal. Whether these reductions in ant odor A, unpleasant odor B, or neutral odor C) followed
arousal are due to changes in the perception of the stimulus
by the target stimulus. The stimulus consisted of either odor
itself, however, is unknown. Here we examined the effects A alone, odor B alone, odor C alone, or binary mixture
of prediction and control on responses to aversive olfactory of these odors (AB, AC, and BC). In our results,d-prime,
stimuli in human subjects. The decision to use odorants a measure of perceptual sensitivity showed the following
as aversive stimuli was based on the intimate anatomical trend: pleasant odor following pleasant cues > unpleasant
and functional connections between olfactory and limbic odors following unpleasant cues > neutral odors following
systems—olfactory sensory neurons are only two synapses neutral cues (F(1,13)=7.240, p<0.05). Furthermore, accu-
removed from brain areas involved in emotion. Subjects were racy for detection of target odor was impaired when pre-
presented with aversive olfactory stimuli whose onset time sented in a mixture consisting of an unpleasant or pleasant
was either predictable, controllable, or neither. Responses to odor (F(1,13)=33.539, p<0.001). Present findings shed light
olfactory stimuli included self-report ratings of anticipatory on how emotional valence and selective attentional mecha-
anxiety and odorant aversiveness as well as stimulus-evoked nisms interact to impact the perception of olfactory objects.
physiological arousal (i.e., skin conductance response).
Critically, subjects received the same duration and frequency
of aversive stimulus delivery in each condition, and olfactory #P138 POSTER SESSION III: TRIGEMINAL;
stimuli were matched for valence and familiarity across all HUMAN OLFACTORY PSYCHOPHYSICS; TASTE
conditions. As hypothesized, results indicated that subjects’ PERIPHERY
responses to aversive olfactory stimuli differed depending on
Vanilla and Olfaction
whether stimuli were presented in a predictable or controllable
manner, despite no change in the frequency or duration of Joseph Wesley Jones, Hannah Tumlin, Jessica McKinney,
stimulus presentation. Follow-up fMRI research examining William Tewalt, Irene Nicky Ozbek
The University of Tennessee at Chattanooga Chattanooga, under endoscopic guidance. Participants did not require topi-
TN, USA cal anaesthetics or decongestants and sniffed naturally (n=35;
The purpose of this research is to establish norms for the odor- 21 female, 32 ± 7 years of age). An additional cannula fitted at
ant vanilla by using the University of Pennsylvania Smell the nostrils was connected to a pressure transducer to record
Identification Test (UPSIT), a norm referenced test of odor inspiratory nasal pressure and breathing cycles throughout
detection, as a template. Additionally, the odorants of banana odorant presentation. Following a protocol similar to that
and menthol, included on the UPSIT, were sampled to use as which is used during clinical assessments, n-butanol was pre-
a base for comparing relative equivalency of the norm sample. sented in a triad of Sniffin’ Sticks pens, 1 odorant target pen
Banana was chosen from amongst other UPSIT odorants due and 2 ‘odorless’ blanks. Participants were asked to identify the
to the somewhat similar sweet smell as compared to vanilla and odorant-containing target pen in each of six triads with increas-
menthol was chosen based on the involvement of the fifth cra- ing n-butanol concentrations (0.125, 0.25, 0.5, 2, 4, and 8 %).
nial nerve in processing this odorant. Vanilla was chosen due The entire protocol only required approximately 15 minutes to
to evidence that it may be preferred in infancy. Marchlewska- perform. All participants were fully compliant and reported
Koj, Lepri, and Muller-Schwarze (2001) found that premature only minimal discomfort. The catheter and cannula did not
infants showed a higher respiratory response to vanilla during disrupt normal breathing patterns. This protocol allowed us to
stimulation and no response to butyric acid. Savic-Burgland quickly and effectively measure odorant concentrations at the
(2004) using positron-emission tomographic scanning (PET olfactory cleft in real time. Comparing the known concentra-
scanning) showed that vanilla was processed differently. For tion of the stimulus odorant and the concentration found at
example, vanilla was found to influence only half of the olfac- the olfactory cleft will elucidate how to best adjust and develop
tory nerve. Results of the three odorants sampled by 380 of measures designed to determine an individual’s threshold sen-
subjects showed that banana and menthol had a correct identi- sitivity. Acknowledgements: This work was supported in part
fication rate of 59.7% and 88.9%, respectively, and vanilla had by The National Social Life, Health and Aging Project Wave
a 74.3% correct identification rate. The importance of includ- 2 (R37 AG030481), and is affiliated with the Neurotrition
ing vanilla as an odorant in smell identification tests used to Project, supported by the FAU Emerging Fields Initiative.
screen olfaction dysfunction is supported by this research. DWK is supported by The Center on Aging Specialized
Acknowledgements: Wheeler Center for Odor Research Training Program in the Demography and Economics of
Aging (National Institute on Aging (T32000243). TH received
funding from the DFG (HU 441/10–1).
PERIPHERY #P140 POSTER SESSION III: TRIGEMINAL;
Odorant Measurement at the Olfactory Cleft using Proton-
PERIPHERY
Transfer-Reaction Mass Spectrometry
Quantitative classification of the perceived intensity curve
David W. Kern1, Jonathan Beauchamp2, Mandy Scheibe3,
of a continuously presented odor
Thomas Hummel3, Martha K. McClintock1, Andrea Büttner2,4
Tatsu Kobayakawa1, Tomoko Matsubasa2, Naomi Gotow1
1
Department of Comparative Human Development, Institute
for Mind and Biology, University of Chicago Chicago, 1
The National Institute of Advanced Industrial Science and
IL, USA, 2Department of Sensory Analytics, Fraunhofer Technology (AIST) Ibaraki Tsukuba, Japan, 2Tokyo Gas Co.,
Institute for Process Engineering and Packaging IVV Ltd Tokyo, Japan
Freising, Germany, 3Smell & Taste Clinic, Department of Researchers of olfaction have devoted significant effort to
Otorhinolaryngology, Technical University of Dresden understanding the phenomenon of adaptation. Previous stud-
Dresden, Germany, 4Department of Chemistry and ies of olfactory adaptation have reported, in general, that per-
Pharmacy, Emil Fischer Center, University of Erlangen- ceived intensity of a continuously presented odor decreases
Nuremberg (FAU) Erlangen, Germany exponentially over time. More recently, however, other studies
Olfactory detection and threshold sensitivity is traditionally have shown that perceived intensity of continuous odor does
based on the concentration of the stimulus-odorant solution, not always decrease exponentially. Some studies focused on the
and does not typically consider the chemical concentration of various time-course of perceived intensity of a continuously
the stimulus-odorant at the level of the olfactory receptors. presented odor, whereas few studies have tried to classify time-
Here we report a technique for directly measuring odorant course patterns quantitatively. In this study, we performed
concentration in the olfactory cleft, utilizing Proton-Transfer- real-time evaluation of perceived intensity of a continuously
Reaction Mass Spectrometry (PTR-MS). PTR-MS samples presented odor for 480 seconds using nine odorants, and quan-
were collected using an approximately 15 cm long PVC cath- titatively classified the perceived intensity curves using the time
eter (outer diameter: 3.3 mm) placed at the olfactory cleft points for 20–80% of time integral of perceived intensity. This
analysis revealed that intensity curves could be classified into what precedes them; contextual effects. Hedonic contrast, is
two clusters: one group (“typical”) exhibited the commonly one of these phenomenon, when a stimulus is preceded by a
observed exponential decrease in perceived intensity; the other more pleasant stimulus, its pleasantness is rated lower (nega-
group (“non-typical”) included a range of time courses rather tive contrast), whereas when a stimulus is preceded by a less
than strict exponential decrease. The average intensity of pleasant stimulus, its pleasantness is rated higher (positive
“typical” group decreased below threshold three minutes after contrast). In this study, we investigated the contrast effect
beginning of odor exposure, and that of “non-typical” group, during evaluation of odor intensity and pleasantness. Two
on the other hand, did not decrease under threshold during sets of positive odors and a set of negative odors were used.
total measuring time. Group 1 sequentially rated the positive set A, the negative
set, and the positive set B. Group 2 rated the negative set
#P141 POSTER SESSION III: TRIGEMINAL; and then the positive set B. Group 3 rated only the positive
HUMAN OLFACTORY PSYCHOPHYSICS; TASTE set B. The mean rating of the negative and the positive set B
PERIPHERY were each compared among groups. As a result, in the inten-
sity rating, both positive and negative contrast occurred
Effects of Peppermint Scent Administration on Augmenting
clearly. In the pleasantness rating, only negative contrast
Cognitive and Creative Performance
was seen. This suggested that, in olfaction, the influence of
Lucas Lemasters, August Capiola, Bryan Raudenbush, the positive context stimuli on the following negative stimuli
Sierra Moore might be more robust. To investigate the cause of the non-
Wheeling Jesuit University Wheeling, WV, USA occurrence of positive contrast, we analyzed a change of
hedonic evaluation by rating order within a positive or nega-
Level of creativity has been assessed in a number of ways,
tive set. While there was no influence of the rating order in a
including interactions between body and environment on
negative set, the pleasantness rating of a positive set tended
creative thinking. Environmental richness has been shown to
to gradually decrease. This result is discussed in terms of
interact with creativity, with greater levels of environmental
the unstable characteristics for hedonic evaluation of posi-
richness leading to more creative responses. The present study
tive odors and the stability of the hedonic rating of negative
attempted to determine if peppermint scent administration
odors. Therefore, positive hedonic contrast may not occur
could promote creativity, since past research with peppermint
due to the hedonic value of negative odors being carried over
scent reports improved performance on clerical tests, thus
to the following rating of the better stimuli.
hinting at the possibility for cognitive enhancement. Participants
completed the Torrance® Tests of Creative Thinking, a
standardized test measuring creative thinking abilities, in
both a non-scented condition (control) and a peppermint #P143 POSTER SESSION III: TRIGEMINAL;
scented condition. Different versions of the test, as well as the HUMAN OLFACTORY PSYCHOPHYSICS; TASTE
conditions, were counterbalanced. The data were subjected to PERIPHERY
paired samples t-tests, with condition (peppermint, control) Increased cortisol secretion after awakening following
serving as the independent measure and raw scores of fluency, lavender inhalation during sleep
originality, elaboration, abstractness-of-titles, and resistance-
Shusaku Nomura1, Masako H. Ohira2, Toyonari Fujikawa1,
to-premature- closure serving as dependant measures. There
Kanetoshi Ito3
was a significant difference between the conditions for fluency
[t(33)=−2.41, p=.02], originality [t(33)=−2.13, p=.04], and 1
Nagaoka University of Technology/Engineering Nagaoka,
elaboration [t(33)=−7.38, p=.00], with all measures having Japan, 2Shiga University/Education Otsu, Japan, 3TAKASAGO
higher scores for the peppermint scent condition. Implications INTERNATIONAL CORPORATION Kanagawa, Japan
suggest working conditions for those individuals with Aromatherapy improves sleep through autonomous nervous
occupations that require creative thinking and problem solving system (ANS) modulation. However, few studies have focused
may benefit from peppermint scented working conditions. on the effect of olfactory exposure during sleep on hormonal
secretion. The objective of this study was to investigate the
effect of inhaling odorants on cortisol secretion during sleep
HUMAN OLFACTORY PSYCHOPHYSICS; TASTE and after awakening. Salivary cortisol was assessed because
PERIPHERY this glucocorticoid represents activation of the hypothalamus-
Contextual effects on hedonic evaluation of odors pituitary-adrenal (HPA) system, which is a dominant stress
reaction pathway in the body. We used essential oils of jasmine
Shiori Nakano Saho Ayabe-Kanamura
and lavender, which induce activation of the sympathetic and
Univ. of Tsukuba Tsukuba, Japan parasympathetic nervous systems (PSNS), respectively. These
When we sequentially evaluate a character of sensory modal- oils were administered to subjects through an olfactometer;
ity stimuli, evaluation of the stimuli might be influenced by volatilized odorants were intermittently delivered through a
cannula (first 1 min of each 5 min interval) during a 6-h sleep neural circuitry underlying aversive conditioning. These
period. Subjects experienced each odorant condition [jasmine, findings provide a framework for understanding perceptual
lavender, or scentless air (control) each night] and were exposed processing in anxiety disorders. Acknowledgements: This
to all three conditions in a counter-balanced order. Saliva material is based upon work supported by the U. S. Army
samples were collected every 30 min while the subjects were Research Office under grant number W911NF-11-1-0087
asleep using our own-developed saliva collection equipment
and every 15 min for 1 h after awakening. Surprisingly, salivary
cortisol after awakening was significantly higher under the #P145 POSTER SESSION III: TRIGEMINAL;
lavender condition than in the jasmine (p <.01) or control (p HUMAN OLFACTORY PSYCHOPHYSICS; TASTE
<.05) conditions, whereas no difference in cortisol secretion PERIPHERY
was observed while the subjects were asleep. Although lavender Perception of large and small odorant molecules – A study
has a sedative effect on humans thorough the activation of investigating the olfactory perception as a function of age
PSNS, inhaling this popular aroma during sleep may have a and odorant molecular size
significant impact on cortisol secretion by enhancing HPA
activity after awakening. Laura Puschmann, Charlotte Sinding, Thomas Hummel Smell
& Taste Clinic,
Department of Otorhinolaryngology, University of Dresden
#P144 POSTER SESSION III: TRIGEMINAL; Medical School Dresden, Germany
HUMAN OLFACTORY PSYCHOPHYSICS; TASTE For various sensory systems selective functional limitations
PERIPHERY with increasing age were detected. In vision and hearing it
Common aversive learning mechanisms to olfactory and shows in hyperopia or an increased hearing threshold for
visual stimuli high frequencies. But what about the olfactory system?
While some studies suggest a general olfactory impairment,
Valentina Parma1,2, Stacie S. Miller1,3, Fredrik Åhs4, Johan
others allow assumptions of partial processes. In this study
N. Lundström1,5,6
the influence of odorant molecular size was examined
1
Monell Chemical Senses Center Philadelphia, PA, USA, for the olfactory perception in different ages. Olfactory
2
Department of General Psychology, University of Padova threshold tests were conducted at two age groups (group
Padova, Italy, 3International Flavor and Fragrances Union 1: 18 to 30 years, group 2: 50 to 70 years). We used single
Beach, NJ, USA, 4Center for Cognitive Neuroscience, Duke odorants, bimolecular odorants and perfumes, each
University Durham, NC, USA, 5Department of Clinical consisting of large or small molecules. The results of these
Neuroscience, Karolinska Instituet Stockholm, Sweden, studies showed no differences for large and small odorant
6
Department of Psychology, University of Pennsylvania molecules in the young group of subjects. Whereas they
Philadelphia, PA, USA were perceived differently by the 50 to 70 year-old. The
Learning to avoid dangers is an evolutionary problem that latter had significantly higher thresholds for large olfactory
all organisms must solve. Aversive learning is a mechanism molecules. This phenomenon however, was detected only for
mediating the acquisition of conditioned responses (CR), mono- and bimolecular odorants. Perfumes were perceived
but it also alters the perception of the predictive cue, allow- similar in both groups. In Conclusion we can hypothesize
ing a differentiation of “threatening” and “safe” stimuli. It differing processes for odor perception of small and big
has been debated whether CRs to odors are stronger than odorant molecules in different age-groups at the receptor
to other sensory stimuli. We here investigated whether level. Moreover perfumes seem to generate more complex
increased conditioning to olfactory stimuli could be related olfactory information. The existence of partial processes
to an increased orienting response to odors due to their rela- on olfactory impairment with increasing age is expected for
tive sparseness compared to visual stimuli in our everyday the olfactory system, too. Acknowledgements: This project
environment. Twenty participants were tested employing was supported by a grant from DFG Schwerpunktprogramm
repeated conditioning (5 reinforcement sessions) of the same (SPP) 1392 - Integrative Analysis of Olfaction to Thomas
olfactory and visual stimuli over a period of two weeks and Hummel. In addition we thank Fragrance Resources GmbH,
compared to 20 matched individuals that only were exposed Hamburg for cooperation.
to one acquisition session. Although odor stimuli demon-
strated a larger initial CR than visual stimuli consistent with #P146 POSTER SESSION III: TRIGEMINAL;
an initially increased orienting response, this difference disap-
peared after repeated reinforcement sessions, suggesting the
presence of sensory independent aversive learning mecha- PERIPHERY
nisms. In conclusion, the strength of aversive conditioning is Odor Identification and Cognition in a Nationally
similar to olfactory and visual stimuli suggesting a common Representative Sample of Older Adults
L. Philip Schumm1, David W. Kern2, Kristen E. Wroblewski1, #P147 POSTER SESSION III: TRIGEMINAL;
Jayant M. Pinto3, Ashwin A. Kotwal4, William Dale4, Martha HUMAN OLFACTORY PSYCHOPHYSICS; TASTE
K. McClintock2 PERIPHERY
1
Department of Health Studies, University of Chicago Longitudinal Study of Olfactory Decline in a Nationally
Chicago, IL, USA, 2Comparative Human Development Representative Sample of Community Dwelling
and Institute for Mind and Biology, University of Chicago Older Adults
Chicago, IL, USA, 3Section of Otolaryngology-Head and
Neck Surgery, Department of Surgery, University of Chicago Kristen E Wroblewski1, Jayant M Pinto2, David W Kern3, L
Chicago, IL, USA, 4Department of Medicine, Section of Philip Schumm1, Martha K McClintock3
Geriatrics & Palliative Medicine, University of Chicago
Chicago, IL, USA
1
Department of Health Studies, University of Chicago
Chicago, IL, USA, 2Section of Otolaryngology-Head and
Errors in odor identification are associated with sub- Neck Surgery, Department of Surgery, University of Chicago
sequent cognitive impairment in populations clinically Chicago, IL, USA, 3Comparative Human Development
at risk for Alzheimer and Parkinson diseases. However, and Institute for Mind and Biology, University of Chicago
such associations have not been investigated in the gen- Chicago, IL, USA
eral U.S. population. We administered a 5-item test of
odor identification to a U.S. national probability sample In a cross-sectional study we previously demonstrated
of 3,005 community-dwelling adults aged 57–85 in 2005–6 striking race and gender differences in olfactory function
(Wave 1); respondents and their spouses were then retested among older US adults (AChems 2012). Here we report a
in 2010–1 (Wave 2). Odors were presented using Sniffin’ 5 year follow-up of the same subjects to determine 1) the
Sticks, and respondents were asked to identify each from rate of olfactory decline and 2) if this decline differed
among four word-picture options. Respondents in Wave 2 by race or gender. The National Social Life, Health
also completed the Chicago Cognitive Function Measure and Aging Project (NSHAP) interviewed a nationally-
(C-CFM), a survey instrument assessing eight distinct representative sample of community-dwelling older US
cognitive domains, derived from the Montreal Cognitive adults (57–85 years) in 2005–6 (Wave 1) and in 2010–11
Assessment (MoCA). The C-CFM is 30% faster, with (Wave 2), assessing demographics, social life, and health,
scores ranging from 0–20 (mean = 13.5, SD = 4.1) that including olfaction (N = 1436). Odor identification was
correlate highly with the MoCA (r = 0.97). In a multiple measured with 5 Sniffin’ Sticks (0–5 correct). Multivariate
regression model of C-CFM on odor identification score linear regression quantified the association between
in Wave 2 (n = 2,076), each additional odor identification olfactory decline over 5 years and demographic, health
error was associated with a reduction in C-CFM score of and psychosocial factors. Odor identification declined
0.61 (95% CI = (0.44, 0.77); p <0.001), even after control- most rapidly in older individuals (0.25 greater decline in
ling for age, gender, race/ethnicity, education and self-rated number correct per decade of age, P<0.001) and in men
physical and mental health. The change in odor identifi- of all ages (0.32 fewer correct after 5 years vs. only 0.15
cation score over 5 years was also predictive of C-CFM fewer correct for women; P=0.005). Blacks and Whites
scores in Wave 2, with each additional error (relative to declined at the same rate (P=0.925), sustaining the
Wave 1) corresponding to a reduction in C-CFM score of marked cross-sectional race difference observed in Wave
0.29 (95% CI = (0.11, 0.47); p = 0.002). NSHAP is the 1 (equal to 9 years of aging). Neither cognition, SES,
first study to demonstrate an association between olfac- health conditions, mental health, alcohol nor smoking
tory and cognitive function among the U.S. population of predicted accelerated olfactory decline (all P’s>0.05). In
older adults. These results suggest that this relationship addition to having poor olfactory function, men and older
is not limited to those at high risk of neurodegenerative people experience accelerated olfactory decline, effects
disease, but may also represent a more fundamental char- not explained by our measured psychosocial or health
acteristic of the aging process. Acknowledgements: The conditions. We find no evidence of accelerated olfactory
National Social Life, Health and Aging Project Wave 2 aging explaining the health disparity in Blacks seen
(R37 AG030481). DWK is supported by The Center on at the Wave 1 and 2 time points, suggesting that major
Aging Specialized Training Program in the Demography insults to the olfactory system occur before middle age.
and Economics of Aging, which is funded by the National Acknowledgements: The National Institute on Aging
Institute on Aging (NIA) (T32000243). JMP was sup- (R37 AG030481 AG036762 AG029795), the Institute
ported by the Institute of Translational Medicine at The for Translational Medicine at The University of Chicago
University of Chicago (KL2RR025000) and the National (KL2RR025000), the McHugh Otolaryngology Research
Institute on Aging (AG036762). Fund, and the American Geriatrics Society.
#P148 POSTER SESSION III: TRIGEMINAL; in mice while flowing solutions over the anterior or poste-
HUMAN OLFACTORY PSYCHOPHYSICS; TASTE rior tongue separately. Subjects were C57BL/6J mice, which
PERIPHERY were anesthetized prior to surgery to expose the CT. A sili-
cone rubber ring was used to divide the tongue into anterior
Receptor Representations of Perceptual Similarity and posterior halves, and tastants were flowed selectively
Andrew H. Moberly1,2, Lindsey L. Snyder2, Joel D. Mainland1,2 over one half at a time while measuring whole-nerve CT
activity. Responses to NaCl and sucrose were significantly
1
University of Pennsylvania Philadelphia, PA, USA, 2Monell
larger, but responses to HCl and quinine were smaller, when
Chemical Senses Center Philadelphia, PA, USA
stimulating the anterior tongue relative to the posterior. The
In the current consensus model of the olfactory code, the responses to NaCl mixed with amiloride, however, were sim-
recognition of an odorant molecule depends on which recep- ilar for the two regions due to a greater suppressive effect of
tors are activated and to what extent. Here we set out to amiloride on the anterior as compared to posterior tongue.
determine if the receptor-activation profile for a set of cam- Our data suggest that the properties of CT fibers vary
phoraceous odorants is more representative of the structural depending on the location of the taste buds that they inner-
features or the perceptual similarity. The camphoraceous vate. The characteristics of our posterior tongue responses,
odor class is unusual in that despite their common perceptual in fact, were more similar to those normally associated with
odor character the odorants do not share a common func- the glossopharyngeal nerve than with the CT. Additional
tional group (median Tanimoto similarity = 0.14). Using a work will be needed to delineate the relative contributions
set of odorants described as having a camphoraceous qual- of fungiform and foliate papillae to our posterior tongue
ity (Amoore, 1970), we tested the similarity of the odorants responses.
in terms of molecular structure, receptor activation pro-
file in a heterologous assay, and human perceptual ratings.
Molecular structure was quantified using physicochemical #P150 POSTER SESSION III: TRIGEMINAL;
descriptors (Haddad et al., 2008). To measure receptor- HUMAN OLFACTORY PSYCHOPHYSICS; TASTE
activation profiles, we cloned receptors representing 384 of PERIPHERY
the most frequent variants in the 1000 genomes data set. We
then tested the odorants against these receptors using a het- Dried-bonito dashi: Taste qualities evaluated using CTA
erologous luciferase assay. Human perceptual similarity was methods in wild type and T1R1 KO mice.
assayed using an air-dilution olfactometer to obtain pairwise Eugene R. Delay1, Takashi Kondoh2
similarity ratings. Preliminary results suggest that the recep-
tor array is more representative of structural features than of
1
University of Vermont/Department of Biology Burlington,
perceptual similarity. Acknowledgements: R03-DC011373 VT, USA, 2Ajinomoto/Human Health and Nutrition Research
Group Kawasaki, Japan
Dried-bonito dashi, a broth used to increase the
#P149 POSTER SESSION III: TRIGEMINAL; palatability of Japanese cuisine, is made from dried kelp,
HUMAN OLFACTORY PSYCHOPHYSICS; TASTE fish oils, shiitake mushrooms, and other sources. Flavor
PERIPHERY enhancement is due to olfactory and taste stimuli such
as lactic acid, L-amino acids (including glutamate), and
Taste-evoked Chorda Tympani Responses in C57BL/6J inositol monophosphate (IMP). Kawasaki et al. (2008) and
Mice Vary Depending on Which Region of the Tongue is Kondoh et al. (2012) report that rats and mice prefer dashi
Stimulated in 2-bottle preference tests. We conducted conditioned
Rachel M. Dana, Stuart A. McCaughey taste aversion (CTA) experiments to determine what taste
qualities mice can identify in dashi and if an L-amino acid
Ball State University Muncie, IN, USA
receptor (T1R1) contributes to these sensations. C57BL/6J
It is known that each taste bud is responsive to stimuli rep- wild type (WT) mice and T1R1 knockout (KO) mice made
resenting all of the basic taste qualities. However, there is against a mixed background and backcrossed to C57BL
also evidence that the transduction mechanisms that medi- mice (Zhao et al., 2003) were used to see if a CTA to dashi
ate such responses vary depending on location in the oral generalized to (1) 5 basic tastes, and (2) individual L-amino
cavity, and the gustatory nerves differ in amiloride-sensi- acids (+IMP, pH 5.7–5.8) found in dashi (Ajinomoto,
tivity and other properties. Although the peripheral taste- Japan), with or without lactic acid added. The role of odor
responsive nerves have often been compared with each cues was minimized by constant exposure to the odor of
other, there have been few attempts to consider whether the dashi throughout the experiment and by compromising
properties of a particular nerve vary depending on which the nasal epithelium with ZnSO4 (verified by a “chip”
oral regions are stimulated with taste solutions. We there- odor test). Generalization of the CTA was greatest to
fore measured taste-evoked chorda tympani (CT) responses sucrose and weakest to NaCl in WT mice. WT mice also
generalized this CTA to individual amino acids to a degree #P152 POSTER SESSION III: TRIGEMINAL;
roughly related to the concentration of the amino acid in HUMAN OLFACTORY PSYCHOPHYSICS; TASTE
dashi. Lactic acid (1%) altered generalization of the CTA to PERIPHERY
a subset of amino acids. KO mice showed a similar pattern
of generalization to basic tastes but none to L-glutamate. Expression and Signaling of IL-10 in Taste Cells
Even though KO mice readily learned a CTA for dashi, Pu Feng, Jinghua Chai, Hong Wang
they showed minimum generalization to the other L-amino
acids. These results show that dashi elicits a complex taste
and that the T1R1 receptor contributes significantly but Aged taste cells in taste buds are continuously replaced by
not entirely to that complexity. Acknowledgements: NSF young cells differentiated from basal cells. However, the related
IOS-0951016 mechanisms that regulate survival and death of taste cells
remain largely unknown. Here we continue to study the roles
#P151 POSTER SESSION III: TRIGEMINAL; of cytokines tumor necrosis factor (TNF) and interleukin-10
HUMAN OLFACTORY PSYCHOPHYSICS; TASTE (IL-10), in taste cell turnover. TNF, a proinflammatory
PERIPHERY cytokine, often induces apoptosis by binding to its receptors
and activating cell death pathways. In contrast, IL-10, an
GABA as afferent taste neurotransmitter anti-inflammatory cytokine, suppresses TNF expression
Gennady Dvoryanchikov1, Elizabeth Pereira1, Christopher and promotes cell growth and survival. We previously found
Williams2, Stephen D. Roper 1,3, Nirupa Chaudhari 1,3 that T1R3+ type II taste cells are TNF-producing cells, and
TNF receptors are globally expressed in taste buds. Here we
1
Department of Physiology and Biophysics, University report the expression and signaling of IL-10 in taste buds.
of Miami Miller School of Medicine Miami, FL, USA, We find that IL-10 and its receptor IL10R1 are preferentially
2
Graduate Program in Biomedical Sciences, University of expressed by different subsets of type II taste cells. Based
Miami Miller School of Medicine Miami, FL, USA, 3Program on immuno-colocalization experiments using taste cell-type
in Neurosciences, University of Miami Miller School of markers, the IL-10-producing cells are predominantly type
Medicine Miami, FL, USA II taste cells expressing the G-protein α-gustducin, while
ATP is recognized as a principal taste transmitter, acti- IL10R1 is selectively present in taste cells that express T1R3
vating P2X2 and P2X3 purinoceptors located on afferent and TNF. The IL-10 production can be induced by microbial
sensory fibers that innervate taste buds. Taste buds also products lipopolysaccharide (LPS) and Staphylococcal
contain and release several other neurotransmitters. Many Enterotoxin A (SEA). The LPS-induced IL-10 expression in
of these have been shown to mediate cell-to-cell communi- taste cells was profoundly diminished in TLR2-TLR4 double-
cation within the bud. For instance, we recently reported gene-knockout mice, which indicates the dependence of IL-10
that GABA serves as an inhibitory transmitter, decreas- induction on TLR signaling pathways. The results strongly
ing taste-evoked ATP release from taste buds. We now ask suggest that IL-10 produced by α-gustducin+ type II cells
whether GABA might also act on afferent sensory axons could specifically down-regulate TNF production by acting on
that innervate taste buds. To answer this question, we ana- IL10R in T1R3+TNF+ type II cells. This interaction between
lyzed GABA receptors in geniculate ganglia. Neurons in the two subsets of taste cells may provide a mechanism for
this ganglion innervate fungiform and palatal taste buds. cellular crosstalk in taste buds under circumstances such as
Using RT-PCR, we show that GABA-A α 1 is prominently injury, infection and inflammation. Acknowledgements: The
expressed along with lower levels of α2, β2, β3, γ1 and study was supported by NIH/NIDCD grants DC010012 and
γ2 subunits. The cation-chloride cotransporters, KCC3 DC011735 and NSF grant DBJ-0216310
and KCC4, and particularly, NKCC1, which is essential
for the inhibitory action of GABA, are also expressed in
geniculate ganglia. Immunostaining shows that a majority #P153 POSTER SESSION III: TRIGEMINAL;
of geniculate ganglion neurons express variable levels of HUMAN OLFACTORY PSYCHOPHYSICS; TASTE
GABAA-α1 in their somata. Fibers projecting peripher- PERIPHERY
ally from geniculate ganglia were strongly immunopositive
for GABA-α1, a pattern very similar between GABAA-α1 Bitter Taste Stimuli trigger functionally distinct and
and P2X2. In summary, GABA may function for both cell- interdependent Gustatory Signaling Pathways in
to-cell communications within taste buds and in afferent immortalized Human Taste Cells
taste neurotransmission. Functional imaging studies are in Andreas Hochheimer, Michael Krohn, Holger Zinke
progress to test whether inhibitory responses to GABA are
B.R.A.I.N AG Zwingenberg, Germany
detected in all or a subset of geniculate ganglion neurons.
Acknowledgements: NIH/NIDCD R01DC6308 and NIH/ Stably proliferating human taste cell lines are a powerful tool
NIDCD R21DC12746 to gain new insights into human taste reception and signal
transduction mechanisms. We previously established human and the identity of the cells that respond to Na+ stimulation
taste cell lines from lingual epithelium, which maintain their remain controversial. Using Ca2+ imaging on isolated mouse
endogenous programming and dedication to bitter taste vallate taste cells loaded with Fura 2, we demonstrate here that
reception. HTC-8 cells express 15 of 25 human TAS2R bit- a subset of cells, neither Receptor nor Presynaptic cells, show
ter taste receptor genes and respond to various bitter stimuli Ca2+ mobilization in response to Na+ stimulation. Removing
with endogenous gustatory signaling including calcium sign- extracellular Ca2+ had no effect on responses evoked by Na+
aling and cell membrane depolarization. We used Fluo-4 stimulation. In marked contrast, applying 1 µM thapsigargin,
calcium imaging assays and FLIPR fluorescent membrane a SER Ca-ATPase inhibitor, or 10 µM U73122, a PLC blocker,
potential assays to measure responses of human taste cells to eliminated Na+ responses, suggesting that Na+ stimulation
various bitter taste stimuli and combinations of bitter taste triggers PLC/IP3-mediated release of Ca2+ from intracellular
stimuli. Our results revealed that stimulation with salicin elic- stores. Next, we used CHO cells expressing P2X receptors
its a PLC-dependent increase of intracellular calcium from as biosensors to monitor ATP release from taste buds and
internal calcium stores and does not lead to membrane depo- isolated taste cells. Na+ stimulation elicited robust biosensor
larization. In contrast, addition of other bitter taste stimuli responses that were blocked by suramin, confirming that
for instance PTC, saccharin and aristolochic acid led to cell Na+ stimulation elicits ATP secretion from taste buds/cells.
membrane depolarization as well as to PLC-independent Carbenoxolone (5 µM) or probenecid (250 µM), blockers
increase of intracellular calcium, which depends on extracel- of pannexin1 hemichannels, reversibly blocked Na+- evoked
lular calcium. These results suggest that gustatory responses ATP secretion. Our data indicate that salt taste stimulus
to bitter stimuli are not uniform in human taste cells and that triggers a dedicated population of taste cells to secrete
bitter taste stimuli may trigger distinct signaling pathways. ATP via pannexin1hemmichannels. Acknowledgements:
To test, whether these distinct signaling pathways interact we Supported by NIH/NIDCD 5R01DC007630 (SDR).
stimulated HTC-8 cells with salicin in combination with PTC,
saccharin or aristolochic acid in the absence of extracellular
calcium. Surprisingly, even though PTC, sacharin and aris- #P155 POSTER SESSION III: TRIGEMINAL;
tolochic acid alone elicited no response, the PLC-dependent HUMAN OLFACTORY PSYCHOPHYSICS; TASTE
increase of intracellular calcium in response to salicin was PERIPHERY
strongly enhanced. These results suggest that crosstalk
between bitter taste receptors and/or signaling pathways may IL-1β Enhances Sodium Transport in Taste Buds
occur in human taste cells. Acknowledgements: The research D Kumarhia
was funded by BRAIN AG corporate funds.
Institute of Molecular Medicine and Genetics Augusta,
GA, USA
#P154 POSTER SESSION III: TRIGEMINAL; Sodium ions pass through apical epithelial sodium channels
HUMAN OLFACTORY PSYCHOPHYSICS; TASTE (ENaC) in taste cells, depolarizing them and transmitting
PERIPHERY information about sodium taste to the brain. Salt taste trans-
duction is among the least understood of the taste transduc-
Sodium-sensing cells in mouse vallate taste buds tion pathways. Moreover, few ENaC modulators have been
Anthony Huang1,3, Stephen D Roper1,2 identified in taste receptor cells compared to epithelial cells
from kidney, lung and colon. Interleukin (IL)-1β is a classi-
Department of Physiology & Biophysics, University of
1
cal proinflammatory cytokine produced by activated leuko-
MiamiMiller School of Medicine Miami, FL, USA, 2Program in
cytes. IL-1β and its receptor are also strongly expressed in
Neuroscience, University of Miami Miller School of Medicine
taste cells. This cytokine promotes the maintenance of nor-
Miami, FL, USA, 3Department of Anatomy, Southern Illinois
mal sodium taste function after contralateral chorda tympani
University Carbondale, IL, USA
injury, but the direct effects of IL-1β on sodium transport in
Taste buds contain two types of cells that directly taste buds are unknown. We loaded rat lingual epithelia con-
participate in gustatory transduction-- Receptor (Type II) taining fungiform taste buds with the sodium indicator dye
and Presynaptic (Type III) cells. Receptor cells respond to CoroNa Green, and measured changes in fluorescence (F490)
sweet, bitter and umami taste stimulation. Presynaptic cells in response to basolateral IL-1β. Apical administration of the
have been identified as sour (acid) taste-sensing cells. Using ENaC blocker, amiloride (50 μM in control Ringer’s solution),
confocal Ca2+ imaging in a lingual slice preparation, Tomchik decreased F490 by 10–15%. Basolateral IL-1β (0.05 ng/ml–5 ng/
et al. (2007) showed that some vallate taste cells responded to ml in control Ringer’s solution) caused upsurges in F490 result-
salt (Na+) taste. Recently, ion channels (ENaCs) believed to ing in an overall 2–10% increase in sodium transport above
underlie salt taste transduction were reported to be in taste baseline. The effects of IL-1β were at least partially amiloride-
cells that were neither Receptor (Type II) nor Presynaptic sensitive, and occurred within seconds. These results indicate
(Type III) cells. Yet, mechanisms of Na+ taste transduction that IL-1β rapidly augments ENaC function, in contrast to
the cytokine’s delayed inhibition of sodium transport in other University at Buffalo Buffalo, NY, USA, 2John B Pierce Lab
1
epithelial cells. Together, this indicates a novel, direct influence New Haven, CT, USA
of IL-1β on ENaC in taste buds, mediated by mechanisms We reported that ryanodine receptors, specifically ryanodine
which diverge from those acting in other sodium-transporting receptor type 1, are expressed in two different types of mam-
epithelia. Acknowledgements: NIDCD 005811-10 malian peripheral taste receptor cells: Type II and Type III
cells. In Type II cells that lack voltage-gated calcium chan-
#P156 POSTER SESSION III: TRIGEMINAL; nels (VGCCs) and chemical synapses, the ryanodine recep-
HUMAN OLFACTORY PSYCHOPHYSICS; TASTE tors contributed to the taste-evoked calcium signals that are
PERIPHERY initiated by opening inositol trisphosphate receptors located
Ligand Specificity of Orphan G Protein-coupled on internal calcium stores. In Type III cells that do have
Receptor GPR84 VGCCs and chemical synapses, ryanodine rreceptors were
no longer able to contribute to taste-evoked calcium release
Yan Liu, Timothy A. Gilbertson signals but contributed to the depolarization-dependent
Department of Biology, Utah State University Logan, calcium influx. The goal of this study was to better under-
UT, USA stand the role of the ryanodine rreceptors in Type III cells.
Specifically, we wished to establish if there was selectivity in
Previous studies concluded that medium chain fatty acids
the type of VGCC that was associated with the ryanodine
with carbon chain lengths of 9–14 were ligands for GPR84
receptor or if the ryanodine receptor opened irrespective of
(Venkataraman and Kuo Immunol Lett 101:144, 2005; Wang
the calcium channels involved. We also wished to determine
et al. J Biol Chem 281:34457, 2006), however, there has
if the ryanodine receptors and VGCCs required a physical
never been a careful and systematic analysis of the ligand
linkage to interact or were simply functionally associated
specificity of this receptor which we have shown previously
with each other. Using calcium imaging and pharmacologi-
to be expressed in mammalian taste cells. As a means to
cal inhibitors on a transgenic mouse line that expresses green
compare the specificity and concentration-response functions
fluorescent protein (GFP) in GAD67 expressing Type III
for fatty acids in the taste system with GPR84, fura-2 based
taste cells, we found that ryanodine receptors are selectively
ratiometric calcium imaging was used to characterize GPR84
associated with L type VGCCs but not through a physical
in a cell line that has been designed to express this receptor in
linkage. Taste cells are able to undergo calcium induced cal-
an inducible fashion under control by the tetracycline (TET)
cium release through ryanodine receptors to increase the
promoter. The specific cell line has an inducible GPR84
initial calcium influx signal and provide a larger calcium
+ Gqi9 (a chimeric G protein; (Wang et al., 2006)), which
response than would otherwise occur when L type channels
has been cloned, validated by PCR/qPCR and verified for
are activated. Acknowledgements: This work was supported
function in FLIPR-based calcium assays. Non- induced cells
by NSF Grant 0917893 to KFM.
were used as controls. Using fura-2 based calcium imaging, we
found that caproic (C6:0), caprylic acid (C8:0), capric acid (C10:0),
undecanoic acid (C11:0), lauric acid (C12:0), oleic acid (C18:1), and #P158 POSTER SESSION III: TRIGEMINAL;
arachidic acid (C20:0) can all elicited a robust and reversible HUMAN OLFACTORY PSYCHOPHYSICS; TASTE
increase in intracellular calcium in the cells induced to express PERIPHERY
GPR84, while they cannot induce any calcium signal in the
non-induced cells. Our results suggest GPR84 functions as a Functional Profile of the Adult Glossopharyngeal Nerve
receptor for unsaturated fatty acids from C6:0–C12:0 and other Following Neonatal Chorda Tympani Transection in Rats
fatty acids (oleic acid, arachidic acid) previously not thought Louis J. Martin, Suzanne I. Sollars
to activate this receptor. Currently, we are investigating the
concentration-response functions for ligands of GPR84 in University of Nebraska at Omaha/Psychology Department
the inducible cell line. The specific signaling pathway for fatty Omaha, NE, USA
acid transduction through GPR84 receptors and its role in the Rats receiving bilateral neonatal chorda tympani transection
peripheral gustatory system remain to be elucidated. (neoCTX) show an increased preference for ammonium
chloride (NH4 Cl) as adults – a substance which normal adult
#P157 POSTER SESSION III: TRIGEMINAL; rats never prefer. It is currently unclear what changes to the
HUMAN OLFACTORY PSYCHOPHYSICS; TASTE taste system underlie this altered preference. To determine
PERIPHERY if injury-induced differences in the response properties of
the remaining taste nerves can account for this behavior,
Ryanodine receptors selectively interact with L type calcium
whole-nerve electrophysiology was performed on the
channels in mouse taste cells
glossopharyngeal nerve (GL) of adult rats that underwent
Amanda B Maliphol1, Michelle R Rebello1,2, Kathryn F either unilateral neoCTX or a control surgery at five days
Medler1 of age. Nerve activity following application of various
concentrations of NH4Cl, NaCl, and sucrose was recorded entire population of bitter sensing cells but only ~50%. The
using 0.5M NH4Cl and 0.5M KCl as standards. There were remaining bitter sensing cells displayed complete absence of
no differences in nerve response to NH4Cl or NaCl, but there some Tas2rs, whereas expression of others was reduced to
was a significant difference for sucrose, with neoCTX rats different extents. Behavioral experiments showed that these
having higher GL responses to the tastant. Since NH4Cl mice exhibit diminished avoidance of several bitter com-
responses did not differ between surgical groups, there pounds, whereas they are indistinguishable from control
may be differences following neoCTX in greater superficial mice in their avoidance of denatonium benzoate. Together
petrosal or superior laryngeal nerve activity, or alterations the data demonstrate that bitter sensing cells are function-
in central processing which can account for the increase ally polymorphic forming the basis for variable behavioral
in NH4Cl preference. Alternatively, the functioning of the responses to different bitter chemicals.
GL may be more affected following bilateral compared
to unilateral neoCTX. The mechanisms which lead to the
observed injury-induced alteration in sucrose responding are #P160 POSTER SESSION III: TRIGEMINAL;
unknown. However, this increase in responding following HUMAN OLFACTORY PSYCHOPHYSICS; TASTE
neoCTX could help explain observations from the human PERIPHERY
literature in which early chorda tympani damage is correlated Behavioral responses to trimethylamine -N -oxide using the
with an increased preference for sugary foods. Work is CTA paradigm in C57BL/6 mice
currently underway to record GL responses from adult rats
receiving bilateral neoCTX at 10 days of age. Yuko Murata, Meiko Kimura
Fisheries Research Agency Yokohama, Japan
Trimethylamine-N-oxide (TMAO) is a common and com-
#P159 POSTER SESSION III: TRIGEMINAL; patible osmolyte in tissues of marine organisms, and coun-
HUMAN OLFACTORY PSYCHOPHYSICS; TASTE teracts the effects of protein destabilizing agents such as
PERIPHERY urea in elasmobranches. Gadoid fish, elasmobranches and
Mouse bitter taste scallop have high levels of TMAO in their muscles. TMAO
is thought to contribute to the taste of these fishes but the
Wolfgang Meyerhof1, Kristina Lossow1, Hübner Sandra1,
taste of TMAO is unclear. In this study, we investigated taste
Frenzel Sabine1, Masataka Narukawa1, Anja Voigt1, Ulrich
quality perception of TMAO in C57BL/6 mice using con-
Boehm2, Jonas Töle1, Maik Behrens1
ditioned taste aversion (CTA) experiments.We developed
1
German Institute of Human Nutrition Potsdam-Rehbruecke, LiCl-induced CTA to 1.0% TMAO and examined its gener-
Department of Molecular Genetics Nuthetal, Germany, alization to 12 taste stimuli. CTA to TMAO significantly gen-
2
University of Saarland, School of Medicine, Department of eralized to D-phenylalanine, saccharine and quinine. These
Pharmacology and Toxicology Homburg, Germany results suggest that mice avoid the taste of TMAO and its
taste perception in mice is similar to D-phenylalanine, sac-
Depending on dose, bitter chemicals can be toxic or healthy.
charin and quinine. We will also present behavioral thresh-
Accordingly, consumers like some bitter tasting foods while
olds and behavioral response to low concentration (below
they avoid others. For a detailed understanding of bitter
0.1%) of TMAO.
taste physiology we examined the receptors for bitter sub-
stances and their cells in mice. Functional expression analy-
sis showed that mice and humans detect a similar range of #P161 POSTER SESSION III: TRIGEMINAL;
bitter compounds even though mice possess 30% more Tas2r HUMAN OLFACTORY PSYCHOPHYSICS; TASTE
bitter taste receptor genes than humans. Intriguingly, recog- PERIPHERY
nition of bitter chemicals in the two species is mostly evoked
by non-orthologous Tas2rs. Based on the number of cognate Purification, biophysical characterization and first
compounds, mouse Tas2rs, like their human counterparts, crystallization trials of the ligand-binding domain of the
can be classified in generalists, moderately tuned receptors human T1R3 sweet taste receptor.
and specialists. However, compared with humans, mice seem Fabrice Neiers, Maud Sigoillot, Elodie Maîtrepierre, Christine
to possess a larger fraction of specialists suggesting that a Belloir, Nicolas Poirier, Loïc Briand
greater number of Tas2r genes offers the luxury of a set of
Centre des Sciences du Goût et de l’Alimentation, INRA
specific receptors for selected bitter compounds. Genetic
CNRS Université de Bourgogne Dijon, France
labeling and in situ hybridization experiments revealed that
mice possess 2200 to 3300 bitter-sensing cells. They express The heterodimeric T1R2/T1R3 sweet taste receptor is composed
the entire repertoire of Tas2r genes at individual levels and of two class C G-protein coupled receptors (GPCRs), while
in limited subsets. Accordingly, genetic ablation of the cells T1R1/T1R3 heterodimer is involved in umami taste perception.
expressing one Tas2r, i.e., Tas2r131, did not extinguish the class C GPCRs share common structural homologies
including a large N-terminal domain (NTD) linked to the Lep may act as a modulator in mice that tonically influence
seven transmembrane domain by a cysteine rich region. T1R2- basal sweet sensitivity, while ECs may become more effective
and T1R1-NTDs have been shown to contain the primary with defects in Lep system. Acknowledgements: Supported
binding site for most of the sweet ligands and umami tastants, by JSPS KAKENHI Grant Number 18077004, 18109013,
respectively. In contrast, the contribution of T1R3-NTD to 23249081 (YN) and 21791808, 23689076 (RY).
sweet and umami compound detection is less documented. The
human T1R3-NTD was produced in Escherichia coli using a
strategy recently described (Maîtrepierre et al., Protein Expr. #P163 POSTER SESSION III: TRIGEMINAL;
Purif., 2011). The purified protein was characterized by SDS– HUMAN OLFACTORY PSYCHOPHYSICS; TASTE
PAGE, circular dichroism and a fluorescent ligand-binding PERIPHERY
assay. Using size exclusion chromatography coupled to static
light scattering, we showed that hT1R3-NTD is monodisperse Transcriptome analysis of individual taste receptor cells
and forms homodimers. These data demonstrate that the Sunil K Sukumaran, Brian C Lewandowski, Yuri Kaulin,
purified protein is not only suitable for functional analyses, but Peihua Jiang, Gary Beauchamp, Alexander Bachmanov,
also for subsequent crystallization trials. Acknowledgements: Liquan Huang, Robert F Margolskee
This work was supported by fundings from INRA, Burgundy
council (Région Bourgogne) and Agence Nationale de la
Recherche (ANR-09- ALIA-010). Although many genes important for taste receptor cell
functioning have been identified, including Tas1r and Tas2r taste
#P162 POSTER SESSION III: TRIGEMINAL; receptors and downstream signaling components a-gustducin
HUMAN OLFACTORY PSYCHOPHYSICS; TASTE and Trpm5, much remains unknown about these cells. In this
study, we have taken a whole genome transcriptomics approach
PERIPHERY
to identify genes selectively expressed in individual mouse taste
Regulation of basal sweet sensitivity of mice by leptin. receptor cells of specific varying subtypes. We standardized
Mayu Niki, Masafumi Jyotaki, Tadahiro Ohkuri, Ryusuke methods for isolating single taste receptor cells and linearly
Yoshida, Yuzo Ninomiya amplifying their mRNA population using a T7-OligodT
primed method. Transcriptome profiling was done by deep
Section of Oral Neuroscience, Graduate School of Dental sequencing using the illumina HiSeq platform and Affymetrix
Sciences, Kyushu University Fukuoka, Japan microarrays. Statistical analysis of each individual taste cell’s
Leptin (Lep) is shown to selectively suppress sweet taste transcriptome data from fifteen Tas1R3 expressing type II
responses in lean mice but not in Lep receptor-deficient db/ cells, seventeen NaCl responsive type III cells, eight NaCl
db mice. In contrast, endocannabinoids (EDs) enhance sweet unresponsive type II cells and five Lgr5 expressing stem/
taste sensitivities in lean mice but not in mice genetically progenitor cells demonstrates that these cell types form distinct
lacking CB receptors. However the action of endogenous groups. Further, many marker genes specific to each particular
Lep and EDs on taste responses is not fully understood. taste cell type are differentially expressed several thousand-fold
In this study, we examined expression of related molecules, to several million- fold, confirming the reliability of our method.
the effect of leptin on taste cell responses and the effect of We are using this method to identify pathways unique to each
antagonists for Ob-Rb (leptin L39A/D40A/F41A: LA) and type of mature taste receptor cell as well as to stem/precursor
CB (AM251) on the chorda tympani (CT) responses in cells. We are using data-mining to discover hitherto unknown
mice with different serum Lep levels. About 40 % of taste functionalities of taste cells. Our method also can be applied
cells expressing T1r3 coexpressed Ob-Rb and a subset of to other cell types as well. Acknowledgements: NIH-NIDCD
taste cells expressed biosynthesizing enzyme (DAGL) and Core Grant 1P30DC011735-02 to Monell, NIH-DC03155
degrading enzyme (MAGL) of ED (2- AG). In about half to RFM and NIH-R01DC010842, Commonwealth of
of sweet sensitive taste cells, bath application of 20 ng/ml Pennsylvania Department of Health to PJ
leptin suppressed responses to sweeteners (<80% of control
response). The effect of leptin was concentration dependent
and reached maximal level at 10–20 ng/ml. When the cells #P164 POSTER SESSION III: TRIGEMINAL;
were adapted to several concentrations (1–5 ng/ml) of lep- HUMAN OLFACTORY PSYCHOPHYSICS; TASTE
tin, increases in 10 ng/ml leptin still affected sweet responses PERIPHERY
of taste cells. Administration of LA significantly increased
The L-Glutamate Chemoreceptor of Paramecium, an
CT responses to sweeteners in lean mice, whereas admin-
Ortholog of NMDA-like Receptor, is located in the Soma
istration of AM251 did not affect. Moreover the effect of
and Ciliary membranes
LA on CT responses to sweeteners gradually decreased with
increasing plasma leptin levels, whereas the reverse is true Judith L. Van Houten1, Mallory P. Romanovitch2, Cassandra
for ECs. These results suggest a possibility that circulating Jacobs3, Megan Valentine1, Junji Yano1
University of Vermont Burlington, VT, USA, 2Transgenomic

1
qualities require ATP to communicate with nerve fibers.
New Haven, CT, USA, 3Duke University Durham, NC, USA However, subsequent studies have detected ATP release
Paramecium tetraurelia is strongly attracted to L-glutamate. only from Type II taste cells, those that respond to bitter,
The cells rapidly hyperpolarize in L-glutamate from a K sweet, and umami stimuli (Huang et al., 2007; Romanov
conductance (Preston and Usherwood, 1988), causing et al., 2007). Recent experiments on the P2X2-P2X3 DKO
smooth and rapid swimming and attraction to the stimulus. mice have shown that in addition to the lack of postsynaptic
Upon binding to its receptor, L-glutamate initiates a rapid receptors, these mice fail to release ATP to taste stimuli
increase in intracellular cyclic AMP (Yang et al., 1997). (Huang et al., 2011). Thus, the lack of taste responses may
Downstream from this second messenger is the activation be due to the lack of ATP release rather than the lack of
of Protein Kinase A, that activates the plasma membrane postsynaptic receptors. To resolve whether postsynaptic P2X
calcium pumps to sustain the hyperpolarizing conductance receptors are required for transmission of all tastes to the
and sustain the smooth fast swimming. We have used RNA nervous system, we have used a pharmacological approach
interference (RNAi) to test several candidate genes for pos- to chemically block the purinergic receptors while recording
sible function as an L-glutamate receptor and found a gene from the chorda tympani nerve in response to a battery
with sequence homology to the glutamate binding subunit of taste stimuli. C57Bl6 mice were injected ip with 6 mg/
of an NMDA-like receptor. When the mRNA for this gene, kg AF-353, a membrane permeant compound that blocks
pGluR1, is depleted using RNAi, the cells are not attracted all P2X3-containing receptors (P2X3 homotrimers and
to L-glutamate, but continue to be attracted to D-glutamate. P2X2/3 heterotrimers; Gever et al., 2010). Within 15 min of
To better understand the signaling pathway for L-glutamate, injection, integrated responses to all taste stimuli, including
we have epitope tagged the pGluR1 protein and found by acids and salts, were abolished. Taste responses also are
Western blotting and mass spectrometry that it is in the mem- blocked in a dose-dependent fashion with application of
brane of the cell body and cilia. A catalytic subunit of the higher concentrations of AF-353 directly to the tongue and
ciliary adenylyl-cyclase labeled with GFP does not co-immu- can be partially recovered upon washout of the drug. These
niprecipitate with FLAG-pGluR1. Mammalian NMDA-like data clearly indicate that activation of P2X receptors and
receptors are dependent upon glycine and D-serine, but the therefore ATP release is required for all taste modalities
P. tetraurelia chemoresponse to L-glutamate is unaffected in mice. Acknowledgements: Funded by NIH grants R01
by combinations of glycine and D-serine and L-glutamate. DC012555, R01 DC007495, and P30 DC04657 and a gift
Acknowledgements: NIH Grant Number 2 P20 RR016435- from Afferent Pharmaceuticals.
06 to the Neuroscience Center of Research Excellence
(COBRE to R. Parsons, PI) for imaging support and NIH
grant RO1 GM59988 #P166 POSTER SESSION III: TRIGEMINAL;
PERIPHERY
Nonsynaptic Contacts in Rat Circumvallate Taste Buds:
Subsurface Cisternae and Atypical Mitochondria
PERIPHERY
Ruibiao Yang1,2, Amanda E. Bond1,2, John C. Kinnamon1,2
A selective P2X3, P2X2/3 receptor antagonist abolishes
responses to all taste stimuli in mice Department of Biological Sciences, University of Denver
1
Denver, CO, USA, 2Rocky Mountain Taste and Smell Center

Aurelie Vandenbeuch1,3, Catherine B. Anderson1,3,
Aurora, CO, USA
Anthony P. Ford4, Steve Smith4, Thomas E. Finger2,3, Sue
C. Kinnamon1,3 It is generally accepted that Type II cells transduce bitter,
sweet, and umami stimuli in rodent taste buds. Recent stud-
1
University of Colorado School of Medecine, Department of
ies also demonstrate that Type II taste cells release ATP, a
Otolaryngology Aurora, CO, USA, 2University of Colorado
neurotransmitter believed to play an important role in taste
School of medecine, Department of Cell and Development
transduction. However, no classical synapses have been
Biology Aurora, CO, USA, 3Rocky Mountain Taste and Smell
found to be associated with Type II cells. Are there other
Center Aurora, CO, USA, 4Afferent Pharmaceuticals San
contacts between Type II cells and Type III cells or Type II
Mateo, CA, USA
cells and nerve processes? In the present study, we utilized
ATP is believed to be a crucial neurotransmitter for conventional transmission electron microscopy to examine
communicating gustatory information to nerve fibers. the ultastructural features of nonsynaptic contacts in rat
Evidence is based largely on recordings from mice lacking circumvallate taste buds. Our results indicate that Type II
both P2X2 and P2X3 purinergic receptor subunits (P2X2- cells are in intimate contact with Type III cells in taste buds.
P2X3 DKO mice), which lack responses to all taste stimuli Two types of nonsynaptic contacts, subsurface cisternae of
(Finger et al., 2005). These data suggest that all taste endoplasmic reticulum and/or atypical mitochondria, have
been found to be present adjacent to the cytoplasmic leaf- (T4). Consistent with this, we found that a nonsynonymous
let of Type II cells at close appositions with nerve processes. polymorphism in T2R42 is associated with lower free T3
Frequently we observed subsurface cisternae of smooth or and T4 levels in a human cohort. Thyroid hormones can
rough endoplasmic reticulum in Type II cells at sites of appo- affect energy expenditure, thermoregulation, body weight,
sition with nerve processes. Occasionally we observed atypi- and body composition. T2Rs may be a useful target for
cal mitochondria and subsurface cisternae adjacent to each pharmacologic regulation of these endocrine signals,
other in Type II cells. We also observed electron-dense peri- perhaps leading to new interventions for chronic morbidities
odic pillars connecting the subsurface cisternae or the atypi- involving fatigue or obesity. Acknowledgements: Support:
cal mitochondria to the membranes of Type II cells at close NIDCD (R01 DC010110), NIDDK (P30 DK072488).
appositions with nerve processes. We speculate that these
structures are involved in communication between Type II
cells and nerve processes. Acknowledgements: This work is #P168 POSTER SESSION IV: CHEMICAL
supported by NIH grants DC00285 and P30 DC04657 SIGNALING AND BEHAVIOR; ANIMAL
BEHAVIOR/PSYCHOPHYSICS;
CHEMOSENSATION AND METABOLISM;
#P167 POSTER SESSION IV: CHEMICAL VOMERONSASAL AND CHEMICAL
SIGNALING AND BEHAVIOR; ANIMAL COMMUNICATION
Metabolic effects of long-term sweetener consumption
VOMERONSASAL AND CHEMICAL Maartje CP Geraedts1, Tatsuyuki Takahashi1, Stephan
COMMUNICATION Vigues1, Cedric Uytingco1, Steven D Munger1,2
A role for bitter taste receptors in thyroid toxicity and

1
University of Maryland School of Medicine, Department of
hormone production Anatomy & Neurobiology Baltimore, MD, USA, 2University
of Maryland School of Medicine, Department of Medicine,
Adam A. Clark1,2, Cedrick D. Dotson1, Amanda E.T. Elson1,
Division of Endocrinology, Diabetes and Nutrition Baltimore,
Nanette I. Steinle3, Steven D. Munger1,2,3
MD, USA
1
University of Maryland Baltimore Department of Anatomy
The sweet taste receptor T1R2+T1R3 responds to diverse
and Neurobiology Baltimore, MD, USA, 2University of
gustatory stimuli including sugars and low calorie sweeten-
Maryland Baltimore Program in Toxicology Baltimore, MD,
ers (LCS). In intestinal enteroendocrine L cells, T1R2+T1R3
USA, 3University of Maryland Baltimore Department of
couples glucose stimulation to the secretion of insulinotropic
Medicine Baltimore, MD, USA
hormone glucagon-like peptide-1 (GLP-1). While the prepon-
Bitterness is associated with toxicity. Bitter compounds derance of in vivo evidence indicates that LCS do not impact
activate a small family of G protein-coupled taste glucose homeostasis, the responsiveness of T1R2+T1R3 to
receptors (T2Rs) expressed in the taste buds. T2Rs are also natural and artificial LCS suggests that these compounds
found in non-gustatory tissues (e.g., the respiratory and could exert extraoral effects. To address this issue, we have ini-
gastrointestinal systems), suggesting that many ingested or tiated long-term sweetener (300 mM sucrose, Su; 1 mM sucra-
inhaled toxins could have broad physiologic effects. We report lose, Sa; 10 mM cyclamate, Cy; 3 mM acesulfame K; 5 mM
that T2Rs are expressed in thyroid follicular cells (FCs), rebaudioside A) consumption studies in mice with (T1R3+/+)
where they modulate iodide efflux. Immunohistochemical or without (T1R3−/−) a functional sweet taste receptor. Mice
and PCR analyses show that numerous T2Rs as well as the (4 w.o.) are maintained for 3, 6, 9 or 12 mo. on normal chow
T2R-associated G protein subunit α-gustducin are expressed diets along with ad lib water containing one of the sweeten-
in human and rodent FCs and in a human follicular cell ers. Cy serves as a negative control as it is not an agonist for
line, Nthy-Ori 3-1. Thyroid stimulating hormone (TSH)- mouse T1R2+T1R3. Mice are assessed for food and fluid
dependent Ca2+ signals, an important regulator of iodide intake, sweet taste preference, body weight, body fat, met-
efflux from FCs, were significantly reduced in Nthy- Ori rics of glucose and insulin homeostasis, and glucose-stimu-
3-1 cells by the T2R ligands denatonium benzoate (DB), lated GLP-1 secretion from ileum and colon explants. Initial
chloramphenicol (Chlor) and cycloheximide (Cyx). By results indicate that both T1R3+/+ and −/− mice maintained on
contrast, the T2R38 ligand 6-n-propylthiouracil (PROP) 300mM Su have significantly higher body fat content than
had no effect on TSH-dependent Ca2+ signals, likely either Sa- or Cy-treated mice. By contrast, Sa-treated mice
because this cell line expresses only the “non-taster” T2R38 show a strong potentiation of glucose-stimulated GLP-1
variant. DB, Chlor and Cyx also significantly reduced TSH- secretion (as compared to Su- or Cy-treated mice) at 3 mo,
dependent iodide efflux from Nthy-Ori 3-1 cells. Decreased with reduced potentiation at later time points. Interestingly,
iodide efflux in vivo should result in decreased production of glucose tolerance tests revealed no significant differences
the thyroid hormones triiodothyronine (T3) and thyroxine in glucose or insulin levels across these sweetener groups.
Ongoing assessments of all five sweeteners will be presented. #P170 POSTER SESSION IV: CHEMICAL
Acknowledgements: Tate&Lyle and NIDCD (DC010110) SIGNALING AND BEHAVIOR; ANIMAL
#P169 POSTER SESSION IV: CHEMICAL
VOMERONSASAL AND CHEMICAL
SIGNALING AND BEHAVIOR; ANIMAL
COMMUNICATION
CHEMOSENSATION AND METABOLISM; As American as Apple Pie Gum: A Study of Satiety
VOMERONSASAL AND CHEMICAL Jack Hirsch1, Michele Soto2, Alan Hirsch2
COMMUNICATION
Adlai E. Stevenson High School Lincolnshire, IL, USA, 2Smell
1
Oral stimulation with sugar elicits cephalic phase insulin & Taste Treatment and Research Foundation Chicago, IL,
release and improves glucose tolerance in C57BL/6 and USA
Tas1r3 knock-out mice
Purpose: The aim of this study is to determine the satiety
John I. Glendinning1, Sarah Stano1, Olivia Goldman1, Danica value of Wrigley’s Extra Dessert Delight Apple Pie Sugar-
Yang1, Robert F. Margolskee2, Anthony Sclafani3, Joseph Free Gum as compared to a 100 calorie slice of apple pie.
R. Vasselli4 Procedure: Thirteen girls and 11 boys self-assessed their
1
Barnard College, Columbia University/Biology Department degree of satiety on a visual analog scale before and after
New York, NY, USA, 2Monell Chemical Senses Center chewing Wrigley’s Extra Dessert Delight Apple Pie Sugar-
Philadelphia, PA, USA, 3Brooklyn College of CUNY/ Free Gum for one, 15, and 30 minutes. This was repeated
Psychology Department Brooklyn, NY, USA, 4New York on a separate day with a 100 calorie slice of Market Pantry
Obesity Nutrition Research Center, St. Luke’s- Roosevelt Apple Pie (Target) or vice-versa (the order of presentation
Hospital New York, NY, USA being counter balanced). A satiety value was computed for
each and statistical significance was determined for differ-
In rats, oral stimulation by sugars elicits cephalic phase ence in satiety index between the two and effect of order
insulin release (CPIR). This CPIR has been found to improve of presentation (pie versus gum first). Conclusion: No sta-
tolerance to glucose challenges. Here, we asked whether oral tistically significant difference was seen in satiety value in
stimulation by glucose has similar effects in C57BL/6 wild- response to eating a slice of apple pie or chewing apple pie-
type (WT) mice. To explore the role of the T1r3 subunit of flavored gum (p=0.096). No effect of order presentation was
the heterodimeric sweet taste receptor (T1r2+T1r3) in this seen. The gum imbued the same satiety value as apple pie,
process, we also examined Tas1r3 knock-out (KO) mice. with less than 1/20 of the calories. The results suggest that
We subjected all mice to a 2 g/kg glucose challenge. To chewing Wrigley’s Extra Dessert Delight Apple Pie Sugar-
this end, we administered a 2.8 M glucose solution either Free Gum may have a role in the promotion of satiety in
orally (by allowing mice to take a predetermined number children as part of a weight loss program.
of licks) or post-orally (by injecting a specific volume into
the esophagus by feeding tube). We collected tail blood
immediately prior to glucose administration (at 0 min), and #P171 POSTER SESSION IV: CHEMICAL
at subsequent time intervals. In Experiment 1, we measured SIGNALING AND BEHAVIOR; ANIMAL
plasma insulin levels with an ELISA test. We inferred a BEHAVIOR/PSYCHOPHYSICS;
CPIR if plasma insulin increased significantly between the CHEMOSENSATION AND METABOLISM;
0 and 5 min measurements. Both strains of mice exhibited a VOMERONSASAL AND CHEMICAL
significant CPIR after oral stimulation, but not after post-
COMMUNICATION
oral stimulation with glucose. The fact that both strains
exhibited a CPIR of similar magnitude indicates that T1r3 Pre-absorptive insulin release to glutamate taste
is not necessary for eliciting the CPIR. In Experiment 2, we Matthew C Kochem1, Suzanne M Alarcon1, Paul AS Breslin1,2
measured blood glucose levels at 0, 15, 30, 60 and 120 min.
We found that both strains exhibited significantly better
1
Rutgers University New Brunswick, NJ, USA, 2Monell
glucose tolerance when the glucose was administered orally Chemical Senses Center Philadelphia, PA, USA
than post-orally; the benefit of oral administration was Glutamate, like glucose, requires insulin to be transported
especially pronounced in the Tas1r3 KO mice. These results out of blood into tissue. Glutamate is detected in the mouth
indicate that oral stimulation by sugars elicits a CPIR via by taste cells, in the intestine by L-cells and in the pancreas
a T1r3-independent taste mechanism, and that this CPIR by Beta cells. In addition, glutamate stimulates glucagon
substantially improves the ability of mice to tolerate glucose release from pancreatic alpha cells. It is unknown, how-
challenges. Acknowledgements: Summer Undergraduate ever, whether pre-absorptive insulin release (PIR) occurs
Research Fellowship Program, Columbia University in response to the taste of glutamate. The purpose of this
study was to determine whether glutamate causes PIR and a potential gender effect of this pathway on dietary fat
whether the magnitude of PIR was related to glutamate intake. Given these data, we have performed feeding studies
taste sensitivity, which is variable among individuals. To on female Trpm5−/− and Trpm5+/+ mice to look for similar
assess PIR, human subjects tasted and consumed a gluta- gender-specific effects on fat intake. Like male mice, female
mate test meal at a dose of 100mg/kg body weight. The glu- mice lacking Trpm5 show a decrease in dietary fat intake
tamate dose was comprised of a mixture of monosodium and gain less weight than wild types, though these effects
glutamate and monopotassium glutamate. Subjects tasted are much less robust and have a much slower onset than
the mixture by repeated swishing and spitting for 5 minutes. for the male mice. However, unlike the males, there is no
Subjects then ingested another dose of glutamate. Baseline significant difference in body fat between female Trpm5-/-
blood samples were collected every 5 minutes before inges- and Trpm5+/+ after being on the high fat diet for 56 days.
tion of glutamate. After ingestion, samples were collected We are currently exploring the specificity of these effects for
at 3 minute intervals for a total of 15 minutes and analyzed the different classes of fat (saturated versus unsaturated).
for glucose, insulin and c-peptide. To assess glutamate sen- Together, our data indicate that Trpm5 may play a role,
sitivity, subjects tasted and rated intensity matched NaCl, directly or indirectly, in the control of dietary fat intake
sucrose, and glutamate solutions. Relative glutamate sen- and that these effects appear to be significantly influenced
sitivity was defined as the ratio of perceived glutamate by gender. Acknowledgements: Supported by NIH grant
intensity to that of NaCl and sucrose. All seven subjects R01DK059611 (TAG)
showed pre-absorptive blood glucose changes in response
to the taste of glutamate. Six subjects showed increases in
blood glucose (presumably due to glucagon release) and one #P173 POSTER SESSION IV: CHEMICAL
subject showed a decrease. Half of the subjects showed pre- SIGNALING AND BEHAVIOR; ANIMAL
absorptive changes in insulin. The highest PIRs occurred BEHAVIOR/PSYCHOPHYSICS;
in the two subjects who showed the highest sensitivity to CHEMOSENSATION AND METABOLISM;
glutamate. The lowest PIR occurred in the subject with the VOMERONSASAL AND CHEMICAL
lowest sensitivity to glutamate. Acknowledgements: NIH COMMUNICATION
DC02995
Human Anticipatory Blood Pressure Responses to Oral NaCl
and KCl are Different
#P172 POSTER SESSION IV: CHEMICAL Melissa A Murphy1, Paul A.S. Breslin1,2
SIGNALING AND BEHAVIOR; ANIMAL 1
Department of Nutritional Sciences, Rutgers University
New Brunswick, NJ, USA, 2Monell Chemical Senses Center
CHEMOSENSATION AND METABOLISM; Philadelphia, PA, USA
COMMUNICATION The mechanism of association between dietary sodium and
blood pressure is not clearly defined, but high dietary salt
Mice lacking Trpm5 show reduced dietary fat intake intake is considered a risk for hypertension (HTN) and car-
Dulce M. Minaya, Jacqueline Lee, Dane R. Hansen, Timothy diovascular disease (CVD). Many studies have examined
A. Gilbertson the impact of different dietary cations on these risk factors
and implicate sodium (or Na/K ratio) as a key determinant.
Utah State University/Biology Department Logan, UT, USA
We previously presented data showing an anticipatory
We recently showed a critical role of Trpm5 in the trans- blood pressure (BP) response to the oral presentation of
duction pathway for long chain polyunsaturated fatty acids 1.0 Molar NaCl and suggested this may be linked to blood
(Liu et al., J Neurosci 31: 8634, 2011). In the present study, volume pre- absorptive reflexes. Based on this premise, we
we have begun to investigate dietary fat preference and the hypothesized that this anticipatory reflex would be more
propensity to develop dietary-induced obesity in mice lack- pronounced for sodium ions. To determine the specificity
ing Trpm5. In male mice placed on a high fat diet, those of the response to oral sodium, subjects rinsed orally with
mice lacking Trpm5 (Trpm5−/−) ate significantly less and either 0.5 M KCl (matched for taste intensity to NaCl) to
accordingly weighed less and had less body fat than wild test cation composition and 1.0 M Na-Gluconate to test
type (Trpm5+/+) mice; no differences between Trpm5−/− and salt taste intensity. Subjects rested in a seated position for
Trpm5+/+ mice were seen on a control (low fat) diet. Similar 2.5 hours while we recorded resting BP and additional
differences were recorded in control male mice and those readings following the rinse at 10 minute intervals with a
lacking IP3R3, receptors that are upstream of Trpm5 acti- manual sphygmomanometer, one trial per day, five trials
vation in the fatty acid transduction pathway. Most sur- for each solution tested. In subjects whose blood pressure
prisingly, however, was the fact that female mice with or dropped following a rinse with NaCl, a similar trend was
without IP3R3 did not show the same differences indicating observed after a rinse with Na-Gluconate. Yet BP remained
level, without a decreasing trend, following a rinse with #P175 POSTER SESSION IV: CHEMICAL
KCl. Interestingly, the BP response to Na-Gluconate was SIGNALING AND BEHAVIOR; ANIMAL
weaker than the NaCl response, suggesting an influence BEHAVIOR/PSYCHOPHYSICS;
of taste perception on the anticipatory BP reflex. The dif- CHEMOSENSATION AND METABOLISM;
ference between the BP response to NaCl and to KCl sug-
gests that there may be oral chemosensory specificity to
COMMUNICATION
the reflex. These data support the idea that anticipatory
BP reflexes are cation specific and may involve gustatory Effect of Oral Sensations on the Relief of Thirst
mechanisms. Acknowledgements: Funded in part by NIH Catherine Peyrot des Gachons1, Julie Avrillier2, Laure
DC 02995 to PASB. Algarra3, Emi Mura4, Paul A.S. Breslin1,5
1
#P174 POSTER SESSION IV: CHEMICAL 2
AgroSup Dijon Institut National Superieur Dijon, France,
AgroParisTech Paris, France, 4Suntory Business Expert
BEHAVIOR/PSYCHOPHYSICS; Ltd. Kawasaki, Japan, 5Rutgers University Department of
CHEMOSENSATION AND METABOLISM; Nutritional Sciences New Brunswick, NJ, USA
VOMERONSASAL AND CHEMICAL Thirst is the internal sensation of a need to drink, presum-
COMMUNICATION ably to rehydrate or recover bodily fluid losses. But thirst
Odor Identification Performance in Middle Aged Obese
is quenched long before all ingested fluids are absorbed.
Individuals with High Blood Pressure
Therefore, sensory feedback must play a role in thirst
quenching. Different beverages seem to quench thirst with
Stephanie M. Oleson1, Erin Green3, Aaron Jacobson3, Claire different efficiencies, but how their oral sensory charac-
Murphy1,2,3 teristics determine the thirst quenching efficacy is poorly
1
San Diego State University San Diego, CA, USA, 2University understood. The purpose of this study was to determine
of California, San Diego San Diego, CA, USA, 3SDSU-UCSD which oral sensation(s) commonly manipulated in bever-
Joint Doctoral Program in Clinical Psychology San Diego, CA, ages, such as temperature or carbonation, influence lev-
USA els of thirst. To answer this question, subjects who were
deprived of liquid overnight were first asked to drink a
Obesity is a major health epidemic that affects people fixed volume of an experimental beverage presenting one
globally and is associated with the development of seri- or two specific traits. Then we objectively evaluated their
ous comorbidities such as diabetes, hypertension, and residual thirst by measuring how much additional plain,
heart disease. The presence of obesity has also been linked uncarbonated, room temperature water they wanted to
to the risk for later development of Alzheimer’s Disease drink afterwards. The results show that the perception of
(AD) or mild cognitive impairment (MCI) and has also coldness is an important parameter for thirst quenching.
been associated with poorer performance on cognitive A beverage at low temperature (5°C) quenches thirst more
measures of global, executive, and memory functioning. than a beverage at room temperature (20°C). Moreover, a
Furthermore, olfactory functioning is linked to energy bal- cold, carbonated beverage relieves thirst even more than
ance and metabolism and has been shown to be altered in does a cold uncarbonated beverage. These results support,
obese individuals, as well as those diagnosed with AD or in part, the observations of the sensory controls of thirst
MCI. It has been proposed that the impact of obesity on quenching in the animal literature. Acknowledgements:
cognition is indirect and influenced by comorbidities such Suntory Business Expert Ltd.
as hypertension and diabetes. Therefore, the current study
sought to determine if odor identification performance
differed between obese individuals with and without high
blood pressure. Thirty- one obese individuals (BMI > #P176 PPOSTER SESSION IV: CHEMICAL
30 kg/m2) between the ages of 46–54 years old were given SIGNALING AND BEHAVIOR; ANIMAL
the San Diego Odor Identification test. Obese individuals BEHAVIOR/PSYCHOPHYSICS;
with high blood pressure performed significantly worse on CHEMOSENSATION AND METABOLISM;
the odor identification task (F= 8.384; p = .008), but nei- VOMERONSASAL AND CHEMICAL
ther group significantly differed in BMI or odor threshold. COMMUNICATION
The results suggest that obese individuals with high blood
pressure are more likely to show odor memory deficits and Intraduodenal infusions of sucrose influence conditioned
that these changes occur as early as during middle-age. and unconditioned affective taste-guided responses to oral
Acknowledgements: Supported by NIH grant #AG004085- sucrose
25 to CM. Lindsey A. Schier, Alan C. Spector
Department of Psychology and Program in Neuroscience, human studies have generated conflicting results. This study
Florida State University Tallahassee, FL, USA tested whether the influence of acute homeostatic state
Sensory signals ascending from the oral cavity and viscera (fasted vs. fed) on olfactory perception could be moderated
are integrated in the CNS to adjust meal size and taste by body mass index, which is associated with chronic meta-
preferences. Electrophysiological data suggest such integra- bolic alterations. Twenty-one subjects (14 healthy weight/
tions affect early processing in brainstem taste nuclei. Taste HW, BMI M=22.3 SD=1.9; 7 overweight/OW, BMI M=28.2
receptors (e.g. T1R) are also found in GI cells, but whether SD=2.8) rated the intensity of odors (chocolate, strawberry,
taste-like signals arising from postoral sites are integrated honeysuckle and lilac) and tastes (chocolate and strawberry
with oral taste-guided behaviors is unknown. This study flavored milk) using the general labeled magnitude scale in
examined effects of intraduodenal (ID) infusions of a sweet fed and fasted states (separate counterbalanced days), con-
ligand on conditioned and unconditioned affective responses comitant to an fMRI study. Blood samples were also col-
to matching oral chemosensory stimuli. First, rats were given lected on a subset of subjects (n=15) to assess the influence of
two separate 15 min sessions to ingest 0.3M sucrose directly internal state on feeding-related molecules. A repeated meas-
followed by either ip LiCl (3 mEq/kg) to condition a sucrose ures ANOVA of intensity ratings revealed a 3-way interac-
taste aversion (CTA, n=9) or saline (unconditioned, n=8). tion between state (fed vs. fasted), stimulus (odors vs. taste)
Then, licking responses to 5 sucrose concentrations (0.01, and group (HW vs. OW) (f(1,19)=7.52, p=.013), where odor
0.03, 0.1, 0.3, 1M), 0.12M NaCl, and water (10s trials in ran- intensity ratings decreased significantly in fed vs. fasted for
domized blocks) were examined in two 30 min brief-access OW but not HW subjects. Critically, the difference in odor
tests. Four min before each test, rats were infused ID with intensity ratings between fasted and fed correlated negatively
0.3M sucrose or 0.15M NaCl (3ml). For unconditioned rats, with difference in ghrelin change from baseline between
ID sucrose enhanced preferential licking to 0.03–1 M oral fasted and fed conditions (r(13)=−.615, p=.016); the smaller
sucrose, with no effect on licking for NaCl. This preference the change in intensity perception, the larger the change in
shift emerged rapidly by trial block 2. CTA rats reduced ghrelin levels. These findings demonstrate that chronic and
licking to 0.03−1M sucrose, not NaCl, but because CTA acute metabolic states interact to influence olfactory percep-
rats initiated so few trials, and significantly fewer after ID tion possibly through ghrelin signaling. Acknowledgements:
sucrose versus ID NaCl, detection of emergent differences Supported by R01 DK085579.
due to ID preload type was likely limited. Thus, a taste
reactivity test was conducted in a separate group of CTA
rats. ID 0.3M sucrose preload (n=6) significantly increased
gaping to intraoral 0.3M sucrose infusions (0.5ml/30s every
3min for 15min) relative to ID 0.15M NaCl preload (n=7). BEHAVIOR/PSYCHOPHYSICS;
Together, the results suggest postoral stimuli impact oral CHEMOSENSATION AND
taste processing in chemospecific ways. Acknowledgements: METABOLISM; VOMERONSASAL AND
NIH-T32DC000044 CHEMICALCOMMUNICATION
Oral Sweet Taste Stimulation Induces Cephalic Phase
#P177 POSTER SESSION IV: CHEMICAL Carbohydrate Oxidation in Humans
SIGNALING AND BEHAVIOR; ANIMAL Sze Yen Tan, Richard D. Mattes
BEHAVIOR/PSYCHOPHYSICS; Purdue University/Department of Nutrition Science West
CHEMOSENSATION AND METABOLISM; Lafayette, IN, USA
COMMUNICATION Objective: Sensory stimuli induce many anticipatory reflexes.
Specifically, oral sweet taste stimulation increases glucose
An interaction of chronic and acute metabolic states on absorption in rats and induces cephalic phase insulin release
olfactory perception associated with ghrelin signaling in animals and humans. This study aimed to investigate
Xue Sun, Marga G Veldhuizen, Dana M Small whether oral sweet taste stimulation also induces an acute
increase of carbohydrate oxidation in humans. Methods:
The John B. Pierce Laboratory and Yale University New This was a randomized, crossover design study with two test
Haven, CT, USA visits delivering: 1) control (DI water) and 2) 10% sucrose
Olfaction is key for the evaluation of food cues in the (w/w) solutions orally. An indirect hood calorimeter was
environment. It is therefore not surprising that evidence is used to measure energy expenditure (EE) and respiratory
emerging for metabolic influences in olfaction. For example, quotient (RQ) through gaseous exchanges. Each measure-
receptors for feeding-related molecules such as ghrelin are ment was performed at one to two hours after habitual
expressed within the olfactory system and olfactory sensitiv- breakfast or lunch, where participants were instructed to
ity is increased in fasted vs. fed state in rodents. However, consume an identical meal at identical times before their
visits. Calorimeter measurement was preceded by a 30-min- glucose transporter. We demonstrated by immunocyto-
ute habituation period, followed by oral stimulation and sub- chemistry the ciliary location of this transporter in isolated
sequent measurement for 30 minutes. Changes in EE and RQ rat and toad olfactory neurons. Additionally, field record-
were tested using a general linear model for repeated measures ings (electroolfactogram) indicated that inhibition of gly-
ANOVA in SPSS. Results: Nine participants have completed colysis and oxidative phosphorylation impairs the odor
the study (8 females and 1 male, mean age=25.2 ± 4.6 years, response. Altogether, these results are consistent with a dual
mean weight=59.2 ± 4.3kg, mean BMI=21.6 ± 2.5kgm−2). supply of ATP in olfactory cilia, oxidative phosphorylation
Energy expenditure following test solution stimulation and glycolysis. Acknowledgements: FONDECYT 1100682
did not change and was not significantly different between (JB), DI/VRIEA/PUCV (JGR)
solutions (time and interaction effects, P>0.05). However,
RQ was significantly increased after sweet taste stimulation
(time effects, p<0.001; interaction effects, p=0.018), which #P180 POSTER SESSION IV: CHEMICAL
occurred mainly during the first 10 minutes after exposure. SIGNALING AND BEHAVIOR; ANIMAL
Conclusions: Oral sweet taste stimulation increased carbo- BEHAVIOR/PSYCHOPHYSICS;
hydrate oxidation in humans. The specificity of the response CHEMOSENSATION AND METABOLISM;
remains to be determined. VOMERONSASAL AND CHEMICAL
COMMUNICATION
#P179 POSTER SESSION IV: CHEMICAL Dietary calcium intake and ethnicity may contribute to
SIGNALING AND BEHAVIOR; ANIMAL individual differences in taste perception.
BEHAVIOR/PSYCHOPHYSICS; Anna Voznesenskaya, Laura K. Alarcón, Michael G. Tordoff
CHEMOSENSATION AND METABOLISM; Monell Chemical Senses Center Philadelphia, PA, USA
COMMUNICATION Calcium status affects preferences for calcium and sweet solu-
tions in rats and mice: calcium deficient rodents drink more
Energy supply in chemosensory cilia of olfactory receptor calcium solutions and avoid sweet compounds. Moreover,
neurons: Possible role of glycolysis preference for calcium is inversely correlated with preference
Pablo S Villar1,2, Juan G Reyes1, Juan Bacigalupo2 for sweet compounds in calcium replete mice. However, lit-
tle is known about the relationships between calcium status
1
Instituto de Química, Facultad de Ciencias, Pontificia
and taste perception in humans. Here we measured detection
Universidad Católica de Valparaíso Valparaíso, Chile,
thresholds for CaCl2 and sucrose, and assessed intensity and
2
Departamento de Biología, Facultad de Ciencias,
taste quality ratings of CaCl2, sucrose, NaCl, QHCl and cit-
Universidad de Chile Santiago, Chile
ric acid in an ethnically diverse group of people in relation to
The chemosensory cilia of olfactory sensory neurons are dietary calcium intake. African-Americans had significantly
long and thin structures (60 x 0.2 µm) devoid of inner higher detection thresholds than Caucasians for both CaCl2
membranes, specialized in odorant transduction. A cAMP and sucrose. They rated 25 mM CaCl2 as predominantly
pathway couples the activation of odor receptors with the sour significantly more frequently than Caucasians. There
opening of cyclic nucleotide-gated channels. During the was no relationship between dietary calcium intake and
odor response, the cilia undergo high levels of ATP hydroly- CaCl2 detection threshold. In African- Americans but not
sis, as this nucleotide is used by adenylyl cyclase, ATPases Caucasians sucrose detection threshold was inversely corre-
and kinases. Our estimates of resting ATP level, ATP dif- lated with dietary calcium levels (Spearman’s rho = −0.93,
fusion and consumption suggest that the mitochondria, p<0.001). Dietary calcium levels also affected psychophysi-
located near the base of the cilia, are insufficient to sustain cal ratings of NaCl and citric acid. Ethnic differences in
chemotransduction in the entire cilium under intensive stim- taste perception are often explained by socioeconomic fac-
ulation. Nuñez-Parra et al (Chem Senses 36:771-7802012) tors but there are large disparities in calcium metabolism
found glucose transporters in the sustentacular cells of the and genetic variations in sweet taste receptor genes between
olfactory epithelium. We hypothesize these cells release glu- African-Americans and Caucasians that may underlie the
cose to the mucus, the cilia incorporated it from there and racial differences observed in our study. Our results provide
utilize it by glycolysis, supplementing the required ATP. To new information about the relationship between calcium
test this idea, we detected glycolytic enzymes by immuno- status and taste perception in humans; they underscore
blot of a ciliary membrane preparation. Additionally, we the importance of further investigation into the metabolic
measured cilia and knob accumulation of a fluorescent mechanisms responsible for ethnic differences in taste per-
deoxyglucose analog when applied to mucosal side of the ception. Acknowledgements: Pennsylvania Health Research
olfactory epithelium, suggesting the apical presence of a Formula Funds
#P181 POSTER SESSION IV: CHEMICAL Ali Magableh, Robert Lundy

SIGNALING AND BEHAVIOR; ANIMAL University of Louisville School of Medicine/Anatomical
BEHAVIOR/PSYCHOPHYSICS; CHEMOSENSATION Sciences and Neurobiology Louisville, KY, USA
AND METABOLISM; VOMERONSASAL AND Learning plays a crucial role in the establishment and
CHEMICAL COMMUNICATION strengthening of food preference and we hypothesize that
Differences in food cue reactivity between normal weight specific limbic system neuropeptide pathways play an
and overweight individuals? important role. Identifying neural mechanisms that medi-
Harriët F.A. Zoon1, Wei He1,2, Rene A. de Wijk2, Cees de ate affective aspects of taste perception will further our
Graaf1, Sanne Boesveldt1 understanding of how the brain controls eating and over-
eating. We have identified two neuropeptides, corticotrophin
1
Division of Human Nutrition, Wageningen University
releasing factor (CRF) and somatostatin (Sst), which are
Wageningen, Netherlands, 2Food and Biobased Research,
expressed in limbic system neurons that project to a hind-
Wageningen UR Wageningen, Netherlands
brain neural substrate critical for establishment of gustatory
Overweight occurs when energy intake exceeds energy expend- hedonic value; the pontine parabrachial nucleus. Our goal is
iture over the long term. Overweight people have been sug- to develop a viral construct capable of directing conditional
gested to be more sensitive to rewarding effects of food (e.g. expression of nitroreductase gene (NTR) to Sst and CRF
Davis et al., 2004; Franken & Muris, 2005). In the anticipatory cell populations in the limbic system of mice using a cre/lox
phase of eating, odors can be considered as external cues that system. Thus, specific peptide producing neurons can be rap-
signal the energy content and hence the reward value of a food. idly ablated in isolation following treatment with the prod-
In overweight individuals internal hunger signals are thought rug CB1954 allowing assessment of their role in central taste
to be overruled by external food cues (Herman & Polivy, 2008). processing and taste-guided behaviors. In vitro cell culture
This study aims to determine if food cue reactivity is higher in of HEK293 cells combined with FLOW cytometric analysis
overweight compared to normal weight individuals. Frequency indicate that we can conditionally express NTR and cause
of choice for energy-dense food items and amount of food cell death following CB1954 treatment. Acknowledgements:
intake reflect food cue reactivity. 25 overweight (BMI mean: This research work was supported by a grant from the
31.33 kg/ m2, SD: 3.36) and 25 normal weight (BMI mean: Kentucky Science and Engineering Foundation as per Grant
21.84 kg/ m2, SD: 1.78) females, matched on age and restraint Agreement #KSEF-148-502-11-277 with the Kentucky
score, participated. In 6 separate sessions they were exposed to Science and Technology Corporation.
odors of three different categories (signaling non-food, high-
energy food, low-energy food) in two motivational states (hun-
gry and satiated). High-energy preference was measured with
a computerized forced choice task and food intake (kCal) was #P183 POSTER SESSION IV: CHEMICAL
determined with the use of a Bogus Taste Test. We hypothesize SIGNALING AND BEHAVIOR; ANIMAL
that increased food cue reactivity in overweight women is dem- BEHAVIOR/PSYCHOPHYSICS;
onstrated by a stronger tendency to choose high-energy food CHEMOSENSATION AND METABOLISM;
products after being exposed to high-energy food odors, and VOMERONSASAL AND CHEMICAL
may subsequently lead to more food intake compared to lean COMMUNICATION
individuals. However, preliminary results (N=28) indicate that
Combinatorial and genotype specific co-expression of the
there is no main effect of odor on high-energy food preference
major urinary proteins (MUPs) during mouse postnatal
(p=0.755) and also no interaction effect between odor and BMI
development: from fundamental aspects of olfaction to
group (p=0.935). Our first results on food intake (N=48) indi-
innovative prospects in biomedicine
cate no main effect of odor (p= 0.792) and no interaction effect
of odor and BMI group (p=0.323). Acknowledgements: This Sergey N. Novikov1, Elena M. Fedorova1,2, Irina I. Ermakova3,
study was funded by NWO (The Netherlands Organization for Anatoly A. Philimonenko4, Gennady A. Churakov5, Sergey
Scientific Research), Veni grant nr. 451-11-021, awarded to SB. V. Mylnikov6
1
I.P. Pavlov Institute of Physiology, Russian Academy of
#P182 POSTER SESSION IV: CHEMICAL Sciences Saint Petersburg, Russia, 2Institute of Experimental
Medicine, Russian Academy of Medical Sciences Saint
Petersburg, Russia, 3Institute of Cytology, Russian Academy
BEHAVIOR/PSYCHOPHYSICS; CHEMOSENSATION
of Sciences Saint Petersburg, Russia, 4Institute of Molecular
AND METABOLISM; VOMERONSASAL AND Genetics, v.v.i., Academy of Sciences of the Czech Republic
CHEMICAL COMMUNICATION Prague, Czech Republic, 5Institute of Experimental
Development of Viral Based Gene Delivery for Conditional Pathology/Molecular Neurobiology (ZMBE), University
Ablation of Specific Brain Peptidergic Neurons of Muenster Muenster, Germany, 6Saint Petersburg State
University, Department of Genetics and Biotechnology Saint Epidemiology & Community Health Minneapolis, MN,
Petersburg, Russia USA, 5University of Minnesota, Department of Laboratory
Medicine & Pathology Minneapolis, MN, USA
Major urinary proteins (MUPs) of the house mouse form
a large group of highly polymorphic acidic isoforms with Taste sensation may influence food choice and consumption
molecular masses of 18–20 kDa. MUPs are encoded by the which in turn may play a role in the maintenance of health.
Mup gene cluster, which consists of about 35 genes and pseu- The objective of this study was to determine the relationship
dogenes and is mapped to chromosome 4. Nowadays MUPs between taste and changes in adiposity and related health
are considered as a key component of the mouse olfactory measures during a 5 year follow-up period in the Beaver
signature which can provide all essential information about Dam Offspring Study. Whole mouth suprathreshold taste
the individuality of donors. There is also rapidly growing evi- intensity was measured for salt, sweet, sour, and bitter at
dence that several MUP isoforms are involved in the regula- baseline (2005–2008) using filter paper disks and a general
tion of glucose metabolism (Zhou, Rui, 2010) and can be used labeled magnitude scale. Health outcomes measured at base-
as sensitive biomarkers in early diagnosis of experimental line and follow-up (2010–2013) included body mass index
nephritis (Wenderfer et al., 2009) and hepatocarcinogenesis (BMI), waist circumference, systolic and diastolic blood
(Ritorto, Borlak, 2011). These studies open practically unex- pressure, total cholesterol, and hemoglobin A1c (HbA1c).
plored biomedicine avenue for using MUPs as new protein Cluster analysis was used to group participants according
markers which are very suitable for diagnostic purposes. We to observed patterns of intensities of the 4 tastes. In prelimi-
examined ontogenetic profiles of MUPs expression in male nary analyses (n = 1681, mean age at baseline = 48.9 years,
and female mice of CBA/LacY and C57BL/6JY strains using range = 22–84 years), there were associations between pat-
electrophoresis in polyacrylamide gel (PAGE). Quantitative terns of taste intensities and 5-year changes in BMI, waist
evaluation of eight MUPs isoforms (A-H) revealed that each circumference, and HbA1c level. With adjustment for age
genotype is characterized by specific combinations and differ- and sex, the cluster with high intensities for all 4 tastes
ent proportions (ratios) of the same MUP fractions. These sex demonstrated a significantly greater mean increase in BMI
and genotype specific ratios emerged in both sexes very soon (+ 0.96 kg/m2) and HbA1C (+ 0.37%) than the cluster with
after weaning, remain quite constant in adults and resemble average intensities for the 4 tastes (BMI: + 0.32 kg/m2;
«barcode». Our data suggest that the pattern of Mup genes HbA1c: + 0.21%). Similar results were observed for waist
expression during mouse ontogenesis is regulated through circumference (high intensities cluster: + 3.01 cm; average
a very stable genetic program. We suppose that at the early intensities cluster: + 1.87 cm). In these preliminary analy-
stage of illness this ontogenetic program is destroyed and ses, oral sensation, characterized using patterns of perceived
the expression pattern of several Mup genes will be changed. intensities of suprathreshold tastes, was found to be associ-
These processes are reflected in the appearance of new pro- ated with 5-year changes in some adiposity- related health
tein profiles with altered MUPs’ ratios and may correspond outcomes. Acknowledgements: The project described was
to epigenetically changed expression of the Mup gene cluster. supported by R01AG021917 from the National Institute
Acknowledgements: Supported by Russian Foundation for on Aging, National Eye Institute, and National Institute on
Basic Research (projects 02-04-49273, 07-04-01762). Deafness and Other Communication Disorders. The content
is solely the responsibility of the authors and does not neces-
sarily reflect the official views of the National Institute on
#P184 POSTER SESSION IV: CHEMICAL Aging or the National Institutes of Health.
BEHAVIOR/PSYCHOPHYSICS; #P185 POSTER SESSION IV: CHEMICAL
CHEMOSENSATION AND METABOLISM; SIGNALING AND BEHAVIOR; ANIMAL
VOMERONSASAL AND CHEMICAL BEHAVIOR/PSYCHOPHYSICS;
COMMUNICATION CHEMOSENSATION AND METABOLISM;
The Association of Taste with Adiposity in the Beaver Dam VOMERONSASAL AND CHEMICAL
Offspring Study COMMUNICATION
Mary E. Fischer1, Karen J. Cruickshanks1,2, Carla R. Schubert1, Social Olfactory Cues and Stress
Guan-Hua Huang3, Barbara E.K. Klein1, Ronald Klein1, James
Pamela Dalton, Cristina Jaen, Tamika Wilson,
S. Pankow4, Nathan Pankratz5, Alex Pinto1
Christopher Maute
1
University of Wisconsin, Department of Ophthalmology &
Visual Sciences Madison, WI, USA, 2University of Wisconsin,
Department of Population Health Sciences Madison, WI, Olfactory cues have the potential to precipitate emotional
USA, 3National Chiao Tung University, Institute of Statistics responses and thereby alter mood, judgments and behavior.
Hsinchu, Taiwan, 4University of Minnesota, Division of In prior work, we demonstrated that exposure to a novel
odor while undergoing a laboratory stressor caused indi- to the subject’s own body odor (Self). Using event-related
viduals to re-experience stress (increased heart rate & self- potential (ERPs), we have more recently demonstrated that
reported stress) when re-exposed to that odor three days later. the presence of a Strangers’ body odor causes a non-threat-
We wished to evaluate the potential for odor to alter stress ening face to be processed as a threatening stimulus. In the
levels following a standard laboratory stressor. One class of present ERP study, we sought to determine whether the che-
olfactory stimuli which has shown promise in eliciting robust mosignal mediating the aforementioned ERP effect could
effects on mood and emotion are social odors. Lundström be masked by a common odor (Mask). Angry and neutral
et al. (2008) demonstrated that smelling a stranger’s body schematic faces were presented to subjects in the presence of
odor activated a marked response in the amygdala of sub- Strangers’ body odor + Mask, ‘Self ‘ body odor + Mask, or
jects, despite a low conscious recognition of the odor or its Mask only control, which were delivered intra-nasally by a
source. In this study, we evaluated the changes in autonomic computer-controlled olfactometer. Preliminary analyses sug-
stress levels following the Trier Social Stress Test among gest that even in the presence of an odor mask, exposure
individuals in 3 groups who were exposed to either the body to the body odor of a stranger, relative to the odorless con-
odor of their sibling, the body odor of a stranger or a non- trol and ‘Self ‘ body odor, results in significant differences
social (fragrance) odor. Axillary odors were collected from in the late (cognitive) components of visual processing. This
non-twin, whole, biological siblings who were then recalled suggests that body odor can modulate the cognitive evalu-
to participate in the main study in which one of the 3 odors ation of visual stimuli even in the presence of a perceptual
was administered following the stress task. Results showed a odor mask. The effects of masked body odor and common
significant decrease in post-recovery heart rate only among odor exposure on visual processing will be presented and
the group smelling the sibling odor, whereas skin conduct- discussed within the framework of the adaptive advantages
ance was significantly reduced for both the sibling odor and conveyed by heightened sensitivity to threat-related stim-
the fragrance. Following a stressor, exposure to the stranger uli. Acknowledgements: This work was supported by the
body odor maintained arousal levels longer suggesting that National Institute on Deafness and other Communication
both familiar and stranger body odors may be potent cues Disorders – NIDCD (R03DC009869) awarded to JNL
for emotional responses. Acknowledgements: Supported by
the U.S. Army Research Office grant # W911NF-11-1-0087,
entitled “Learning & Olfaction: Understanding and #P187 POSTER SESSION IV: CHEMICAL
Enhancing a Critical Communication Channel”. SIGNALING AND BEHAVIOR; ANIMAL
Differential responses to two kairomonal cues in
mosquitoes
COMMUNICATION Shahid Majeed, Sharon R. Hill, Teun Dekker, Göran
Birgersson, Rickard Ignell
Can a chemosensory threat be masked?
Swedish University of Agricultural Sciences Alnarp, Sweden
Amy R Gordon1,2, Kathrin Ohla2,3, Mats J Olsson1, Johan N
Lundstrom1,2,4 Culex quinquefasciatus, Aedes aegypti, and Anopheles gam-
biae are vectors of diseases that are among the main causes
1
Karolinska Institutet Stockholm, Sweden, 2Monell Chemical
of human mortality and morbidity worldwide. Host-seeking
Senses Center Philadelphia, PA, USA, 3German Institute of
in these species is primarily regulated by olfactory cues. The
Human Nutrition Potsdam-Rehbrücke, Germany, 4University
most important cue for the activation of host-seeking is car-
of Pennsylvania Philadelphia, PA, USA
bon dioxide (CO2), the principal by-product of respiration.
The well-established angry advantage effect has been All three mosquito species were able to detect and follow
extended in recent crossmodal visual-olfactory studies using pulsed stimuli of CO2 at the level of olfactory receptor neu-
schematic human faces and human body odors. In a threat- rons (ORNs) housed within capitate pegs on the maxillary
detection task, humans detect an angry (threatening) sche- palps. The temporal coding capacity of C. quinquefasciatus
matic face in an array of neutral distracter faces more quickly CO2-sensitive ORNs, however, was significantly lower than
than a friendly (non-threatening) face, hence the label ‘Angry that of the other two species. This differential physiological
advantage’. We have previously shown that the body odor of response was reflected in the behavioral response to CO2, and
unknown individuals (Strangers) – an established threatening correlates with the CO2 emissions from the preferred hosts for
olfactory stimulus – speeds a subject’s detection of threaten- each of these species. Furthermore, aeration extracts taken
ing faces, but not non-threatening faces, relative to exposure from preferred hosts were analyzed by gas chromatography
coupled single sensillum recording (GC-SSR) of the capitate Katharine A. Prokop-Prigge1, George Preti1,2
pegs. We identified (R)-1-octen-3-ol, a component in human 1
headspace volatiles, as a physiologically active in each spe- 2
University of Pennsylvania, School of Medicine, Department
cies, although with different sensitivities. It is interesting to of Dermatology Philadelphia, PA, USA
note that (R)-1-octen-3-ol was absent from bird aeration
extracts. Landing bioassays using the host aeration extracts The human adenosine triphosphate (ATP)-binding cassette
revealed behavioral responses of the three species consistent (ABC) transporter ABCC11 gene encodes an ATP-driven
with their host selection preferences. The addition of bio- efflux pump protein that plays an important function in
logically relevant concentrations of (R)-1- octen-3-ol to bird ceruminous glands of the auditory canal. This protein is
aeration extracts either inhibited or increased the behavioral also expressed in apocrine sweat glands and appears to reg-
response of the mosquitoes, consistent with its role as a non- ulate the secretion of axillary odor precursors. It has been
host and host volatile, respectively. Here, we show that the shown that a simple change in the ABCC11 gene results in
host-seeking behavior of mosquitoes may be differentially different types of axillary odorant and ear wax production.
regulated by olfactory signals emitted by potential hosts in Individuals who are homozygotic for a single nucleotide
their environment. polymorphism (538G>A), which changes amino acid 180
in the resultant protein’s polypeptide chain from glycine (G)
to arginine (R), were found to have significantly less of the
#P188 POSTER SESSION IV: CHEMICAL characteristic axillary odorants than either those who are
SIGNALING AND BEHAVIOR; ANIMAL heterozygotic for this change or those who had the wild type
BEHAVIOR/PSYCHOPHYSICS; gene. Asian populations differ markedly from non-Asians in
their ear wax type and underarm odor production. G180R
SNP also is associated with a dry, white earwax phenotype
that is predominant in Asians. The G180R SNP is rare in
COMMUNICATION Africans and Caucasians who typically exhibit wet, yellow
Effects of 5a-Androst-16-EN-3a-OL Scent Administration on earwax. For the first time, analytical analysis of earwax odor-
Augmenting Male Gambling Behavior ants has been performed and the principle odorants in both
earwax phenotypes will be discussed. The odor of each ear
Sierra Moore, Bryan Raudenbush, Patrick Dwyer
wax type was informally accessed and the principal odorants
Wheeling Jesuit University Wheeling, WV, USA were found to be volatile organic C2 -to-C8 acids. A compari-
The effects of pheromones on non-humans have been exten- son between volatile earwax and axillary odors will also be
sively studied. However, less is known on how such scents can presented. Acknowledgements: NIH postdoctoral training
elicit responses in humans. Previous research indicates that the grant (2T32DC000014-32A1) ARO (W911NF-11-1-0087)
scent of a female can affect various emotions and behaviors of
males. The present study assessed the effects of Androstenol
on gambling behavior in males. Participants completed an #P190 POSTER SESSION IV: CHEMICAL
ad libitum blackjack gambling task while exposed to either SIGNALING AND BEHAVIOR; ANIMAL
no scent (control) or androstenol (experimental condition). BEHAVIOR/PSYCHOPHYSICS;
Results indicate males gambled for a significantly longer period CHEMOSENSATION AND METABOLISM;
of time when exposed to androstenol, t(36) = 2.09, p<.05. VOMERONSASAL AND CHEMICAL
Scores on a Sensation Seeking Scale also indicate an interac- COMMUNICATION
tion between scent administration and personality character-
istics. Thus, the administration of certain female pheromones Loss and Recovery of Odorant-Mediated Behavior
may lead to an increased desire in males to engage in sensation Correlates with Plasticity of Axonal Projections
seeking or risk-taking behaviors Acknowledgements: West in the Zebrafish Olfactory Bulb in a Reversible
Virginia Space Grant Undergraduate Research Fellowship Deafferentation Model
Evan J White, Taylor R Paskin, Christine A Byrd-Jacobs
Western Michigan University/Biological Sciences Kalamazoo,
MI, USA
BEHAVIOR/PSYCHOPHYSICS; We have found that the repeated exposure of adult zebrafish
CHEMOSENSATION AND METABOLISM; olfactory epithelium to the detergent Triton X-100 results
in fish losing the ability to respond to odorants associated
with social behavior but retaining the ability to respond
COMMUNICATION
to odorants linked to feeding behavior. Using a reversible
Earwax and Underarms; the Odd Odor Couple deafferentation technique, we find that fish recover the
ability to detect social cues. The aim of the present study of behaviorally relevant chemosignals: major urinary pro-
was to determine a biological basis for this phenomenon teins (MUPs). Using extracellular ‘loose-seal’ patch-clamp
by examining axonal projections after a single treatment recordings from optically identified basal VSNs in acute
with TX-100. Axons of three olfactory sensory neuron coronal VNO slices, we record stimulus-dependent action
subtypes (ciliated, microvillar, and crypt) were identified potential discharge in response to recombinant MUPs,
using immunocytochemistry on paraffin sections. In control specific for either the C57BL/6J or the BALB/cByJ inbred
bulbs, anti-KLH labeled all glomeruli, while anti-calretinin strain of laboratory mice. Furthermore, we comparatively
labeled fewer axons throughout the bulb. Anti-Gas/olf labe- analyzed the role(s) of these stimuli in two different male-
ling was concentrated in the medial and dorsal bulb, and specific behaviors: male-male aggression and territorial
anti-S-100 labeling was more obvious in the lateral bulb. countermarking. Surprisingly, electrophysiological activ-
Within the first 4 days after TX-100 treatment, anti-KLH ity profiling revealed parallel detection of MUPs by both
and anti-calretinin labeling in the deafferented bulb showed ‘specialist’ neurons selectively tuned to a particular stimu-
an overall reduction, with prominent loss in the medial lus and broad range responders (‘generalists’) sensitive to
bulb and preservation of some axons in the lateral bulb. By all or subset combinations of the MUPs tested. These data
7 days, innervation returned to near control levels. Staining suggest the coexistence of determined and variable sensory
in the deafferented bulb with anti-Gas/olf and anti-S-100 coding strategies in the mouse vomeronasal system. In
was absent 1 day following treatment but returned within addition, behavioral assays indicate that MUPs regulate at
7 days. Examination of the axon patterns showed a selective least two different male behaviors. While dedicated ligands
preservation of certain olfactory sensory axons, while oth- promote aggression, a combinatorial MUP code controls
ers are temporarily destroyed. The presumptive microvillar countermarking. Together, our results show that a vomer-
axons that survive treatment in the lateral bulb may account onasal stimulus can encode divergent information through
for the persistent ability of zebrafish to detect food odor- both dedicated and combinatorial neural mechanisms.
ants while the temporary destruction of ciliated axons in the
medial bulb is consistent with the loss and recovery of the
ability to detect social cues. Acknowledgements: Supported #P192 POSTER SESSION IV: CHEMICAL
by NIH-NIDCD #011137 (CBJ) SIGNALING AND BEHAVIOR; ANIMAL
Insights into the Function of Darcin from the Three
Dimensional Structure
COMMUNICATION Robert J Beynon, Marie M Phelan, Lu-Yun Lian, Lynn
McLean, Jane L Hurst
Coexistence of determined and variable sensory coding
strategies in the mouse vomeronasal system University of Liverpool / Institute of Integrative Biology
Liverpool, United Kingdom
Tobias Ackels1, Annika Cichy1, Angeldeep Kaur2, Maria
Kateri3, Tobias Marton2, Darren Logan2,4, Lisa Stowers2, Marc Mouse urine contains millimolar concentrations of major
Spehr1 urinary proteins (MUP) - eight stranded beta barrel lipoca-
lins. MUPs have a broad range of functions in chemical
1
Department of Chemosensation, RWTH Aachen University
communication between individuals. One MUP, darcin
Aachen, Germany, 2Department of Cell Biology, The Scripps
(MGI classification Mup20) is highly expressed in males
Research Institute La Jolla, CA, USA, 3Institute of Statistics,
only and is responsible for inherent female attraction to
RWTH Aachen University Aachen, Germany, 4Wellcome Trust
males, the subsequent memory of the volatile odour pro-
Sanger Institute, Hinxton Cambridge, United Kingdom
file of that male and the rapid induction of memory of
The mouse vomeronasal organ (VNO) is an important the precise physical placement of the scent mark contain-
chemosensory subsystem that has been implicated in a ing darcin. Darcin is responsible for the slow release of
variety of social and sexual behaviors. In contrast to com- the volatile pheromone, 2-sec-butyl 4,5 dihydrothiazole.
binatorial odor coding by neurons in the main olfactory To better understand the unique properties of darcin, we
epithelium, vomeronasal sensory neurons (VSNs) are have solved the structure of this protein by NMR. Relative
thought to function as narrowly tuned, dedicated sensors to other MUPs, darcin has a large solvent exposed area
of intrinsically instructive semiochemicals. Here, we inves- and the greatest exposure of hydrophobic residues in the
tigate the tuning profile(s) of a group of VSNs that are col- beta barrel. Binding to three ligands – NPN, menadione
lectively characterized by their sensitivity to a specific class and a thiazole derivative – were characterized in this study.
Menadione and thiazole bound to both darcin and MUP11 proton-induced responses indicate a critical role of hyper-
in the hydrophobic cavity of the beta barrel, with NMR polarization-activated cyclic-nucleotide- gated (HCN)
data indicating a similar binding site for menadione and ion channels in proton-mediated signaling of vomerona-
thiazole in both darcin and MUP11. The largest ligand sal sensory neurons. Together, our results implicate HCN
(NPN) bound only to MUP11, not darcin, suggesting dar- channel-dependent vomeronasal acid-sensing in gain con-
cin adopts a more compact binding cavity. Darcin is sig- trol of social chemosignaling.
nificantly more stable than MUP11, with 92% of darcin
structure retained in the native state at 7M urea compared
to only 45% in MUP11. The high stability of darcin is con- #P194 POSTER SESSION IV: CHEMICAL
sistent with the anomalous migration on SDS-PAGE and SIGNALING AND BEHAVIOR; ANIMAL
a tendency to undercharge in electrospray ionisation mass BEHAVIOR/PSYCHOPHYSICS;
spectrometers. All this biophysical and ligand binding data
point to darcin adopting a more stable and compact con-
formation with a smaller ligand binding cavity than related
MUPs. Acknowledgements: These studies were supported COMMUNICATION
by the Biotechnology and Biological Science Research Kirrel-3 is Required for the Coalescence of Vomeronasal
Council (BB/J002631/1). Sensory Neuron Axons into Glomeruli and for Male-Male
Aggression
Jean-François Cloutier1,2, Alexandra Brignall1,2, Tyler

#P193 POSTER SESSION IV: CHEMICAL Cutforth3, Kang Shen4, Janet Prince1,2
Montreal Neurological Institute Montréal, QC, Canada,
BEHAVIOR/PSYCHOPHYSICS; 2
McGill University Montréal, QC, Canada, 3University of
CHEMOSENSATION AND METABOLISM; California Irvine, CA, USA, 4Stanford University Stanford, CA,
VOMERONSASAL AND CHEMICAL USA
COMMUNICATION
The accessory olfactory system controls social and sex-
HCN Channels Mediate Proton-dependent Signaling in the ual interactions in mice that are critical for survival.
Mouse Vomeronasal Organ Vomeronasal sensory neurons (VSNs) form synapses with
Annika Cichy, Tobias Ackels, Jennifer Spehr, Marc Spehr dendrites of second order neurons in glomeruli of the acces-
sory olfactory bulb (AOB). Axons of VSNs expressing the
RWTH Aachen University, Dept. Chemosensation Aachen,
same vomeronasal receptor (VR) coalesce into multiple
Germany
glomeruli within spatially conserved regions of the AOB.
The mouse vomeronasal organ (VNO) plays an important Here we examine the role of the Kirrel family of transmem-
role in the detection of semiochemicals and other social brane proteins in the coalescence of VSN axons within the
cues. However, many of the basic mechanisms that con- AOB. We find that Kirrel-2 and Kirrel-3 are differentially
trol VNO physiology remain largely unknown. Here, we expressed in subpopulations of VSNs and that their expres-
investigate proton-mediated activity in the mouse VNO. sion is regulated by activity. While Kirrel-3 expression is
We show that mouse urine is not only a rich source of not required for early axonal guidance events, such as fas-
social chemosignals, but can also create an acidic environ- ciculation of the vomeronasal tract and segregation of apical
ment for such cues. For females, in particular, we find an and basal VSN axons in the AOB, it is necessary for proper
experience-dependent variation of their generally low uri- coalescence of axons into glomeruli. Ablation of Kirrel-3
nary pH. Using whole-cell patch-clamp recordings from expression results in disorganization of the glomerular layer
visually identified sensory neurons in acute tissue slices of the posterior AOB and formation of fewer, larger glo-
of the mouse VNO, we show that vomeronasal sensory meruli. Furthermore, altered glomerular formation is asso-
neurons are activated by protons. We describe that acidic ciated with loss of male-male aggression in kirrel-3−/− mice.
solutions dose-dependently induce inward currents in volt- Taken together our results indicate that differential expres-
age-clamp measurements and elicit robust action potential sion of Kirrels on vomeronasal axons generates a molecular
firing in current- clamp recordings. Surprisingly, our inves- code that dictates their proper coalescence into glomeruli
tigations suggest no substantial involvement of ‘classical’ within the AOB. Acknowledgements: Canadian Institutes
candidate proton- activated ion channels. Instead, the of Health Research and Natural Sciences and Engineering
pharmacological profile and biophysical properties of the Research Council of Canada.
#P195 POSTER SESSION IV: CHEMICAL Florida State University/Program in Neuroscience, Dept. Biol.
SIGNALING AND BEHAVIOR; ANIMAL Sci. Tallahassee, FL, USA
BEHAVIOR/PSYCHOPHYSICS; Medial amygdala responds differentially to conspecific and
CHEMOSENSATION AND METABOLISM; heterospecific chemosensory signals with different meanings
VOMERONSASAL AND CHEMICAL and different behavioral responses; and it may be responsible
COMMUNICATION for routing information to hypothalamic/preoptic circuits
involved in producing the appropriate responses. The vome-
Transduction for pheromones in the main olfactory
ronasal organ (VNO) is the primary but not only source of
epithelium is mediated by the Ca2+- activated
chemosensory input to medial amygdala but VNO-lesions
channel TRPM5
disrupt the characteristic patterns of responses. The circuit
Diego Restrepo1, Fabian Lopez2, Roberto Lopez1, Juan for processing VNO-driven chemosensory input includes the
Bacigalupo2 main intercalated nucleus (mICN), one of several GABA-ir
ICN cell groups in the amygdala. mICN appears to regulate
1
Department of Cell and Developmental Biology, posterior medial amygdala (MeP) activity similarly to the
Neuroscience Program and Rocky Mountain Taste and Smell regulation of central and basolateral amygdala activity by
Center, University of Colorado Anschutz Medical Campus paracapsular ICN cell groups, in the fear conditioning cir-
Aurora, CO, USA, 2Department of Biology, University of cuit. Using immediate-early gene expression, we previously
Chile Santiago, Chile found that GABA-receptor-ir cells in MeP are suppressed by
The main olfactory epithelium contains olfactory sensory heterospecific stimuli –as mICN GABA-ir cells are activated.
neurons (OSNs) that respond to odorants. Interestingly, Now using brain slice recording we show hyperpolarization
there is growing evidence that some OSNs in this epithelium of MeP cells by field stimulation of local mICN. Preliminary
respond to pheromones through an unknown transduction evidence also suggests mICN cells are suppressed by bath-
mechanism. Here we report on a survey for pheromone applied dopamine, as is the case for paracapsular ICN cells.
transduction in a subset of OSNs expressing the transient The amygdala contributes to the motivational/emotional
receptor potential channel M5 (TRPM5), a Ca2+ -activated evaluation of sensory inputs of all modalities and for diverse
monovalent cation-selective channel. As in the majority of behaviors. This concordance between amygdala processing
OSNs, the cyclic nucleotide-gated (CNG) channel subu- in completely different types of behavior may indicate some
nit A2 is expressed in the cilia of OSNs expressing GFP commonalities in circuit organization. Dopamine may be
under control of the TRPM5 promoter. Interestingly these part of a mechanism for modulating amygdala processing
TRPM5-GFP+ OSNs lack the Ca2+-activated Cl- channel of sensory information according to brain state or previ-
ANO2 found in the majority of OSNs. Complementary ous experience. Dopamine appears to modulate experience-
loose patch current and Ca2+ fluorescence recordings show dependent chemosensory responses in basolateral amygdala
that TRPM5- GFP+ OSNs respond to pheromones and not but so far we have not demonstrated an effect in medial
to odorants, while TRPM5-GFP- OSNs respond to both. amygdala. Acknowledgements: Supported by NIDCD
Finally complementary pharmacological and TRPM5 grants R01-DC005813, T32-DC000044 and funding from
knockout experiments show that TRPM5-GFP OSNs Florida State University
respond to pheromones through the TRPM5 channel. Thus,
pheromone responses of TRPM5-GFP+ OSNs are mediated #P197 POSTER SESSION IV: CHEMICAL
by ciliary Ca2+ influx through CNG that gates opening of SIGNALING AND BEHAVIOR; ANIMAL
TRPM5. Acknowledgements: Funded by NIH DC006070 BEHAVIOR/PSYCHOPHYSICS;
and DC004657 (D.R.), CONDECYT 1100682 (F.L.) and
CONICYT and MECESUP UCH0713 (J.B.)
COMMUNICATION
#P196 POSTER SESSION IV: CHEMICAL Interspecies communication mediated by tear fluids
SIGNALING AND BEHAVIOR; ANIMAL Mai Tsunoda, Kazushige Touhara
Department of Applied Biological chemistry, The University
of Tokyo Tokyo, Japan
COMMUNICATION Communication between animals are regulated by a vari-
ety of chemical cues emitted from the body fluids. Recent
AMYGDALAR PROCESSING OF SALIENT CHEMOSENSORY
works have revealed that exocrine grand-secreting peptide
SIGNALS
1 (ESP1), which is released from male mouse tear fluids,
Lindsey M Biggs, Ariel R Simonton, Michael Meredith enhances female sexual behavior through the vomeronasal
organ. This data indicates that the tear fluid is one of the steroids, a class of chemicals that originally isolated from
important sources of chemical cues. However, it is unknown mouse urine, we classified a total of 20, 853 responsive
whether tear-derived chemical cues mediate only intraspe- neurons into 17 functional types. We found that the large
cies communication. In this study, we aimed to understand majority of functional receptor types present in equal
a novel function of tear fluids in interspecies communica- abundance in males and females. However, we found clear
tion by focusing on tear fluids of rats, a predator of mice. sexual dimorphism, as two functional types appeared to be
First, we examined the effect of rat tear fluids on the mouse male specific, including an epitestosterone- selective recep-
vomeronasal system. c-Fos analysis revealed that rat tear tor type 100-fold more abundant in males than in females.
fluids contained some stimulants that induced c-Fos expres- To explore the mechanism generating this dimorphism, we
sion in the accessory olfactory bulb (AOB), the first center found male specific receptor types became rare after long-
of the vomeronasal system. It has been known that rats have term exposure to the odors of female mice, with the result
10 members of ESP family genes, therefore, we examined that the vomeronasal organs from males were converted
whether the stimulants in rat tear fluids are ratESPs. Western to a pattern indistinguishable from females. This differ-
blot analysis indicated that ratESP5 and ratESP7 were ence in AOS receptor type is by far the strongest sexual
secreted in rat tear fluids. However, recombinant ratESP5 dimorphism ever reported in the mammalian central nerv-
and ratESP7 did not induce c-Fos expression in the mouse ous system; that this dimorphism is determined entirely
AOB. This data suggests that there exists novel mouse vome- by experience indicates that a sensory system devoted to
ronasal stimulants in rat tear fluids. We next purified the “innate” responses is strongly modulated by rearing con-
stimulants from rat tear fluids by activity-based fractiona- ditions. Acknowledgements: This study was funded by
tion. Amino-terminal peptide sequence and genome analy- NIH- NINDS/NIAAA Grant R01 NS068409, and NIH
sis revealed that a c-Fos-inducing peptide was encoded by a Director’s Pioneer Award DP1 OD006437(T.E.H.)
gene whose function has not been revealed. We named this
peptide P18. Recombinent P18 induced c-Fos expression in
the AOB of wild type mice, but not in the TRPC2 knock- #P199 POSTER SESSION IV: CHEMICAL
out mice. These results suggest a possibility that P18 in rat SIGNALING AND BEHAVIOR; ANIMAL
tear fluids mediate interspecies communication through the BEHAVIOR/PSYCHOPHYSICS;
vomeronasal organ. CHEMOSENSATION AND METABOLISM;
COMMUNICATION
SIGNALING AND BEHAVIOR; ANIMAL Palatability of Cycloheximide or Caffeine Mixed with Sugar
BEHAVIOR/PSYCHOPHYSICS; in Hamsters
CHEMOSENSATION AND METABOLISM; Elizabeth M. Casey, Bradley K. Formaker, Thomas
VOMERONSASAL AND CHEMICAL P. Hettinger, Marion E. Frank
COMMUNICATION University of CT Health Center Farmington, CT, USA
Experience-dependent plasticity causes sexual dimorphism Multiple coding mechanisms for bitter stimuli are sug-
in mouse pheromone-sensing neurons gested by taste receptive-field specificity, quality and pal-
Pei S. Xu, Timothy E. Holy atability. Quality and palatability of quinine (Qui), salicin
(Sal), caffeine (Caf) and cycloheximide (Cyc) differ in ham-
Department of Anatomy & Neurobiology, Washington
sters (Mesocricetus auratus). Quality, studied with gener-
University School of Medicine St. Louis, MO, USA
alizations of conditioned taste aversions (CTA), differs for
In mice, the normal expression of most sex-specific behav- each; but, each stimulus is aversive measured in preference
iors requires an intact accessary olfactory system (AOS). to water. Hamster taste aversions are generally reduced
While the AOS has been long viewed as a sexually dimor- by adding sucrose. Thus, aversions to 1mM Qui, 10mM
phic circuit, the known anatomical differences between Sal, 30mM Caf and 30µM Cyc were compared to the 4
males and females consist of modest changes in the pack- stimuli mixed with 500mM sucrose. Qui and Sal palatabil-
ing of neurons in particular brain regions. By themselves, ity increased with sucrose added but Caf and Cyc did not
these differences may be insufficient to explain observed (Lloyd et al. 2012). To determine concentration- depend-
dimorphic behaviors. Here we asked whether the first ence, 5, 10 and 30mM Caf and 5, 10 and 30mM Cyc (with
order neurons, AOS sensory neurons differed functionally and without 500mM sucrose) were tested for 2-bottle 48-hr
between two sexes. Using light-sheet based high-speed cal- preference vs. water (R-L bottle positions reversed daily).
cium imaging technique, we recorded ~260,000 individual Each hamster randomly received 14 stimuli based on a mod-
neurons in intact vomeronasal epithelia from male and ified Latin Square. A preference ratio [ml stimulus ingested/
female mice. According to the cell responses to 12 sulfated ml total fluid ingested; indifference = 0.5] was computed
and differences tested with analysis of variance (a =0.05). in TNF-deficient mice. Together, these results suggest that
Hamsters strongly preferred sucrose over water [0.725]. Caf TNF may be an important mediator for taste dysfunction
and Cyc aversions were unaffected by concentration or the associated with inflammation. Acknowledgements: This
addition of sucrose. Average preference ratios collapsed study was supported by NIH/NIDCD grants DC010012
across concentration were 0.210 for Caf and 0.225 for and DC011735.
Caf + sucrose; 0.164 for Cyc and 0.151 for Cyc + sucrose.
Reducing bitter stimulus concentration did not increase
amelioration by sucrose. This is consistent with Cyc aver- #P201 POSTER SESSION IV: CHEMICAL
sions (1-hr tests) quickly developing, even to Cyc-sucrose SIGNALING AND BEHAVIOR; ANIMAL
mixtures without affecting sucrose intake (Hettinger et al. BEHAVIOR/PSYCHOPHYSICS;
2007), and Cyc serving as a CTA UCS when injected IP CHEMOSENSATION AND METABOLISM;
(Formaker et al. 2009). Acknowledgements: Supported by VOMERONSASAL AND CHEMICAL
UConn SDM Alumni Research Fellowship and NIH grant COMMUNICATION
DC004099.
Evidence of neonatal memory of odor configuration
Gérard Coureaud1, Thierry Thomas-Danguin1, Donald
#P200 POSTER SESSION IV: CHEMICAL A. Wilson2, Guillaume Ferreira3
CSGA - Centre des Sciences du Goût et de l’Alimentation
/ Developmental Ethology and Cognitive Psychology Team
CHEMOSENSATION AND METABOLISM; & Flavour Perception Team Dijon, France, 2Emotional Brain
VOMERONSASAL AND CHEMICAL Institute / New York University School of Medicine New
COMMUNICATION York, NY, USA, 3Nutrition and Integrative Neurobiology
Regulation of Taste Responses by TNF (NutriNeuro) Group / INRA 1286, Université de Bordeaux
Bordeaux, France
Jinghua Chai, Pu Feng, Hong Wang
The perception of some mixtures of odorants engages
configural abilities, i.e. the perception of these mixtures
Patients with inflammatory diseases often experience taste as single odor objects. For instance, data in human adults
alterations. Yet, how inflammation affects taste function demonstrated that a mixture of two odorants (AB), one
is not fully understood. Previously, we showed that tumor smelling like strawberry and the other like caramel, gen-
necrosis factor (TNF), a potent proinflammatory cytokine, erates the configural perception of the odor of pineap-
is preferentially expressed in a subset of type II taste cells. ple (Le Berre et al., 2008; Barkat et al., 2012). Configural
The level of TNF in taste cells can be further induced by processing may be adaptive also for young organisms, to
inflammatory stimuli such as lipopolysaccharide (LPS), a which rapid extraction of chemical information from the
bacterial cell-wall component that elicits acute inflamma- maternal environment, highly complex, is a prerequisite
tion. Although TNF plays important roles in mediating to survival. Thus, results in newborn rabbits suggest the
inflammation and cell death in various tissues, its roles in perception of a unique odor in the AB mixture (smelling
taste buds remain to be determined. In this study, we car- like configural pineapple in humans) and different from
ried out taste behavioral tests and gene expression analy- the odors of the elements (Coureaud et al., 2008, 2009a).
ses in wild-type and TNF- deficient mice. Lickometer tests To clearly demonstrate that the configural AB perception
were conducted to examine behavioral responses to salty, does not directly depend on A and B perception, we inves-
sour, bitter, sweet, and umami taste compounds before and tigated here whether rabbit neonates recognize the AB mix-
after LPS-induced inflammation. Our results showed that ture even in the absence of A and B recognition. To that
TNF-deficient mice were less sensitive to the bitter com- goal, rabbit pups were conditioned to AB on day 1. On
pound quinine before any treatments. After LPS injection, day 2, recall of A and recall of B were followed by intra-
wild-type mice displayed a range of altered responses to peritoneal injection of either saline or a pharmacological
the taste compounds, especially to the sweet taste com- amnesic agent (see Coureaud et al., 2009b, 2011). Testing
pound sucrose. In contrast, TNF-deficient mice did not for behavioral responsiveness to A, B and AB occurred on
show a significant alteration in response to sucrose after day 3. Control pups responded behaviorally to AB but also
LPS treatment, suggesting that TNF plays an important to A and B. As expected, the pups injected with the amne-
role in regulating taste response to sucrose under LPS- sic agent did not respond to A and to B. However, they
induced inflammation. Furthermore, gene expression responded to AB, indicating an AB perception independent
analyses by quantitative RT-PCR showed that the levels of A and B representations. In summary, the present results
of several inflammation- and cell-death- related genes were confirm the perception by rabbit neonates of a configura-
increased by LPS in wild-type mice, but were not induced tion in the AB mixture, and demonstrate for the first time
the neonatal ability to memorize odor mixtures as con- by R01 DC009613 and based upon work supported
figurations independent of the memory of their elements. by the U. S. Army Research Office under grant number
Acknowledgements: Supported by French ANR-2010- W911NF-11-1-0087. Sensor implantation was performed
JCJC-1410–1 MEMOLAP to GC, TTD and GF. at The Behavioral and Physiological Phenotyping Core,
which is supported, in part, by funding from the NIH-
NIDCD Core Grant 1P30DC011735-02.
BEHAVIOR/PSYCHOPHYSICS; #P203 POSTER SESSION IV: CHEMICAL
CHEMOSENSATION AND METABOLISM; SIGNALING AND BEHAVIOR; ANIMAL
VOMERONSASAL AND CHEMICAL BEHAVIOR/PSYCHOPHYSICS;
COMMUNICATION CHEMOSENSATION AND METABOLISM;
Sniffing Strategies in Wild-Type and Olfactory Marker
Protein Knock-Out Mice
COMMUNICATION
The Temporal Structure of Odor Mixture Perception in Rats
Glen J. Golden1, Johannes Reisert1, Alan Gelperin1,2
Leslie M Kay1,2, Nisarg M Mehta1, Cinar Doruk3
1
2
Princeton Neuroscience Institute, Department of Molecular 1
Institute for Mind and Biology, University of Chicago
Biology, Princeton University Princeton, NJ, USA Chicago, IL, USA, 2Department of Psychology, University of
Chicago Chicago, IL, USA, 3St. John’s College Annapolis, MD,
Detection and identification of an odor requires nasal
USA
inhalation or sniffing behavior that delivers odorants to
the olfactory receptor neurons (ORNs) deep in the nasal Temporal structure of odor mixtures is often overlooked,
cavity. Our experiments combine behavioral assessment but this is what gives many mixtures their complex per-
of odor detection and discrimination tasks with meas- cepts. This structure can help us understand the puzzling
urements of sniffing behavior to clarify the strategies a qualities of odor mixtures. There is not yet any theory that
mouse uses when confronted with odor-based learning can predict whether a mixture of 2 monomolecular odor-
tasks and the mechanisms underlying odor perception. ants will smell like both odors (elemental), like one more
We study the behavior and sniffing patterns of mice with than the other (overshadowing), or like neither (configu-
(WT) or without (KO) functional olfactory marker pro- ral or synthetic). One feature that is often overlooked in
tein (OMP), a protein that is responsible for speeding studies of mixture perception is the difference in percep-
up the time course of odor-induced responses in ORNs. tual arrival time for the two odors. These delays have been
OMP KO and WT mice were implanted with wireless measured in humans, but implementing these sensitive
pleural pressure sensors to record sniffing patterns. These perceptual assays in rats is much more difficult. We have
mice were then trained and tested in go/no go odor dis- developed a task that allows us to do this in rats, using a
crimination tasks to distinguish solvent (mineral oil) odor combined 2-alternative choice - go/no-go paradigm. The
from the odor of 1-propanol 10−4 log dilution. Upon pro- results show that behavioral response profiles to timing
panol or solvent exposure, WT mice increased their sniff- differences in binary mixtures are specific to the odor pair
ing rate from ~4 Hz to 10 Hz and maintained a higher and that the responses to positive and negative delays are
sniffing rate for rewarded (S+) trials. However, KO mice not symmetrical. For example, at zero delay (only the natu-
continued to increase their sniff rate following the onset ral processes producing delays) in a 1:1 mixture of amyl
of the odor (S+) and solvent-odor cues (S−) significantly acetate and anisole, anisole overshadows amyl acetate. As
in comparison to the WT mice (p <0.05). This suggests anisole is moved earlier in time overshadowing becomes
that OMP KO mice require more stimulation cycles to stronger. In the negative direction, with amyl acetate pre-
determine the identity of an odor cue in contrast to OMP ceding anisole, the mixture enters a configural regime at
WT mice, which make a determination quickly and begin −50ms to −200ms and then takes on an elemental qual-
to decrease sniff frequency immediately following odor ity at −250ms. These results suggest that in the temporal
onset. Understanding the temporal relationship of both domain, elemental and configural responses are close,
sniffing frequency and the duration of odor sampling in with overshadowing responses occupying a separate part
OMP WT and KO mice, in combination with biophysical of the temporal space. The properties of odorants, such
data on odor responses obtained from individual ORNs as sorptiveness and volatility, that may contribute to tem-
isolated from these mice, will help elucidate the role of poral effects are discussed. Acknowledgements: Institute
OMP in olfactory transduction and odor-investigation for Mind and Biology Seed Grant (LK) Hodson Research
strategies. Acknowledgements: This research is supported Fellowship (CD)
#P204 POSTER SESSION IV: CHEMICAL Sébastien Romagny, Thierry Thomas-Danguin, Gérard
SIGNALING AND BEHAVIOR; ANIMAL Coureaud
BEHAVIOR/PSYCHOPHYSICS; CHEMOSENSATION Centre des Sciences du Goût et de l’Alimentation (CSGA),
AND METABOLISM; VOMERONSASAL AND UMR 6265 CNRS, UMR 1324 INRA, Université de Bourgogne,
CHEMICAL COMMUNICATION Flavour Perception Team and Developmental Ethology and
The Fine Temporal Structure of the Rat Licking Pattern: Cognitive Psychology Team Dijon, France
What Causes the Variability in the Interlick Intervals and Comparative studies give the opportunity to better under-
How is it Affected by the Drinking Solution? stand common and dissimilar processes in terms of percep-
Xiong B. Lin1, Dwight R. Pierce2, Kim E. Light2, Abdallah tion, cognition and behavior between organisms. Regarding
M. Hayar1 the elemental and configural perception of odor mixtures,
1
University of Arkansas for Medical Sciences, Dept. of newborn rabbits and human adults present some similari-
Neurobiology & Developmental Sciences Little Rock, AR, ties. In particular, they both process a 6-component mixture
USA, 2University of Arkansas for Medical Sciences, Dept. of (RC) in a weak configural way, leading to the perception of
Pharmaceutical Sciences Little Rock, AR, USA a novel odor (e.g., red cordial in humans) in the mixture:
in rabbits, neonates do not respond to RC after learning of
Licking is a repetitive behavior controlled by a central pattern one component, while they generalize for another mixture
generator. Even though interlick intervals (ILI) within bursts of same complexity; in humans, the quality of the single
of licks are considered fairly regular, the conditions that affect odorants is judged as significantly different to the mixture.
their variability are unknown. We analyzed the licking pat- Here, we set out to examine whether the perception of the
tern in rats that were licking water, 10% sucrose solution, or RC configuration strictly depends on the quantity and/or
10% ethanol solution, in 90 min recording sessions after 4 h the quality of the components. To that goal, we carried out
of water deprivation. The histograms of ILIs indicate that a generalization experiment in rabbit pups and a similar-
licking typically occurred at a preferred ILI of about ~135 ms ity rating task in human adults, using several sub- mix-
with evidence of bi- or multi-modal distributions due to occa- tures of increasing complexity (i.e., 2, 3, 4 or 5 odorants).
sional licking failures. The longer the pause between bursts We conditioned the pups to sub-mixtures and tested their
of licks (≥3 consecutive licks with ILIs <200 ms), the shorter behavioral responsiveness to these stimuli compared to the
was the first ILI of the burst. When bursts of licks were pre- full mixture (RC). In humans, participants rated the simi-
ceded by a pause >4 sec, the ILI was the shortest (~110 ms) at larity between the sub-mixtures and RC. The results indi-
the beginning of the burst and then it increased rapidly in the cated that newborn rabbits became able to respond to the
first few licks and slowly in subsequent licks. The first ILI of a RC mixture when they had previously acquired at least 4
burst of licks was not significantly different when licking any of its components, whatever their odor quality. In human
of the 3 solutions, but subsequent licks exhibited a temporal adults, similarity between sub-mixtures and the RC mix-
pattern characteristic of each solution. Moreover, rats licked ture depended more on the odor quality of the odorants
the ethanol solution in shorter bursts and the sucrose solution included in sub-mixtures rather than on the number of
in longer bursts when compared to water. Therefore, rats may mixed odorants. Therefore, even if these two models shared
rapidly identify the fluid and modify their licking behavior a configural perception of the same RC odor mixture, the
by adjusting the temporal pattern of licks and the number of factors underpinning its perception seem to be different, at
licks/burst. The rapid deceleration in licking rate in bursts of least in part.
licks that occurred after >4 sec pause was due to an increase
from ~27 ms to ~56 ms in the tongue-spout contact duration
while the intercontact interval was only slightly changed (80–
90 ms). Therefore, the contact duration seems to be the major
factor that increases the variability in the ILIs, and could
be another means for the rat to adjust the amount of fluid BEHAVIOR/PSYCHOPHYSICS;
ingested. Acknowledgements: Grant P20 GM103425-09 CHEMOSENSATION AND METABOLISM;
COMMUNICATION
The Contribution of the T1R1 Subunit to Taste Detection of
SIGNALING AND BEHAVIOR; ANIMAL Glutamate as Behaviorally Assessed in a Murine Model.
BEHAVIOR/PSYCHOPHYSICS; CHEMOSENSATION
AND METABOLISM; VOMERONSASAL AND Kimberly R Smith, Alan C Spector
CHEMICAL COMMUNICATION Florida State University Tallahassee, FL, USA
How do rabbit newborns and human adults perceive the Whether taste detection of L-glutamate is mediated solely
configuration in a 6-component blending odor mixture? by the T1R1+T1R3 heterodimer or whether an additional
glutamate-sensing taste transduction mechanism(s) con- size (g/meal, p<0.01), a measure of postingestive feedback.
tributes is controversial. Here, we behaviorally assessed the Total intake and meal size recovered to base line levels
necessity of the T1R1 subunit to taste detection of mono- during the TA treatment. These increases in meal size and
sodium glutamate (MSG) in a two-response discrimination total intake were significantly correlated with increases in
procedure using T1R1 knockout (KO) mice and their same- the expression of PRPs (p’s≤0.02), which were present in
sex littermate wild-type (WT) controls. Water-restricted mice saliva by the third day. TA treatment also decreased rate
were trained to discriminate a tastant from water with a cor- of feeding (g consumed/sec, p<0.01), a measure of palat-
rect response resulting in the delivery of a water reinforcer ability. Although there was an increase in rate of feeding
and an incorrect response resulting in a time-out. Sensitivity during the treatment, which was significantly correlated
to NaCl and MSG was similar between genotypes. However, with PRP production (p=0.03), this measure did not
upon the addition of the sodium channel blocker amiloride return to base line levels. To examine palatability in the
(100 µM) and inosine 5′ monophosphate (IMP), at a concen- absence of postingestive feedback, rats were given TA at
tration (2.5 µM) shown to potentiate the glutamate signal in varying concentrations (0–3%) in a brief-access test while
a variety of assays, performance of the KO mice to this MSG PRPs were either increased (by tannin exposure, n=6) or
mixture (M+A+I) was severely impaired. Whereas WT mice unchanged (maintained on a control diet, n=6) in a sec-
performed at consistently high levels across concentrations, ond experiment. Rats with PRPs present in the saliva licked
the ability of the KO mice to detect the M+A+I solution was more, in 10s, to intermediate concentrations of TA (0.09
above chance only at the higher MSG concentrations. The and 0.18%, p’s<0.04) than rats without PRPs. Together,
possibility that IMP was precluding concentration-depend- these data suggest that PRPs play a role in altering both
ent performance in the WT mice to the MSG in the presence the palatability and postingestive feedback associated with
of amiloride was confirmed when we found that WT mice TA. Acknowledgements: NIH-DC-000044, DK-73936,
could detect 2.5 µM IMP alone with relatively high accu- DC-004785, DC-012632
racy, whereas KO mice could not respond significantly above
chance. Collectively, these results strongly suggest that 1) the
Na+ ion dominates the taste detection of MSG in mice con- #P208 POSTER SESSION IV: CHEMICAL
sistent with other recent data from our laboratory, and 2) glu- SIGNALING AND BEHAVIOR; ANIMAL
tamate may be activating a T1R1-independent high- threshold BEHAVIOR/PSYCHOPHYSICS;
receptor in the presence of IMP, but normal detection of glu- CHEMOSENSATION AND METABOLISM;
tamate depends on the T1R1 subunit. Acknowledgements: VOMERONSASAL AND CHEMICAL
NIH R01-DC004574 (ACS) & NSF GRF to KRS COMMUNICATION
Adaptive respiratory behavior communicates social
#P207 POSTER SESSION IV: CHEMICAL hierarchy in rats
SIGNALING AND BEHAVIOR; ANIMAL Daniel W. Wesson
Case Western Reserve University, Dept. of Neurosciences
CHEMOSENSATION AND METABOLISM; Cleveland, OH, USA
COMMUNICATION Sniffing behavior is regularly displayed by terrestrial verte-
brates during social interactions. No measures of sniffing
A Role for Salivary Proteins in Taste Mediated Behavior from interacting animals are available, however, calling into
Ann-Marie Torregrossa, Michelle B. Bales, Robert question the utility of this behavior within the social context.
J. Contreras, James C. Smith, Lisa A. Eckel Here I found that investigation by a dominant rat towards
the facial region of a subordinate often elicits a concomi-
Dept. of Psychology, Florida State University Tallahassee,
tant decrease in respiratory frequency in the subordinate
FL, USA
animal. Failure of subordinates to decrease their respiratory
While few studies have examined the influence of salivary frequency in this context shortened the latency for agonis-
proteins on taste-mediated behavior, it has been demon- tic behavior by dominant rats, reflecting that decreases in
strated that salivary proline-rich proteins (PRPs) bind to respiratory frequency serve as appeasement signals. Rats
bitter-tasting tannic acid (TA) and may function to increase rendered unable to smell displayed reciprocal respiratory
the acceptability of TA-containing diets. To test this behavior, demonstrating the independence of this behavior
hypothesis, we collected saliva samples and measured spon- for gathering odors or sharing odor stimulus space. Oxytocin
taneous feeding behavior in rats (n=8) fed a control diet treatment abolished agonistic behaviors and reciprocal res-
(16 days) followed by a diet containing 3% TA (12 days). piratory displays. These findings demonstrate a novel form
Total intake was reduced during the first 3 days of the TA of communication in rodents, by showing that rodents uti-
diet (p<0.01). This reduction was due to a decrease in meal lize sniffing behaviors communicatively, not only to collect,
but also to convey information. Acknowledgements: NSF #P210 POSTER SESSION IV: CHEMICAL
grant IOS- 1121471 SIGNALING AND BEHAVIOR; ANIMAL
#P209 POSTER SESSION IV: CHEMICAL CHEMOSENSATION AND METABOLISM;
SIGNALING AND BEHAVIOR; ANIMAL VOMERONSASAL AND CHEMICAL
BEHAVIOR/PSYCHOPHYSICS; COMMUNICATION
CHEMOSENSATION AND METABOLISM; The Gustatory Stop-Signal Task: A Method for Measuring
VOMERONSASAL AND CHEMICAL Taste Quality Discrimination in Mice with Millisecond
COMMUNICATION Temporal Resolution
Discrimination of homologous alcohol odorants varying in Dustin M Graham, David L Hill

carbon chain by behaviorally-trained Fisher F344 rats UVA/Psychology Charlottesville, VA, USA
Wendy M Yoder1, 2, Jennifer L Bizon2,4, Barry Setlow2,4,5, There is a lack of consensus regarding the roles of tem-
David W Smith1, 2, 3 poral and spatial coding of taste quality in the gustatory
1
Department of Psychology, Behavioral Neuroscience, system due in part to various experimental and analytical
University of Florida Gainesville, FL, USA, 2UF Center for differences among previous studies. Quantitative behav-
Smell and Taste Gainesville, FL, USA, 3Department of ioral analysis can be used to test for the cognitive prin-
Otolaryngology - Head and Neck Surgery, University of ciple of speed-accuracy tradeoff (SAT), a hallmark of
Florida Gainesville, FL, USA, 4Department of Neuroscience, temporal processing of sensory stimuli. However, current
College of Medicine, University of Florida Gainesville, methods used to study taste perception in rodents are
FL, USA, 5Department of Psychiatry, University of Florida temporally too slow for precise reaction-time measure-
Gainesville, FL, USA ments required to test for SAT in the gustatory system.
We designed a novel behavioral paradigm, the Gustatory
Recent studies suggest that chemicals varying in car- Stop- Signal Task, in head-restrained mice for measur-
bon chain length may represent a generalization gradi- ing perceptual identification of taste stimuli with mil-
ent among odorants. To investigate how odor quality is lisecond temporal resolution. Using this new paradigm,
reflected perceptually in the rat, we measured graded per- we will apply threshold psychophysics to determine if a
ceptual similarity by varying carbon chain length across stimulus-dependent SAT is present during discrimina-
a series of homologous alcohol pairs. We employed an tion of basic tastes. This will provide crucial behavioral
automated, liquid-dilution olfactometer to train F344 rats evidence for the potential roles of temporal and spatial
on a two-odor discrimination task. Six odorants (1-pro- coding strategies underlying taste quality coding in the
panol, 1-butanol, 1-pentanol, 1-hexanol, 1-heptanol and gustatory system. Additionally, the task can be combined
1-octanol) were arranged to produce 30 novel odorant pairs with advanced physiological techniques, such as visually
differing between one and five carbon atoms; testing ses- guided whole-cell patch-clamp recordings in sub-regions
sions included presentation of only one randomly assigned of gustatory cortex. The gustatory stop-signal task in
pair daily (200 trials daily). Results showed that, although head-restrained mice will provide a new foundation to
rats learned to discriminate between any two odorant pairs, combine precise quantitative behavioral measurements of
discrimination accuracy changed systematically with car- taste perception alongside state-of- the-art in vivo physi-
bon chain length difference. Error patterns were remark- ology. Acknowledgements: NIH Grants R01 DC00407
ably consistent across animals, such that marked increases and F32 DC012461 - 01A1
in misses and false alarms were indicated for pairs differ-
ing by one or two carbon atoms. Across all odorant pairs,
these effects were most pronounced during the first 20 tri- #P211 POSTER SESSION IV: CHEMICAL
als. Notably, the greatest degree of perceptual confusion SIGNALING AND BEHAVIOR; ANIMAL
was displayed for two pairs differing by a single carbon BEHAVIOR/PSYCHOPHYSICS;
atom, 1-propanol/1-butanol and 1-heptanol/1- hexanol. CHEMOSENSATION AND METABOLISM;
These data provide further support for carbon chain length VOMERONSASAL AND CHEMICAL
as an important odorant stimulus dimension for study of COMMUNICATION
olfactory receptor interaction (Johnson and Leon, 2000) as
well as demonstrate how hierarchical chemotopic organi- Free Access to Highly Palatable Food during Adolescence
zation of the olfactory bulb may be reflected perceptually. Increases Anxiety- and Depression-like Behaviors in Males,
Furthermore, development of an animal model using the but not in Females
carbon chain paradigm may be useful for assessing the Jeong-Won Jahng, Jin-Young Kim, Joo-Young Lee,
mechanisms underlying olfactory dysfunction. Jong-Ho Lee
Seoul National University School of Dentistry Seoul, that has been borne out in behavioral data. It remains pos-
South Korea sible, however, that a more rapidly developing AN might
We have reported that a long-term access to highly palatable escape detection in time-averaged accounts of behavior
food (HPF) modulates the hypothalamic-pituitary-adrenal such as consumption. With this in mind, we performed a
(HPA) axis response to restraint stress in adult male rats. comparison of AN in two contexts, a two-bottle test and
Psycho-emotional disorders frequently involve dysfunctions a brief access test (which allowed a real-time analysis of
in the HPA axis activity. In this study, male and female SD licking microstructure). At the level of overall consump-
rats had free choices of chocolate cookies as HPF and chow tion, data from the two tasks were in good accord—both
with ad libitum access from PND 28, and then were sub- revealed AN to 28mM saccharin but not to 2.8mM saccha-
jected to behavioral tests at youth. Control group received rin. Additionally, however, the brief access task revealed
chow only, and food conditions were continued through- an initial hesitation to consume the higher concentration
out the whole experimental period. Body weight gain and saccharin solution; this seemingly neophobia-related hesi-
daily caloric intake did not differ between HPF and control tation not only decreased between sessions 1 and 2, but
groups both in males and females. Total ambulatory activ- also decreased linearly across the twenty minutes of ses-
ity was decreased with HPF access in females, but not in sion; that is, AN began within minutes of the rats’ first
males. However, HPF increased anxiety related behaviors exposure to the taste. These data validate the brief-access
in males; i.e. increased rostral grooming and decreased the task as an paradigm with which to measure AN, and also
open arms stay during elevated plus maze test, but did not reveal aspects of AN—perhaps related to short-term
affect those indexes in females. Immobility duration during plasticity—that appear within minutes of the first taste.
forced swim test was significantly increased with HPF access Acknowledgements: National Institutes of Health, World
in males, but the increase was not reached to a statistical sig- of Work Fellowship
nificance in females. Stress-induced corticosterone increase
was shortened with HPF access both in males and females.
Results suggest that adolescence free access to highly palat-
able food may lead to a dysfunction in the HPA axis activity SIGNALING AND BEHAVIOR; ANIMAL
both in males and females; however, its psycho-emotional BEHAVIOR/PSYCHOPHYSICS;
outcome be worse in males. Acknowledgements: Supported CHEMOSENSATION AND METABOLISM;
by MOEST(2009K001269 2010- 0003642) VOMERONSASAL AND CHEMICAL
COMMUNICATION
#P212 POSTER SESSION IV: CHEMICAL Channelrhodopsin Mice use Temporal Information Encoded
SIGNALING AND BEHAVIOR; ANIMAL in the Olfactory Bulb for Odor Sensation.
BEHAVIOR/PSYCHOPHYSICS; Michelle R. Rebello1,2, Thomas S. McTavish2, David
CHEMOSENSATION AND METABOLISM; C. Willhite1,2, Gordon M. Shepherd2, Justus V. Verhagen1,2
VOMERONSASAL AND CHEMICAL 1
John B. Pierce Laboratory New Haven, CT, USA, 2Yale School
COMMUNICATION of Medicine, Dept. of Neurobiology New Haven, CT, USA
Licking Microstructure Reveals Rapid Attenuation of Odor information is represented by spatio-temporal maps
Neophobia in the olfactory bulb (OB). Spatial maps reflect the con-
Kevin J Monk, Benjamin D Rubin, Jennifer Keene, Donald verging axons of olfactory receptor neurons activated by
B Katz odors, onto their respective glomeruli in the OB. The ori-
gins of temporal patterns of glomerular activation are less
well understood, but odorant receptor affinity as well as
Neophobia—the initial hesitation that many animals show odorant sorption kinetics across the olfactory epithelium
to a novel food—is typically measured by comparing con- could underlie temporal parameters such as onset latency
sumption in the first and second sessions of access to the and rise time. Consistent differences in response dynam-
taste; lower consumption in session 1 denotes neophobia, ics across glomeruli have been found for odor- evoked
and higher 2nd- session consumption denotes attenuation responses in the OB. Further, we have shown, using opti-
of neophobia (AN). AN is thought to represent a bona cal imaging that retronasal and orthonasal bulbar reponses
fide example of learning— neural plasticity induced by an differ in response amplitude as well as temporal dynam-
association between the taste and a safe outcome on ses- ics. It is therefore evident that rich temporal information is
sion 1 changes the response to the tastant during session available in the bulbar response. However it is not known
2. Such long-term plasticity processes require time to com- whether these temporally dynamic responses are behav-
plete, and thus AN should only stabilize 90 min or more iorally relevant. Using transgenic mice expressing ChR2
following the first exposure to the tastant—a prediction under the Thy-1 promoter in the mitral cells and a digital
micromirror device to project sniff- triggered light patterns lasted over variable time scales ranging from 1 to 30 min-
onto the dorsal OB we are able to exert tight spatio-tem- utes, indicative of either short-term or long-term poten-
poral control over OB activity patterns. We find that mice tiation. These results suggest that subpopulations of
trained on a go/no-go task are able to discriminate patterns primary afferent synapses can be potentiated throughout
that are spatially identical but differ temporally. By vary- development, and the presynaptic nerve and/ or postsyn-
ing the relative delay among the same regions activated by aptic target specificity for this potentiation will the focus
light patterns we are able to determine the threshold of of future studies. Acknowledgements: T32DC000011,
temporal discrimination. We find that Thy-1 ChR2 but not RO1DC000288
wild-type mice can make temporal discriminations of less
than 30ms. Our optogenetic study confirms that awake,
behaving mice can use temporal information encoded in #P215 POSTER SESSION V: HUMAN TASTE
the bulbar response. This suggests that temporal coding PSYCHOPHYSICS; OLFACTION RECEPTORS;
can contribute to retronasal and orthonasal odor sensa- TASTE DEVELOPMENT
tion. Acknowledgements: This work is supported by NIH/ Ephrin-B/EphB signaling influences the innervation of
NIDCD Grants R01DC009994 and R01DC011286 and fungiform papillae
NIH Institutional Training Grant T15- LM007056 from
the National Library of Medicine. David Collins1, Natalia Hoshino1, Elizabeth M Runge1, Son
Ton1, Omar Diaz1, Jessica Decker1, Mark Henkemeyer2, M
William Rochlin1
#P214 POSTER SESSION V: HUMAN TASTE 1
Loyola U Chicago/Biology Chicago, IL, USA, 2UT
PSYCHOPHYSICS; OLFACTION RECEPTORS; Southwestern/ Developmental Biology Medical Center/
TASTE DEVELOPMENT Dallas, TX, USA
Potentiation of Primary Afferent Innervation in the Rostral Geniculate ganglion axons innervate fungiform taste buds
Nucleus of the Solitary Tract whereas the trigeminal ganglion neurites innervate the adja-
Robert M. Bradley, James A. Corson cent non-taste epithelium. Owing to the proximity of their
terminal fields, non-diffusible repellents are likely to have
University of Michigan/ Biologic and Materials Sciences Ann
a role in preventing incorrect targeting. Eph receptors and
Arbor, MI, USA
ephrins are cell-attached receptor/ligands capable of trigger-
The development of mature primary afferent circuitry ing contact- dependent repulsion or stabilization. An anti-
results from a Hebbian-like synaptic strengthening in a body that detects EphB1, B2, and B3 stains both geniculate
number of sensory systems. This synaptic strengthening and trigeminal axons throughout embryonic development.
is accompanied by a pruning of non-strengthened inputs Anti-ephrin-B1 and anti- ephrin-B2 labeled the dorsal lin-
resulting in a mature terminal field anatomy. Gustatory gual epithelium uniformly at E17, when axons first penetrate
afferents innervating the oral cavity project to the rostral the papilla epithelium in rat. In ephrin-B2 lacZ mice, label
nucleus of the solitary tract (rNTS), the terminal fields was also observed throughout the dorsal epithelium but
of which prune between postnatal days 15 and 60 into an only from E16.5, well after initial invasion of axons into the
adult-like organization. However, it is not known whether epithelium (E14.5). However, in ephrin-B1 lacZ mice, only
any activity dependent changes in synaptic strength can fungiform papilla epithelium was labeled, and this labeling
be induced during this period that may correspond to the was observed at E14.5. Mice lacking EphB1 and EphB2
anatomical remodeling. To investigate the activity-mod- exhibited normal levels of gustatory innervation at E13.5,
ulated plasticity of primary afferent inputs to the rNTS, but less innervation than controls by E17.5, suggesting that
acute horizontal rNTS slices were prepared from rats of ephrin-B/EphB forward signaling may have a stabilizing
postnatal ages covering the period of anatomical plasticity. influence on gustatory afferents in vivo. In vitro, substra-
The solitary tract was stimulated with a concentric bipo- tum stripes prepared from high concentrations of ephrin-
lar electrode and excitatory postsynaptic currents (ePSC) B2-Fc (40 ug/ml) repel neurites from both the geniculate
were recorded. The ability of a synapse to be potentiated and trigeminal ganglia, and this did not depend on which
was investigated by stimulating the solitary tract at 50 Hz neurotrophin was used to promote growth. Stripes pre-
paired with a 5 ms depolarization. Following this tetanic pared from lower concentrations of ephrin-B2-Fc (4 ug/ml)
stimulation increases in ePSC amplitude and rise slope were not repellent; indeed, coverglasses coated uniformly
were observed in each age examined, though only in a with 4 ug/ml ephrin-B2 mildly promoted trigeminal neur-
subset of neurons at each age. Interestingly, the probabil- ite outgrowth length, depending on the stage and neurotro-
ity of inducing potentiation appears to decrease between phin. We are currently analyzing the effects of ephrin-B1
postnatal days 20–25 and subsequently increases from on geniculate and trigeminal neurites. Acknowledgements:
postnatal day 30 onward. Primary afferent potentiation 1R15DC010910-01
#P216 POSTER SESSION V: HUMAN TASTE neurotrophins and the glial cell-line derived neurotrophic
PSYCHOPHYSICS; OLFACTION RECEPTORS; factor (GDNF) family ligands (GFLs), are critical for the
TASTE DEVELOPMENT establishment of proper connections between neurons and
their targets. The four GFLs, GDNF, neurturin (NRTN),
Glial Contributions to the Formation of the Solitary Tract artemin (ARTN) and persephin (PSPN), are potent growth,
and the Rostral Nucleus of the Solitary Tract guidance and survival factors for both PNS and CNS neu-
Sara L Corson, Robert M Bradley, Charlotte M Mistretta rons. The GFLs bind with high affinity to GPI-anchored
coreceptors called the GFRalphas, of which there are four
University of Michigan School of Dentistry Department of
(GFRalpha1-4). In the PNS, these GFL- GFRalpha com-
Biologic and Materials Sciences Ann Arbor, MI, USA
plexes then associate with and activate their common receptor
The solitary tract (ST) consists of afferent fibers that origi- tyrosine kinase, Ret. Several of the GFLs and their receptors
nate in the oral cavity and project to the rostral nucleus of are expressed by components of the peripheral gustatory
the solitary tract (rNST), the site of the first synaptic relay in system. GDNF and NRTN are expressed throughout the lin-
transmitting taste-related information to higher brain areas. gual epithelium, and GDNF, GFRalpha1, GFRalpha2 and
We are interested in the regulatory elements that direct ST Ret are all expressed in taste buds of circumvallate papillae.
formation and rNST development. Glia represent approxi- In addition, neurons of the petrosal and geniculate ganglia
mately half of the cells in the CNS and play roles in neuron express Ret and cognate GFRalphas. It is not known, how-
guidance and synapse development and function. However, ever, whether GFLs function in the development and main-
the time course of glial development and the role of glia tenance of peripheral gustatory circuits. Using conditional
in the gustatory brainstem are unknown. We surveyed the transgenic deletion of Ret, we are in the process of deter-
expression of glial and neuronal markers in the pre- and post- mining whether the GFL/Ret signaling pathway is necessary
natal developing rat ST and rNST to characterize their con- for the development of fungiform papillae, taste buds within
tribution to the development of the gustatory brainstem. We circumvallate and fungiform papillae, and their respective
examined the expression of neuronal markers, including cal- innervation by petrosal and geniculate neurons. These stud-
bindin and NeuN, and glial markers, including glial fibrillary ies will establish whether other neurotrophic factors besides
acidic protein (GFAP), myelin basic protein (MBP) and brain BDNF and NT-4 have developmental functions in the embry-
lipid binding protein (BLBP), in conjunction with P2X2, a onic morphogenic events in the lingual epithelium necessary
marker of gustatory nerve terminal fields, in the developing for papillae and taste bud development, as well as growth
ST and rNST. We found persistent but dynamic expression of and survival functions for the sensory neurons that innervate
calbindin, GFAP and BLBP throughout pre- and post-natal these lingual structures. Acknowledgements: BAP: NINDS
development. In particular, GFAP expression shifts from R01 NS058510 CRD: NIDCR TEAM Tissue Engineering
more fibrillary to more astrocyte-like labeling in the early and Regeneration Training Grant T32 DE007057
postnatal period. This shift is concurrent with terminal field
plasticity, i.e., developmental remodeling and fine-tuning
of circuits. Furthermore, the GFAP-positive astrocyte-like #P218 POSTER SESSION V: HUMAN TASTE
cells are differentially distributed throughout the rNST. This PSYCHOPHYSICS; OLFACTION RECEPTORS;
work highlights potential neuron-glia interactions that are TASTE DEVELOPMENT
important for the development of the gustatory brainstem.
The results provide basic information for building mechanis- The p75 receptor regulates gustatory innervation patterns
tic studies regarding glial function in taste circuit formation. during development
Acknowledgements: NIH NIDCD Grant DC009982 Da Fei, Robin Krimm
University of Louisville/Anatomical Sciences and
Neurobiology Louisville, KY, USA
#P217 POSTER SESSION V: HUMAN TASTE
PSYCHOPHYSICS; OLFACTION RECEPTORS; As a pan receptor of neurotrophins, p75 can function as either
TASTE DEVELOPMENT a pro-survival or pro-death factor during development. P75 is
expressed in both taste buds and taste (geniculate) neurons;
Functions of the GDNF family of neurotrophic factors in the however, the role of p75 receptor during taste development is
development of the peripheral gustatory system unclear. Here we examined the role of p75 in neuron survival,
Christopher R. Donnelly, Brian A. Pierchala taste bud formation and peripheral innervation patterns during
development. We found that p75−/− mice began to lose about
University of Michigan School of Dentistry/Biologic and
22% (p=0.05) of geniculate neurons compared to the wild type
Materials Sciences Ann Arbor, MI, USA
mice at E14.5, and the loss continued to around 36% (p<0.05)
During development of the peripheral nervous system by E18.5. At birth, taste bud number was reduced in p75−/− mice
(PNS), target-derived neurotrophic factors, such as the (60 ± 4, p<0.05) compared to wild type mice (103 ± 7). Remaining
taste buds consisted of two-populations, larger innervated taste buds and in the surrounding stratified epithelium. Although
buds and smaller un-innervated ones. The loss of innervation to mG-positive cell numbers decline by 7 days, induced labeling
taste buds was much greater than would be expected from the is still apparent in taste buds after 2 months. Importantly, we
neuron loss, indicating that innervation could also be disrupted. found no evidence for TM-induced labeling of cells outside
Using DiI-labeling and immunohistochemistry, we found that of taste papilla boundaries. These data indicate that Lgr5
taste nerve growth was delayed and normal innervations pat- expression marks long-term taste cell progenitors, that Lgr5
terns were disrupted in p75−/− mice. Interestingly, the pattern may be a regulator of taste epithelium homeostasis, and that a
of disrupted innervation in p75−/− mice was unique and did duct-basal trench-papilla axis of epithelial renewal may exist.
not resemble that of other neurotrophin knockouts. At E14.5, Acknowledgements: Supported by NSF REU #1062645
which is the first day of target innervation, few axons reached
the tongue epithelium in p75−/− mice. At E16.5, innervation #P220 POSTER SESSION V: HUMAN TASTE
patterns in p75−/− mice were still disrupted, some fiber bundles
PSYCHOPHYSICS; OLFACTION RECEPTORS;
were seen innervating the tip and back of the tongue; however,
innervation to the fungiform papillae in the middle area of the TASTE DEVELOPMENT
tongue was lost in p75−/− mice. This loss of innervation was not Temporal and spatial differences in BDNF and NT4
rescued at a later age, E18.5. Taken together, these results indi- expression determine their unique roles in gustatory
cate that while p75 may mediate neurotrophin support of taste development
neuron survival, it has a unique role in establishing peripheral Tao Huang1, Robin Krimm1
innervations patterns. Acknowledgements: National Institutes
of Health Grant DC009418.
1
University of Louisville/Department of Anatomical
Sciences and Neurobiology Louisville, KY, USA, 2University
of Louisville/ Department of Anatomical Sciences and
#P219 POSTER SESSION V: HUMAN TASTE Neurobiology Louisville, KY, USA
A limited number of growth factors are capable of regulat-
TASTE DEVELOPMENT
ing numerous developmental processes, but how they accom-
Lgr5 Expression Defines a Set of Progenitors that Give plish this is unclear. In the gustatory system, brain-derived
Rise to Both Taste Cells and Keratinocytes of Adult Mouse neurotrophic factor (BDNF) and neurotrophin-4 (NT4) have
Circumvallate and Foliate Papillae different developmental roles but exert their effects through
Theresa A. Harrison1, Anthony M. Downs2, Dennis M. Defoe1 the same receptors (TrkB and p75). Using genome wide
expression analysis, we determined that BDNF and NT4
1
Department of Biomedical Sciences/ETSU College of
regulate the expression of different sets of genes downstream
Medicine Johnson City, TN, USA, 2Warren Wilson College
of receptor signaling in gustatory ganglion. These differ-
Swannanoa, NC, USA
ences in gene expression likely determine their different roles
Lingual taste cells continually turn over in adult mammals; during development. BDNF and NT4 could function differ-
however, the source and properties of regenerative cells ently because of temporal or spatial differences of expres-
remain unknown. Wnt signaling plays critical roles in several sion or theactivation of different signaling pathways. Using
renewing tissues, and the Wnt target Lgr5 has been identified mice in which the coding region for BDNF is replaced with
as a bona fide adult stem cell marker. Because canonical Wnt NT4 (BdnfNt4/ Nt4), we show that NT4 can mediate most of
signaling is activated at sites of mature taste buds, we asked the unique roles of BDNF. Specifically, caspase-3-mediated
if Lgr5 expression identifies analogous stem/progenitor cells cell death, which is increased in Bdnf−/− mice (p<0.01), was
in the taste system. Knock-in mice with the Lgr5-EGFP-ires- rescued in BdnfNt4/Nt4 mice, and the number of caspase-3-mei-
CreERT2 transgene replacing one Lgr5 allele were bred to dated cell death was even lower than that in wild-type mice
the tamoxifen (TM)-inducible mTmG reporter strain. The (p<0.05). In BDNF knockout mice, taste bud innervation in
progeny express soluble GFP (sGFP) under endogenous Lgr5 the tongue was disrupted, and gustatory axons failed to reach
regulation and, following TM treatment, membrane-bound their targets. However, the disrupted innervation was recov-
tdTomato (mT) is replaced by GFP (mG). sGFP is present in ered and targeting is normal when NT4 replaced BDNF. The
all papillae of 7 day-old non-induced mice, but by 12 weeks expression of differentiation-, apoptosis- and axon guidance-
of age is seen only in circumvallate (CV) and foliate papillae. related genes was down-regulated in the geniculate ganglion
Expression in the CV is highest where salivary ducts intersect of BDNF mutant mice (p<0.05), but replacement by NT4
papilla walls, decreases dorsally in the papilla, and is absent rescued all gene expression changes. These findings indicate
from taste buds. In salivary ducts, sGFP is limited to the that the functions of BDNF and NT4 in taste development
outer layer of the bi- layered epithelium. One day after a sin- are interchangeable. Spatial and temporal differences in neu-
gle TM injection, induced mG marks small round cells at the rotrophin expression can regulate differential gene expression
papilla base and, rarely, in the lateral wall of the CV papilla. in vivo and determine their specific roles during development.
By 2–3 days post-injection, labeled cells lie both within taste Acknowledgements: DC009418 and DC007176
#P221 POSTER SESSION V: HUMAN TASTE 2

Department of Dermatology, Medical School, University
PSYCHOPHYSICS; OLFACTION RECEPTORS; of Michigan Ann Arbor, MI, USA, 3Department of Cell
TASTE DEVELOPMENT and Developmental Biology, Medical School, University of
Michigan Ann Arbor, MI, USA
Detailed expression analysis of a neural crest-specific
P0-Cre transgenic mouse line and comparison with Wnt1- Fungiform papillae must contain long-lived sustaining cells
Cre Hongxiang Liu, Yoshihiro Komatsu, Shih-Kai Wang, and short-lived maintaining cells that support development,
differentiation and maintenance of the lateral and apical
Charlotte M Mistretta, Yuji Mishina papilla epithelium and the specialized taste buds. Shh is a
Department of Biologic and Materials Sciences, School of known regulator of papilla development but details about
Dentistry, University of Michigan Ann Arbor, MI, USA locations of ligand, target responding cells and transcrip-
tional activators for Shh signaling are not known. We used
We have recently reported that both Wnt1-Cre and P0-Cre
immunostaining, in situ hybridization and reporters for Shh,
labeled neural crest derived cells are located in tongue epi-
Ptch1, Gli1 and Gli2- expressing cells to identify proliferating
thelium, taste papillae and taste buds, suggesting a neural
and differentiating cells in embryonic, postnatal and adult
crest contribution to taste bud cells. However, the difference
tongue, in papilla placodes, fungiform papillae and/or taste
is profound between Wnt1-Cre and P0-Cre lines in propor-
bud cells that participate in Shh signaling. Whereas there is a
tions and distribution patterns of neural crest derived cells
progressive restriction in location of the Shh ligand, a recep-
in tongue epithelium, i.e., abundant P0-Cre labeled epithelial
tive surround of Ptch1 and Gli1 expression in responding
cells versus few cells in Wnt1-Cre mouse. Such differences in
cells is maintained in particular epithelial and mesenchymal
distributions between Wnt1-Cre and P0-Cre labeled cells are
signaling centers throughout papilla development and taste
also found in other systems. To better interpret the data using
bud differentiation. From lineage tracing, we know that Gli1-
Wnt1-Cre and P0-Cre mouse lines for neural crest derivation
expressing cells and their progeny are located in fungiform
assays, we made a detailed expression analysis of the P0-Cre
papilla basal cells, in perigemmal cells and mesenchymal
transgenic mouse line at early embryonic stages in compari-
cells of the papilla core, and are progenitors of taste cells.
son with Wnt1-Cre using an R26R reporter. We found that
Further, using a doxycycline-regulated bitransgenic GLI2*
P0-Cre driven reporter expression emerges in the cranial neu-
mouse, in a functional test of activated Shh signaling in
ral crest region at E8.5 (6-somite stage) and is more profound
postnatal tongue epithelium, there is loss of filiform papilla
in hindbrain than midbrain. This is in contrast to the distribu-
spines and loss of fungiform papillae and taste buds. Loss of
tion pattern of Wnt1-Cre labeled neural crest cells, which is
papilla organs is accompanied by proliferation in suprabasal
extensive in midbrain but with far fewer cells in hindbrain.
layers of the lingual epithelium. The synthesized data posi-
The P0-Cre labeled cells are largely co-localized with neural
tion Shh signaling in multiple centers that are essential to
crest stem cell markers p75, Slug, AP2 and Sox 9. Combined
placode and papilla development, and to postnatal papilla
with our findings that P0-Cre labeled cells are far more abun-
and taste bud differentiation and maintenance. Shh roles are
dant than Wnt1-Cre in lingual epithelium, including taste
most likely via paracrine mechanisms, and engage epithelial/
papillae and taste buds, the data suggest that Wnt1 and P0
mesenchymal interactions. Acknowledgements: NIH Grants
mark overlapping but different subpopulations of neural crest
NIDCD DC000456 (CMM), NIDDK DK065850 (DLG),
cells, and that the hindbrain is the primary region of neural
NCI CA087837 (AAD).
crest cells that contribute to taste papillae and taste buds.
Acknowledgements: NIDCD NIH Grant R03DC009055 and
R01DC012308 to HXL, NIDCD R01DC000456 to CMM, #P223 POSTER SESSION V: HUMAN TASTE
NIDCR Grant R01DE020843 to YM PSYCHOPHYSICS; OLFACTION RECEPTORS;
TASTE DEVELOPMENT
#P222 POSTER SESSION V: HUMAN TASTE BDNF is Required for the Development of Adult Taste Bud
PSYCHOPHYSICS; OLFACTION RECEPTORS; Number and Normal Behavioral Responses to Sour Stimuli
TASTE DEVELOPMENT Abigail B. Menefee1, Robin F. Krimm2
Multiple Shh Signaling Centers in Embryo and Adult 1
dupont Manual High School Louisville, KY, USA, 2Dept.
Participate in Fungiform Papilla and Taste Bud Formation of Anatomical Sciences and Neruobiology, Univeristy of
and Maintenance Louisville Medical School Louisville, KY, USA
Hongxiang Liu1, Alex Ermilov2, Marina Grachtchouk2, Libo Brain derived neurotrophic factor (BDNF) regulates gusta-
Li1, Deborah L Gumucio3, Andrezej A Dlugosz2,3, Charlotte M tory system development. Because BDNF removal is neo-
Mistretta1 natal lethal, the long-term effects of BDNF removal on the
1
Department of Biologic and Materials Sciences, School structure and function of the adult gustatory system are
of Dentistry, University of Michigan Ann Arbor, MI, USA, unclear. To address this issue we examined the adult taste
system in conditional Bdnf knockouts in which Bdnf expres- preference tests to concentration series of NaCl, sucrose,
sion is reduced to one-tenth normal levels in the entire animal quinine, and citric acid. Terminal field volumes for each
(Bdnf lox/lox) and is completely removed from the lingual epi- nerve were significantly greater (2X – 4X) in early sodium-
thelium (K14-Cre;Bdnf lox/lox). K14-Cre;Bdnf lox/lox mice had restricted mice, and the overlapping zone that received all
very few fungiform taste buds remaining (11 ± 2) compared three nerves was over 15X greater in restricted mice. The dif-
to wild type (52 ± 5, p ≤ 0.002) or Bdnf lox/lox mice (42 ± 10; p ferences in terminal fields were accompanied by increased
≤ 0.02). The K14-Cre;Bdnf lox/lox circumvallate papillae con- preferences to NaCl and decreased aversions to citric acid,
tained 25% fewer taste buds than the control genotypes (p ≤ no differences in CT and IX whole-nerve taste responses,
0.025). There was no difference in taste bud number between and no differences in number of GSP, CT, and IX ganglion
wild type and Bdnf lox/lox, even though Bdnf lox/lox mice have cells. We conclude that the early dietary manipulation had
substantially reduced Bdnf expression. Therefore, as long as a profound effect on early NST development and that the
some BDNF remains, normal taste bud numbers are main- terminal field alterations impacted taste-related behaviors.
tained. Short-term lick rate tests of K14-Cre;Bdnf lox/ lox, Bdnf Acknowledgements: R01 DC00407
lox/lox
, and wild type mice were used to examine taste function.
Surprisingly, in spite of the large reduction in taste bud num-
ber, there was no statistical difference among the genotypes #P225 POSTER SESSION V: HUMAN TASTE
in lick rates to sucrose, quinine, and NaCl. This indicates PSYCHOPHYSICS; OLFACTION RECEPTORS;
that normal behavioral taste responses can be maintained in TASTE DEVELOPMENT
mice with few fungiform taste buds. However, K14-Cre;Bdnf
lox/lox
mice have higher lick rates to citric acid at pH=3.2 (p ≤ Renewal Kinetics of Taste Bud Cells in Adult Mice
0.024) and pH=2.8 (p ≤ 0.01) compared to wild type mice. Linda A. Barlow1,2, Brendan W. Ross1,2, Lauren A. Gross1,2
This indicates that removal of BDNF may cause a specific 1
Dept of Cell & Developmental Biology, University of
deficit in sour taste, which cannot be explained simply by the
Colorado School of Medicine Aurora, CO, USA, 2Rocky
loss of taste buds. Acknowledgements: DC007176
Mountain Taste & Smell Center, University of Colorado
School of Medicine Aurora, CO, USA
#P224 POSTER SESSION V: HUMAN TASTE Taste bud cells are continuously renewed from a population
PSYCHOPHYSICS; OLFACTION RECEPTORS; of progenitor cells, which express cytokeratin (K)5. These
TASTE DEVELOPMENT progenitors reside outside taste buds and give rise to daugh-
ter cells, which exit the cell cycle, enter buds and differenti-
Reorganization of Primary Afferent Terminal Fields in ate into one of 3 taste cell types (I, II, III). Differentiated
the Mouse Brainstem Produced by Early Prenatal Dietary taste cells live, on average, for 10 days, although variance
Sodium Restriction around this mean is large, suggesting different cell types have
Chengsan Sun, David L Hill different longevities (Beidler & Smallman ′65 J Cell Biol
27:263). To explore the idea of cell type-specific lifespans,
University of Virginia/Psychology Charlottesville, VA, USA
we employed inducible genetic birthdating, comprising a
Age-related decreases in terminal field volumes of the rat K5rtTA driver and a tetracycline- inducible reporter allele,
GSP, CT, and IX nerves and their overlapping fields in which encodes a nucleosomal protein, Histone2B fused with
the nucleus of the solitary tract (NST) occur during nor- GFP (tetO-H2BGFP, Tumbar et al., 2004 Science 303:359).
mal development. The processes involved in “pruning” the When K5rtTA;tetO-H2BGFP mice eat doxycycline (dox)
three terminal fields can be altered significantly when rats chow, K5+ cells produce H2BGFP, which is incorporated
are fed a sodium-restricted diet from E3-E12. All terminal into nucleosomes during S phase. Once mice are taken off
fields are relatively large during early postnatal ages and dox, H2BGFP is no longer transcribed, and the cohort of
thereafter fail to “prune”. Surprisingly, many of the termi- GFP labeled cells comprises the “pulse”. Here, bigenic mice
nal fields in restricted rats expand after 35 days postnatal. were fed dox chow for 12 hours, and tongues harvested
Thus, a very early period of dietary sodium restriction leads between 3 and 28 days. In the posterior circumvallate papilla,
to a late-onset expansion of terminal fields in the rat NST. an average of 2 cells/ bud was GFP+ after a 3 day chase.
To begin identifying the cellular/molecular mechanisms This increased to 4 GFP+ cells/bud by 14 days, and per-
responsible for this brainstem plasticity, we explored the ter- sisted through 21 days post- dox. At 28 days, however, only
minal field organization in adult mice that either received a 2 cells/bud were GFP+. At 14 days, 2 GFP+ cells/bud were
sodium- replete diet throughout development (controls) or PLCß2+ type II cells, whereas on average, less than 1 GFP+
mice fed the sodium-deficient diet from E3-E12. Moreover, cell/ bud were Snap25+ type III cells. The number of type
we counted the number of ganglion cells, representing the II cells/bud began to decline at 21 days, while type III cells/
three nerves, recorded whole-nerve neurophysiological taste bud were fewer by 28 days. We are currently determining if
responses from the CT and IX, and conducted 48hr. 2-bottle type II cell lifespan is less than that of type III cells, and/or
if type III cells are generated less frequently from later divi- contain a mixture of various taste active glycosides, with ste-
sions of GFP+ K5+ progenitors. Acknowledgements: R01 viol being the most abundant. The second most abundant
DC012383 to LAB P30 DC004657 to D. Restrepo glycoside is Rebaudioside A (RebA). RebA (>98% pure) is a
GRAS (generally recognized as safe) ingredient in the United
States, while stevia (the mixed extract) is classified as a dietary
#P226 POSTER SESSION V: HUMAN TASTE supplement. Another glycoside of interest is rebaudioside
PSYCHOPHYSICS; OLFACTION RECEPTORS; D (RebD), due to its high dose response for sweetness and
TASTE DEVELOPMENT minimal bitterness relative to steviol and RebA. Like saccha-
Bitter taste similarities among heterozygous MZ twins rin and acesulfameK (AceK), the bitterness from stevia gly-
compared with homozygous MZ twins. cosides varies across people. Previously, variable saccharin/
AceK bitterness has been associated with single nucleotide
Suzie Alarcon1, Ashley Sharples1, Paul A. S. Breslin1,2 polymorphisms (SNPs) in bitter receptor genes (TAS2Rs).
Rutgers, The State University of New Jersey New Brunswick,
1 As part of an ongoing study, we explored whether TAS2R
NJ, USA, 2Monell Chemical Senses Center Philadelphia, PA, USA SNPs may explain variable RebA and RebD bitterness. After
a brief orientation with bitter, sweet and metallic training ref-
Heterozygous alleles of select genes are known to be expressed erences, participants rated RebA, RebD, aspartame, sucrose,
differentially in different individuals. And for some genes, expres- and gentiobiose (a β 1–6 linked disaccharide) for these sensa-
sion in heterozygous monozygotic (MZ) twins is under strong tions on a general Labeled Magnitude Scale. Salivary DNA
genetic control (twins resemble each other in allele expression, was obtained and genotyped via Sequenom MassARRAY.
but differ from twin pair to twin pair). The mechanism of action As expected, mean RebD bitterness was much lower than
of differential allelic expression is unclear. In order to investi- RebA. In preliminary analyses, RebA bitterness associated
gate the potential genetic influence of differential expression of with a coding SNP in TAS2R9 and a synonymous SNP in
taste genes, we examined the bitter taste phenotypic response TAS2R50. For the TAS2R9 Ala187Val SNP, Ala187 allele
of MZ (identical) twin pairs to the bitter tastant 6n -propyl- carriers reported less bitterness than Val187 homozygotes.
2-thiouracil (PROP). Volunteers at the Twinsburg Twins Days For RebD, the TAS2R50 SNP predicted bitterness, although
Festival in Cleveland OH tasted PROP and recorded their this is likely a tag SNP for another polymorphism given the
perceived bitterness intensity on a generalized labeled magni- synonymous substitution. Finally, TAS2R31 SNPs previ-
tude scale (LMS). DNA samples were collected by cheek swab ously shown to predict AceK and saccharin bitterness did not
from each subject and were genotyped at TAS2R38 amino acid predict bitterness from RebA or RebD. Acknowledgements:
codon positions 49, 262, and 296. MZ twins were grouped by Supported by funds from the Pennsylvania State University
their TAS2R38 diplotype. We compared the similarity of Twin and NIH grant DC0010904.
A vs Twin B for heterozygous MZ twins to the similarity of
Twin A vs Twin B for homozygous MZ twins. The heterozy-
gous MZ group displayed greater perceptual variation between
twins than the homozygous MZ group, despite the fact that the #P228 POSTER SESSION V: HUMAN TASTE
homozygous group was comprised of both AVI/AVI and PAV/ PSYCHOPHYSICS; OLFACTION RECEPTORS;
PAV homozygous subgroups. Thus, to the degree that pheno- TASTE DEVELOPMENT
type reflects expression, it appears that allelic expression of Sucrose analgesia and the cold pressor test in young men:
bitter taste receptors is under significant non-genetic control Methodological considerations
within heterozygous MZ twins. Acknowledgements: Funded in
Rachel G Antenucci1,2, John Prescott3, Theresa L White4, John
part by NIH DC 02995 to PASB.
E Hayes1,2
1
Department of Food Science, The Pennsylvania State
#P227 POSTER SESSION V: HUMAN TASTE University University Park, PA, USA, 2Sensory Evaluation
PSYCHOPHYSICS; OLFACTION RECEPTORS; Center, The Pennsylvania State University University Park,
TASTE DEVELOPMENT PA, USA, 3TasteMatters Research & Consulting Sydney,
TAS2R polymorphisms and the bitterness of RebaudiosideA Australia, 4Department of Psychology, LeMoyne College
and RebaudiosideD Syracuse, NY, USA
Alissa L. Allen1, John E. McGeary 2, John E. Hayes1 Sucrose is mildly analgesic in infant rats, human neo-
nates, and prepubertal children. This effect generalizes
1
Department of Food Science, Penn State University Park,
to non-nutritive sweeteners, indicating the effect is due to
PA, USA, 2Providence VA Medical Center Providence, RI, USA
sweet taste. However, consistent sweet analgesia in adults
With growing demand for natural non-nutritive sweeten- remains elusive: some studies find an effect while others do
ers, extracts from the Stevia rebaudiana Bertoni plant have not. Pain can be safely induced in the laboratory using the
become a popular replacement for sugar. These exacts Cold Pressor Test (CPT), providing a convenient means
to study this phenomenon. It is unresolved whether the mouth ratings for sucrose, quinine, and PROP. We char-
inability to consistently observe analgesic effects in adults acterized subjects as high affect using both approaches.
is due to methodological issues associated with the CPT Regardless of the categorization method, sucrose, quinine,
or an age dependent effect. White and Prescott (AChemS, and PROP intensity ratings did not differ by group in
2012) reported strong order effects; tolerance was greater ANOVA. When groups were predicted in logistic regres-
in the second CPT within a session. They suggested sion (‘do higher taste ratings predict being a foodie?’)
rewarming the hand between tests might reduce this bias. there was no evidence supporting this hypothesis. Adding
Here, we describe 2 studies that attempt to refine an adult the personality trait Sensation Seeking as a covariate did
sweet analgesia CPT paradigm. In study 1 (36 men), pain not alter these conclusions. We did find evidence of lower
was induced by placing the dominant hand in circulating Sensation Seeking scores among foodies (as defined via the
water at 8oC. Within a session, tastants were water and difference score), but further inspection suggests this was
0.7M sucrose; participants were tested twice (fed / fasted) due to a small increase in the mean liking of pleasant non-
in a double crossover design. The hand was rewarmed in food items when mean food liking was flat. These data fail
35oC water between trials, and hand temperature was con- to support the hypotheses that a) hypergeusic individuals
firmed with a laser thermometer. No tastant effect, regard- show higher food related affect or that b) higher food affect
less of hunger state, was observed. However, a weak trend predicts heightened taste response. Acknowledgements:
for an order effect within a session was evident, despite funds from the Pennsylvania State University and NIH
rewarming. In study 2 (27 men), subjects received sucrose grant DC0010904.
or water on separate days after an overnight fast. The cold
bath was reduced to 4oC, and outcomes included pain
threshold (onset in s) and tolerance (hand withdrawl in #P230 POSTER SESSION V: HUMAN TASTE
s). Pain thresholds and tolerance were greater on day 2, PSYCHOPHYSICS; OLFACTION RECEPTORS;
but this occurred irrespective of tastant. Collectively, these TASTE DEVELOPMENT
data suggest increased tolerance in the CPT with repeat
exposure is not an effect of initial hand temperature. Effects of Food Neophobia on Salivary ph, Cortisol and
Acknowledgements: The Pennsylvania State University Adrenal Level
and USDA Hatch Act funds August Capiola, Bryan Raudenbush, Amanda Schultz
Wheeling Jesuit University Wheeling, WV, USA
#P229 POSTER SESSION V: HUMAN TASTE Food neophobics (individuals reluctant to try novel foods)
PSYCHOPHYSICS; OLFACTION RECEPTORS; and food neophilics (individuals with an overt willingness
TASTE DEVELOPMENT to try novel foods) differ in several physiological aspects.
Phenylthiocarbamide (PTC) tasting ability, a genetic pre-
Are individuals with elevated food liking scores (‘foodies’) disposition, differs among the two groups with more food
hypergeusic? neophobics possessing this inherited trait. Food neophobics
Nadia K Byrnes, Alissa L Allen, Emma L Feeney, Rachel J salivate less when presented with novel foods and have higher
Primrose, John E Hayes physiological stress responses to novel foods (increased pulse,
GSR, and respirations). The present study assessed salivary
Department of Food Science University Park, PA, USA
pH, adrenal level and cortisol level in food neophobics, food
The public believes that foodies are supertasters (and vice neophilics and an average group, to determine whether such
versa), consistent with reports that chefs and wine experts salivary flow and physiological stress reactions could par-
report greater bitterness from propylthiouracil (PROP). tially be explained by such variables. Salivary mouth swab
However, the two reports that test this hypothesis con- samples were obtained from 117 participants, who also com-
flict. Minski et al. reported ‘high food interest’ individu- pleted the Food Neophobia Scale (FNS) to assess level of
als (‘foodies’) rated quinine, sodium chloride and PROP as food neophobia. A significant MANCOVA result was found,
more intense than those with average or low food interest F=2.47, p=.03. Further analysis revealed food neophobics
in a laboratory study (n=100), while Pickering et al. failed had significantly higher levels of salivary cortisol compared
to find any difference in the bitterness of PROP impreg- to food neophilics and the average group, F(2,102)=7.53,
nated discs sent via mail (n>900). These studies also differ p=.001. The finding that higher levels of the stress hormone
in how high food affect individuals were identified: via a cortisol are present in food neophobic’s saliva supports past
ratio of affective ratings for all foods to pleasant non-food research indicating a greater physiological stress reaction
item versus a difference score of mean liking for all foods to novel food stimuli in these individuals. Future research
minus mean liking for all non-foods. Here, we explore should assess whether exposure to novel foods can decrease
this question in 246 subjects who completed a general- the level of salivary cortisol in food neophobics, as a way of
ized hedonic survey (i.e. food & nonfood items) and whole promoting a more varied and healthful diet.
#P231 POSTER SESSION V: HUMAN TASTE Institutes of Health, under contract number HHS-N-260-
PSYCHOPHYSICS; OLFACTION RECEPTORS; 2006-00007-C. The National Children’s Study also provided
TASTE DEVELOPMENT support for norming.
The NIH Toolbox Brief Gustation Assessment Protocol

Susan E. Coldwell1, Valerie B. Duffy2, Linda Bartoshuk3, #P232 POSTER SESSION V: HUMAN TASTE
James W. Griffith4, Howard J. Hoffman5 PSYCHOPHYSICS; OLFACTION RECEPTORS;
TASTE DEVELOPMENT
1
University of Washington School of Dentistry Seattle,
WA, USA, 2University of Connecticut College of Establishing Best Practices for an Effective Community-
Agriculture and Natural Resources Storrs, CT, USA, Based Research Laboratory
3
University of Florida College of Dentistry Gainesville,
FL, USA, 4Northwestern University Feinberg School of Tiffany M. Derr1, Patty McNamara2, Diana Boyles1, Brian
Medicine Evanston, IL, USA, 5National Institutes of Health, Hostetler1, Nicole L. Garneau1
National Institute on Deafness and Other Communication 1
Denver Museum of Nature & Science Denver, CO, USA,
Disorders Bethesda, MD, USA 2
Independent Consultant Chicago, IL, USA
NIH Toolbox developed standardized, brief assessments
for sensory, motor, cognitive and emotional function that The American Association for the Advancement of Science
are designed for use in longitudinal studies, epidemiologi- has become a strong advocate for positive and effective
cal research, and clinical trials. The sensory battery consists dialogue between the scientific and public communities,
of brief assessments of gustation, olfaction, vision, audi- but recognizes the strengths, weaknesses and limitations
tion, pain, and vestibular balance; all six can be admin- that exist towards these efforts. As relayed in the AAAS
istered within 30 minutes, including 6 minutes for the Public Engagement online video, “Researchers may not
assessment of gustation. The Gustation Assessment begins be accustomed to such public interaction, but it’s crucial
with instructions in use of the general Labeled Magnitude to the science/society relationship.” Towards this effort,
Scale by rating of remembered lights (dimly lit restaurant, the Denver Museum of Nature & Science established a
well-lit room, brightest light ever seen). Four taste trials are citizen-scientist run, community-based Genetics of Taste
then delivered: 1 mM Quinine HCl applied to the anterior laboratory. In the past three years of active research, we’ve
tongue, 1 M NaCl applied to the anterior tongue, 1 mM found that the community-based lab is a unique environ-
Quinine HCl whole mouth (sip and spit), and 1 M NaCl ment for conducting research in which we are constantly
whole mouth. Rinsing with water is done between trials. optimizing the balance between providing an enjoyable
As part of the NIH Toolbox national norming study, the educational experience and collecting data that is scien-
Gustation Assessment was given to 1843 English-speakers tifically sound. Here we present the lessons learned in
and 240 Spanish- speakers. These included 494 subjects developing a community-based lab and how academic and
aged 12 to 15 years and 509 aged 15 to 19 years. For 172 research institutions can apply this model to their studies.
subjects, the battery was given twice to establish test-retest In terms of research study design, involvement of citizen-
reliability. Preliminary intraclass correlations (ICC) indicate scientists in the planning process is not only a suggested
that the test is reliable for whole mouth ratings (ICC = 0.54 best practice, but also helps to establish motivation and
for Quinine and 0.57 for NaCl) and reliable for NaCl on commitment early in the study. Initial planning should
the anterior tongue (ICC = 0.42). Quinine ratings for the include in-depth discussions of intended outcomes and a
anterior tongue were less reliable (ICC = 0.29), but this was clearly defined timeline with tangible milestones to meas-
always the first trial. For the entire norming sample, small, ure progress. Educational outcomes should be included,
but statistically significant, declines in taste intensity with and care should be taken to have these goals as clearly
age were observed for each of the four trials (Pearson corre- defined as the research outcomes. Finally, an investment
lations from −0.1 to −0.2, p <0.001). Initial findings indicate to rigorously train citizen- scientists is a large upfront
that this brief gustation assessment is reliable and sensitive time commitment for professional staff, but we have found
to the gradual decline in taste perception that occurs with that this investment ensures long term quality control of
age. Acknowledgements: This project was funded by Federal citizen-science collected and prepared data, which saves
funds from the Blueprint for Neuroscience Research and the resources in the long term. Acknowledgements: 2008–
Office of Behavioral and Social Sciences Research, National 2012 R25 RR025066 NIH NCRR SEPA
#P233 POSTER SESSION V: HUMAN TASTE between sweet taste and obesity, and the overlap of sweet and
PSYCHOPHYSICS; OLFACTION RECEPTORS; umami tastes at mechanistic level (e.g., both employ the taste
TASTE DEVELOPMENT receptor T1R3 in the dimer), the present study, examined the
perception of these two taste qualities along with salt (as a
Carbonic anhydrase CA6 (gustin) polymorphisms and control taste) in obese and normal-weight children. To this
perceived taste intensity end, 97 children were phenotyped for detection thresholds for
Emma L Feeney1, John E McGeary2, John E Hayes1 sweet (sucrose), savory (monosodium glutamate), and salty
(NaCl) taste using age-appropriate psychophysical testings,
1
Department of Food Science, Pennsylvania State University
dietary habits, blood pressure and obesity. Preliminary analy-
State College, PA, USA, 2Providence VA Medical Center
ses revealed that the detection threshold for sucrose, salt, and
Providence, RI, USA
MSG are similar to that previously reported in adults and
Polymorphisms in the carbonic anhydrase CA6 gene (i.e., there were no differences observed between normal weight
gustin) reportedly account for additional variation in the bit- children and obese children in the detection thresholds for
terness of 6-n-propylthiouracil (PROP) above and beyond any of these basic tastes. In normal- weight, but not obese
that of the TAS2R38 gene. It has been hypothesized that children, salt detection thresholds were positively correlated
CA6 influences taste thresholds for and the perceived inten- with systolic blood pressure. As a group, the greater the waist
sity of PROP via differences in gustin function, and that gus- circumference, the lower the sucrose detection threshold (the
tin may be a trophic factor for taste bud development (Calò more sensitive the child was to sucrose) (p=0.02). No such
et al 2011). If true, we reasoned such effects should not be relationships existed for salt or MSG in children. Whether
limited to PROP, and should generalize across qualities to the lower detection thresholds for sucrose are associated
other stimuli. As part of an ongoing study, healthy partici- with stronger reinforcing value of sweet foods, as has been
pants (aged 18–45) rated their perception of sweet, sour, salty, observed in adults, and understanding the link between salt
savory/umami, bitter, and burning sensations on a general- taste thresholds and blood pressure, are important areas for
ized Labeled Magnitude Scale for sucrose, sodium chloride, future research. Acknowledgements: This project was funded
potassium chloride, quinine hydrochloride, PROP, citric acid, by an investigator-initiated grant from Ajinomoto, Inc.
capsaicin and a monosodium glutamate/inosine monophos-
phate (MSG/IMP) mixture. Fungiform papillae density
was assessed via digital still microscopy. Salivary DNA was #P235 POSTER SESSION V: HUMAN TASTE
obtained and genotyped using Sequenom MassARRAY. We PSYCHOPHYSICS; OLFACTION RECEPTORS;
examined 14 CA6 SNPs: rs12748400, rs17032907, rs2274327, TASTE DEVELOPMENT
rs2274328, rs2274333, rs2274334, rs3737665, rs3765964,
rs3765965, rs3765967, rs3765968 and rs7545200. We failed to Statistical Analysis of Factors Previously Described as
observe differences in sweet, sour, or savory ratings across any Significant in the Ability to Taste Propylthiouracil Yields
of the SNPs tested. Nor were there any differences in FP den- Roles for Age, Sex and Tas2r38 Haplotype, but not
sity for any of the SNPs examined. However, perceived salti- Fungiform Papillae Density
ness associated with a number of SNPs in CA6. Additionally, Nicole L. Garneau, Tiffany Derr
FP density was correlated with the intensity of sucrose, PROP
Denver Museum of Nature & Science Denver, CO, USA
and salt, as would be expected. In summary, polymorphisms
within CA6 did not predict overall differences in taste inten- The Genetics of Taste research study at the Denver Museum
sity, although salty intensity differed with some CA6 SNPs. of Nature & Science is in a unique position to collect samples
We did not find evidence to support an association between from a diverse population across a wide age range. Using this
CA6 SNPs and PROP bitterness that had been reported pre- large population sample we set out to establish the scientific
viously. Acknowledgements: Supported by funds from the credibility of our community-based laboratory by replicat-
Pennsylvania State University and NIH grant DC010904. ing four previously reported statistically significant factors in
the ability to taste propylthiouracil; 1)Age, 2)Sex, 3)Tas2r38
#P234 POSTER SESSION V: HUMAN TASTE haplotype, and 4)Fungiform papillae density. Using regres-
sion analysis and the Student T-test we can replicate the role
of age and sex in taste score (gLMS following a propylthi-
TASTE DEVELOPMENT
ouracil taste test). Similarly, the presence of at least one
Taste Detection Threshold in Children dominant allele for the Tas2r38 gene is a significant predictor
Susana Finkbeiner, Julie A. Mennella, Loma B. Inamdar of taste. Finally, in order to decrease subjectivity, we devel-
oped a dichotomous key for objective analysis of fungiform
Monell Chemical Senses Center Philadelphia, PA, USA papillae density. Using this method we did not find that
In light of the unknown relationship between taste per- increased papillae density correlates to an increased pro-
ception and obesity in children, the uncertain relationship pylthiouracil taste score. In conclusion, the ability to replicate
the significance of age, sex and Tas2r38 haplotype, as well on the T1R2-T1R3 receptor and raise new questions about the
as the development of objective methodology for papillae factors that can affect excitatory and inhibitory interactions
analysis, all demonstrate the ability for citizen-scientists in a between these sites. Acknowledgements: Supported in part by
community- based laboratory to collect, prepare and analyze NIH grant DC005002
data that can contribute to the field of chemoreception. In
addition, we submit that using standardized methodology
for fungiform papillae density allows for a more objective
analysis of morphological data. Using this methodology we PSYCHOPHYSICS; OLFACTION RECEPTORS;
find no relationship between fungiform papillae density and TASTE DEVELOPMENT
propylthiouracil score in our data set. This data contradicts Language Determines Whether Taste Sensitivity is Related
previously published studies and suggests that fungiform to Moral Disgust
papillae density may not be as reliable a metric for classify-
Rachel S, Herz
ing taster status as previously thought. Acknowledgements:
This work was supported by volunteer citizen-scientists at the Department of Psychiatry and Human Behavior Providence,
Denver Museum of Nature & Science, and through support RI, USA
from 2008–2012 from R25 RR025066 NIH NCRR SEPA. The stimuli that trigger emotional disgust are currently
debated. Most contested is whether responses to visceral
triggers of disgust (e.g., cockroaches, disease) are fundamen-
#P236 POSTER SESSION V: HUMAN TASTE tally the same as responses elicited by moral transgressions
PSYCHOPHYSICS; OLFACTION RECEPTORS; (e.g., lying, cheating). To address this issue, the present study
TASTE DEVELOPMENT examined whether: (1) visceral and moral disgust share a
The effects of temperature on sequential and mixture common oral origin (the rejection of bitter tasting poisons);
interactions between sucrose and saccharin (2) verbal priming can alter whether disgust is experienced
as a function of taste sensitivity. 102 undergraduates com-
Barry G Green1,2, Danielle Nachtigal1 pleted a “Behavioral Situations Questionnaire” developed
The John B. Pierce Laboratory New Haven, CT, USA, 2Yale
1 for the present research which compared “grossed out” and
University School of Medicine New Haven, CT, USA “angry” to assess three types of moral transgressions that var-
ied in the degree to which a visceral disgust dimension was
The sweet taste of sucrose and saccharin has been shown to
invoked (non-visceral, implied visceral, directly visceral), and
depend on stimulation of the T1R2-T1R3 receptor, but it is
two standard tests of disgust sensitivity. After completing
also clear that these two stimuli interact with the receptor in
the questionnaires, taste sensitivity was assessed with a bit-
different ways. Most recently it was found that self-adaptation
ter tasting compound (6-n-propylthiouracil; PROP). Results
is temperature-dependent for sucrose but not for saccharin
showed that the more bitter PROP tasted the more sensitive a
(Green & Nachtigal, 2012), and a previous study showed that
participant was to visceral measures of disgust sensitivity, but
high concentrations of saccharin can block the sweetness of
not to moral disgust sensitivity. There were also no effects for
sucrose (and of itself) and evoke a sweet water taste (Galindo-
taste sensitivity or type of moral transgression when “angry”
Cusperino et al, 2006). The aim of the present study was to
was primed for evaluating the transgressions. However, when
determine if temperature modulates the ability of sucrose to
“grossed out” was primed, the more intense PROP tasted the
cross-adapt saccharin and/or of saccharin to block the sweet-
more “grossed out” participants were by all transgressions,
ness of sucrose and produce a sweet water taste. Subjects rated
regardless of their visceral nature. This supports the proposi-
the sweetness and bitterness of 0.42 M sucrose, 3.2 mM saccha-
tion that moral and visceral disgust do not share a common
rin, 100 mM saccharin, or binary mixtures of sucrose and the 2
oral origin, but shows that linguistic priming can transform a
concentrations of saccharin, with and without pre-exposure to
moral response into a viscerally repulsive event and that sus-
themselves or each of the other stimuli. The variables of inter-
ceptibility to this priming varies as a function of an individu-
est were the duration of pre-exposure (3 or 10 s) and solution
al’s sensitivity to the origins of visceral disgust— bitter taste.
temperature (37° or 21°C). The stimuli were sampled by dip-
ping the tongue tip into the solutions, and intensity ratings were
made on the gLMS before the tongue was retracted back into #P238 POSTER SESSION V: HUMAN TASTE
the mouth. The results confirmed the previous findings and PSYCHOPHYSICS; OLFACTION RECEPTORS;
showed that (1) the magnitude of sweet water taste (after expo- TASTE DEVELOPMENT
sure to 100 mM saccharin) is temperature-dependent, and (2)
Individual differences in the rate of salivary α-amylase
surprisingly, pre- exposure to sucrose for 3 or 10 sec appeared to
production and its role in the perception of glucose
counteract the ability of 100 mM saccharin to block sweetness,
polymers
independent of temperature. These results support the hypoth-
esis that sucrose and saccharin bind to at least 2 different sites Trina J. Lapis, Michael H. Penner, Juyun Lim
Oregon State University Corvallis, OR, USA the sweet taste of sugars. The current study was therefore
Our recent data suggest that some humans can consistently designed to measure individual differences in taste percep-
taste glucose polymers, implying possible existence of a glu- tion of various carbohydrates (glucose, sucrose, and glucose
cose polymer receptor. A potential confound of this idea is polymers of different chain length) and NaCl. Ss rated taste
that the hydrolysis byproducts of the glucose polymers, i.e., intensity of test solutions while their noses were clamped.
glucose and/or maltose, may have, at least to some extent, The results showed that the perceived intensities of glucose,
influenced taste responsiveness of the tasters. The latter sucrose, and NaCl were significantly correlated (r=.68~.80,
scenario is easily rationalized due to the catalytic activity p<.01), but not with the glucose polymers, whereas the inten-
of salivary a-amylase. The present study was designed to sity ratings of the glucose polymers were highly correlated
investigate (1) individual differences in the rate of α-amylase with one another (r=.62~.86, p<.01). Notably, there was
production and (2) the role that it may play in glucose poly- much greater variability across individuals in the responsive-
mer perception. Measured rate of salivary flow (mg/sec) ness to the glucose polymers than to the other stimuli. These
and α-amylase activity per mg saliva were used to calculate data provide evidence that some humans can consistently
the rate of α-amylase production (activity/sec). The same taste glucose polymers of various chain lengths (DE 20–5)
Ss rated the taste intensity of glucose, sucrose, and glucose and that the reactivity to glucose polymers is independent of
polymer solutions (differing in average chain length) follow- that to sugars. The current findings thus support the exist-
ing a sip and spit procedure. Results showed large individual ence of a secondary carbohydrate receptor in humans that is
differences in the rate of α-amylase production (>30-fold). activated by glucose polymers.
This can be attributed to the large differences in salivary flow
rate (>18-fold) and α-amylase activity (>30-fold). Notably,
average rates of α-amylase production were similar between #P240 POSTER SESSION V: HUMAN TASTE
the taster and non-taster groups. Further, within each group, PSYCHOPHYSICS; OLFACTION RECEPTORS;
responsiveness to glucose polymers did not appear to differ TASTE DEVELOPMENT
between individuals with high and low rates of α-amylase The role of gene expression in the human TAS2R38
production. These findings suggest that salivary α-amylase genotype-phenotype relationship
plays an insignificant role in glucose polymer perception.
Alternatively, it is possible that the chain lengths of the Sarah V. Lipchock1, Andrew I. Spielman2, Julie A. Mennella1,
glucose polymers tested were too short, i.e., the effect of Danielle R. Reed1
α-amylase on the hydrolysis was comparable between high Monell Chemical Senses Center Philadelphia, PA, USA,
1
and low α-amylase producers. This possibility is currently College of Dentistry, New York University New York, NY,
2
being explored in a follow-up experiment by using more USA

complex carbohydrates as test stimuli.
Bitter taste is important for sensing and avoiding toxins,
but the aversion to bitter can lead to low consumption of
#P239 POSTER SESSION V: HUMAN TASTE vegetables, an important source of phytochemicals. Single-
PSYCHOPHYSICS; OLFACTION RECEPTORS; nucleotide polymorphisms in the genes that code for the 25
TASTE DEVELOPMENT human bitter receptors (TAS2Rs) result in different indi-
vidual responses to taste stimuli. One example of this phe-
Evidence that humans can taste glucose polymers nomenon is the TAS2R38 gene, which encodes a receptor
Juyun Lim, Trina J. Lapis that recognizes compounds including 6-n-propylthiouracil
(PROP) and goitrin, found in broccoli. Cell-based assays
Oregon State University Corvallis, OR, USA and psychophysical threshold measurements confirm that
Previous findings of behavioral and electrophysiologi- receptor with the amino acid sequence PAV responds to
cal studies have shown that rodents can taste solutions of PROP, while the AVI form does not. Heterozygous (PAV/
Polycose and further can discriminate the taste of Polycose AVI) individuals display a broad range of abilities to taste
from that of sugars. Recent studies have also shown that PROP. To examine the role of allele-specific gene expres-
the T1R2/T1R3 single and double KO mice respond signifi- sion in the TAS2R38 genotype-phenotype relationship we
cantly to Polycose. Based on these data, the possible exist- recruited healthy, non-smoking adults with the PAV/AVI
ence of a glucose polymer receptor has been proposed. In diplotype (N=18). Psychophysical ratings for several bitter
contrast, it has been assumed that glucose polymers are compounds and vegetable juices were related to levels of
tasteless to humans, although it is known that they evoke TAS2R gene expression from human fungiform taste papil-
a slight odor. During a preliminary study of the role of lae. Increased levels of the PAV form of TAS2R38 were posi-
salivary α-amylase in the perception of glucose polymers, tively correlated with the subject’s intensity ratings for PROP
some Ss reported that the glucose polymers had “bread-” and broccoli juice. Expression of TAS2R43 was associated
or “cereal-like” taste which they could differentiate from with ratings for caffeine, urea, denatonium benzoate and
quinine. This relationship resulted from copy number varia- #P242 POSTER SESSION V: HUMAN TASTE
tion of TAS2R43 and suggests that TAS2R43 acts as a proxy PSYCHOPHYSICS; OLFACTION RECEPTORS;
for bitterness sensitivity from the cluster of receptors on TASTE DEVELOPMENT
chromosome 12. Our data provide a link between genotype
and phenotype in the taste system. Our work lays a foun- The Chemosensory Component in the 2012 National Health
dation for future studies about regulatory mechanisms and and Nutrition Examination Survey (NHANES): Test-Retest
implications of taste receptor expression as it relates to diet Analysis and Comparison with Laboratory-Based Measures
and vegetable consumption in humans. Acknowledgements: Shristi Rawal1, Howard J Hoffman2, Kathy Bainbridge2,
Supported by F32DC011975 (SVL), P30DC011735 (DRR) Valerie B Duffy1
and R01DC011287 (JAM). 1
University of Connecticut/Allied Health Sciences Storrs,
CT, USA, 2National Institute on Deafness and Other
Communication Disorders/ Division of Scientific Programs
#P241 POSTER SESSION V: HUMAN TASTE Bethesda, MD, USA
TASTE DEVELOPMENT The NHANES now includes a standardized chemosensory
protocol with brief assessments of taste intensity and odor
Evidence that cooling affects the sweetness of sugars via 2 identification conducted in a mobile exam center. After ori-
separate mechanisms. entation to the general Labeled Magnitude Scale and scaling
Danielle J. Nachtigal1, Barry G. Green1,2 intensity of LED-generated lights, participants report inten-
sities of 1 M NaCl and 1 mM quinine hydrochloride applied
The John B. Pierce Laboratory New Haven, CT, USA,
1
to the tongue tip and these plus 0.32 M NaCl with the whole
Department of Surgery, Yale University School of Medicine
2
mouth. Olfactory dysfunction is assessed with two 4-item,
New Haven, CT, USA
scratch- and-sniff tests (Pocket Tests™ (PT), Sensonics, Inc)
Previous experiments in our laboratory demonstrated that with food (strawberry, chocolate, onion, grape) and common
temperature can affect the sweetness of sucrose, glucose, (leather, soap, smoke, natural gas) odors. Here we examined
and fructose solutions by modulating taste adaptation: test-retest reliabilities of the chemosensory protocol, com-
Cooling from 37°C to 21°C significantly increased the rate pared the PT with an olfactometer identification task, and
and amount of self- adaptation when the tongue tip was added scaling to all odor tests. Fifty adults (mean age=27,
dipped into the solutions for several seconds. However, range 18–62 yrs), primarily Caucasian and Asian, college-
cooling to 21°C did not reduce initial sweet taste intensity, educated and reportedly healthy, were tested in a laboratory
and thus did not directly affect sensitivity to the sugars. twice within two weeks. The taste test intraclass correlations
In the present study we investigated whether cooling to (ICC, single measures) ranged from moderate to substantial
colder temperatures would further accelerate sweet taste agreement (0.47 NaCl, 0.50 quinine tongue tip; 0.75 quinine
adaptation and/or begin to reduce sweet taste sensitivity. whole mouth). Both PT trials classified 98% of the partici-
Subjects rated the sweetness of 0.42 M sucrose solutions at pants as normosmic. There were good correlations between
temperatures of 41°, 37°, 30°, 21°, 10° or 5°C after dip- the individual PT odor intensities (ranging 0.44 for grape to
ping the tongue tip for 0, 3, or 10 sec into either 0.42 M 0.72 for gas) and a moderate overall ICC (0.56). The PT and
sucrose or pure dH2O of the same temperature. Sweetness olfactometer odor intensities were correlated, averaging 0.5
ratings were made prior to retracting the tongue back into (ranging 0.28 for chocolate to 0.65 for grape). Correct and
the mouth. There were two main findings: (1) sweet taste incorrect odor identification was consistent across the PT and
adaptation increased significantly at 21°C (replicating our olfactometer tests, averaging 86% agreement. These findings
previous results), and (2) pre-exposure to pure dH O at 5° show that, in ideal testing situations, the NHANES protocol
or 10°C significantly reduced sweet taste intensity inde- had very good reliability and the Pocket Test corresponded
pendent of taste adaptation. These findings indicate that reasonably well among normosmics with a measure having
cooling interferes with the perception of sucrose sweet- more stimulus control. Acknowledgements: NIDCD/NIH
ness in two different ways: mild cooling increases the rate
of adaptation whereas more extreme cooling reduces the
initial sensitivity to sucrose. Experiments are planned that #P243 POSTER SESSION V: HUMAN TASTE
will test the hypothesis that these two effects occur at dif- PSYCHOPHYSICS; OLFACTION RECEPTORS;
ferent stages of the transduction cascade, with the former TASTE DEVELOPMENT
affecting the binding of sucrose to the transmembrane
Efficacy of sodium and glutamate in reducing bitterness in
region of the T1R2-T1R3 receptor, and the latter modu-
children and adults
lating a temperature-sensitive intracellular mechanism that
is common to G-protein coupled receptors (e.g., TRPM5). Kristi M. Roberts, Phoebe S. Mathew, Corrine J. Mansfield,
Acknowledgements: NIH grant RO1 DC005002 Danielle R. Reed, Julie A. Mennella
Monell Chemical Senses Center Philadelphia, PA, USA monosodium glutamate (MSG). However, sensitivity for the
A central challenge of administering medicine to children is disaccharide sucrose (furanose), and perhaps for the disac-
a ‘matter of taste’ because drugs, by their very nature, often charide maltose (pyranose), remains unchanged (Gonzalez,
taste unpleasant with bitter taste being the primary culprit. 2009; Kennedy et al., 2012). Here we conducted further test-
As part of an ongoing study on individual differences in bit- ing with maltose. Subjects rinsed their tongues with 4 mM
ter taste perception, we present here preliminary data on rat- Na-c or water for 10 sec once a day for 10 days. On day 11 or
ings of the bitterness of urea and propylthiouracil (PROP) 12, they tasted a concentration series of maltose, each con-
with and without the addition of a sodium gluconate or glu- centration paired with water, and indicated which of each
tamate (putative blockers) in a racially diverse group of 3- to pair was “the sweetener.” Subjects treated with Na-c showed
10-year old children and adults. Using forced-choice proce- increased sensitivity for maltose (p=0.004). With respect to
dures, each child and adult was presented with all possible the high-intensity sweeteners, responses to Na-c and sucra-
pairs of the four solutions for each bitter agent (e.g., 0.5M lose are increased by Na-c treatment while responses to
urea, 0.3M sodium gluconate, 0.5 M urea+0.3M sodium glu- D-tryptophan are decreased. Because responses to MSG
conate, and water), one pair at a time, and asked to indicate also are decreased after Na-c treatment, the effect may be
which of the pair tasted more bitter. The data for each bit- different for amino acid stimuli. Testing with the amino acid
ter stimulus were expressed as the proportion of children or sweetener aspartame is in progress. Human psychophysical
mothers that chose one member of the pair as tasting more and animal neurophysiology data suggest peripheral mecha-
bitter and from this, each of the four solutions were ranked nisms (Faurion et al., 2002; Hassan et al., 2006; Gonzalez
according to subject’s ratings (1=least bitter; 4=most bit- et al, 2009). We have suggested that binding of the treatment
ter). Adults also used gLMS to rate taste qualities of each compound with the receptor subunit T1R3 leads to changes
solution in another session. Preliminary analysis revealed in binding or other steps in the receptor response to the test
that most children (75%) were able to complete the task; the compound. Our results show that the mechanism(s) for the
vast majority of those who did not complete the task were changes affect(s) stimulation by various sweeteners differ-
of the younger age (<6 years) range, highlighting the dif- ently. The generalization of effects supports our hypoth-
ficulty in such assessments of younger children. In adults, esis that sweet compound interaction with T1R3 leads to
we also found that either blocker significantly reduced the changes in the overall receptor response. Acknowledgements:
gLMS rating of urea bitterness but sodium gluconate was Supported by NIH NIDCD R15DC009042 to LMK.
ineffective for blocking the bitterness of PROP. Paired com-
parison data yielded similar findings in adults as well as
children. Acknowledgements: This project was funded by #P245 POSTER SESSION V: HUMAN TASTE
R01DC011287 and supported by P30DC011735 from the PSYCHOPHYSICS; OLFACTION RECEPTORS;
National Institute on Deafness and Other Communication TASTE DEVELOPMENT
Disorders. The content is solely the responsibility of the Glutamate Detection Thresholds Are Altered by the
authors and does not necessarily represent the official views Addition of 5’-Ribonucleotides
of the NIDCD or the National Institutes of Health.
Ashley A Sharples1, Suzie Alarcon1, Paul AS Breslin1,2
1
Rutgers University New Brunswick, NJ, USA, 2Monell
#P244 POSTER SESSION V: HUMAN TASTE Chemical Senses Center Philadelphia, PA, USA
TASTE DEVELOPMENT Umami taste intensity of glutamate is synergisti-
cally increased in humans by the addition of inosine
Generalization of the effects of Na-cyclamate treatment 5′-monophosphate (IMP) disodium salt and guanosine
on sensitivity for sugars, sweeteners, and monosodium 5′-monophosphate (GMP) disodium salt, two puriner-
glutamate supports a role for T1R3 in experience-induced gic ribonucleotides. We hypothesized that the addition of
changes in taste these and other ribonucleotides would decrease the abso-
Julia Sabin1,2, Samer Halabiya2, Todd P Livdahl1, Linda M lute detection threshold of L-glutamic acid potassium salt
Kennedy1,2 (MPG) when in admixture; thus, sensitivity to glutamate
would be increased. We first measured the absolute thresh-
Clark University Worcester, MA, USA, 2University of
1
old to MPG and compared this to the threshold for MPG
Washington Seattle, WA, USA in the presence of a constant background level of ribonu-
Treatment with Na-cyclamate (Na-c) significantly changes cleotide (3 mM). All thresholds were measured twice in all
human taste sensitivity for the monosaccharides glucose subjects. We found the additions of inosine monophosphate
(pyranose) and fructose (furanose) and the structurally (IMP), guanosine monophosphate (GMP), and adenosine
different high-intensity sweeteners Na-c, D-tryptophan monophosphate (AMP), lowered the MPG threshold for
(D-tryp), and sucralose (sucrl), as well as the umami stimulus every subject and were statistically significant p <0.001.
Uridine monophosphate (UMP) yielded mixed results that sensitivity to taste, no significant increases in taste inten-
were not significant. Interestingly, cytosine monophosphate sity were observed post- vs. pre- exposure (p = .115). These
(CMP) raised glutamate thresholds in 6 of 10 subjects, results demonstrate that performing a difficult chemosen-
suggesting it is an inhibitor of glutamate taste in humans. sory task results in a transient increase in taste sensitivity,
The average detection threshold of MPG was 1.75−03 and which we suggest reflects a temporary effect of top- down
with the addition of IMP, the most robust enhancer, the modulation on increasing the efficiency of central taste
threshold was decreased by approximately two orders of processing. Acknowledgements: Supported by NIDCD
magnitude to 4.46−05. CMP increased the MPG threshold grant R01 DC006706
by approximately ten fold in the subject for whom it was
most inhibitory. The rank order of effect on increasing sen-
sitivity to glutamate was IMP > GMP> AMP >> UMP #P247 POSTER SESSION V: HUMAN TASTE
// CMP. We will explore associations among perceptual PSYCHOPHYSICS; OLFACTION RECEPTORS;
differences in synergistic effects of various ribonucleo- TASTE DEVELOPMENT
tides with genetic variations in individual oral glutamate
receptors. Acknowledgements: Funded in part by NIH DC The Role of Glutamate in Infant Satiation: a Model System
02995 to PASB. Alison K. Ventura1, 2, Loma B. Inamdar2, Julie A. Mennella2
1
Drexel University Philadelphia, PA, USA

2
PSYCHOPHYSICS; OLFACTION RECEPTORS; We recently discovered that human infants consumed less
TASTE DEVELOPMENT to satiation when feeding formulas high in free glutamate
(extensively protein hydrolysate formulas; ePHF) than
Transient top-down modulation of gustatory sensitivity when consuming a formula low in free glutamate, such as
in humans standard cows’ milk formulas (CMF). The purpose of this
Maria G Veldhuizen1,2, Elias D Lustbader1, Dana M Small1,2,3 study was to use this model system to characterize the tim-
ing and patterning of cues infants use to signal satiation.
THE JOHN B PIERCE LABORATORY NEW HAVEN, CT, USA,
1
In this within-subjects study, 41 infants <4 months of age
Department of Psychiatry, Yale School of Medicine NEW
2
were tested on two separate days under infant-led feeding
HAVEN, CT, USA, 3Department of Psychology, Yale University
conditions. In counterbalanced order, infants were vide-
NEW HAVEN, CT, USA
otaped as they fed CMF on one test day and CMF with
Perceptual systems are dynamic. Sensitivity may decrease added free glutamate (CMF+glu) comparable to levels
due to adaptation or increase as a result of directed found in ePHF on the other. Raters blinded to the con-
attention. In three studies we tested the hypothesis that ditions coded the timing and frequency of a variety of
sustained directed attention to oral stimulation would behavioral cues and determined the duration of feeding.
influence gustatory sensitivity. In study one, six subjects The infants consumed significantly less formula to satia-
rated the intensity of taste stimuli (sucrose, sucralose, cit- tion and tended to spend less time feeding CMF+glu than
ric acid, sodium chloride, monosodium glutamate) three CMF. Preliminary analyses revealed that while there were
times before and after they performed a triangle task no differences in the types and frequencies of behaviors
requiring sustained directed attention to oral sensation. In exhibited between feeds, infants displayed satiety behav-
the triangle test subjects were presented with two quali- iors earlier when feeding the CMF+glu formula compared
tatively similar flavor stimuli and asked to “pick the odd to the CMF. In sum, we have identified a model system to
one out”. They repeated this task 12 times and then rated experimentally manipulate and study infant satiation, an
the intensity of the taste stimuli again. Subjects returned area of research that is needed both for practical concerns
to the lab four times and repeated this procedure. As pre- of optimizing infant feeding and for theoretical concerns
dicted, taste stimuli were rated as more intense post- vs. focusing on understanding mechanisms underlying hun-
pre- triangle test (p=.009). However, the effect was tran- ger, satiation and satiety. Acknowledgements: Supported
sient in that taste stimuli were rated as less intense during by Grants R01HD37119 and HD072307 from the Eunice
the pre-triangle test vs. the previous days’ post- triangle Kennedy Shriver National Institute of Child Health and
test (p=.000). In study two we replicated these findings Human Development, a National Research Service Award
with a sample of 9 subjects (p=.000 and p=.037). In study F32HD063343 from the Eunice Kennedy Shriver National
three we tested whether consumption of the flavor stimuli Institute of Child Health and Human Development, and
without the performance of the triangle test would result an investigator-initiated grant from Ajinomoto, Inc. The
in sensitivity changes. Consistent with the hypothesis that content is solely the responsibility of the authors and does
it is the performance of the discrimination task, rather not necessarily represent the official views of the Eunice
than mere exposure to flavors that produces the increased Kennedy Shriver National Institute of Child Health and
Human Development. The funding agencies had no role in #P249 POSTER SESSION V: HUMAN TASTE
the design and conduct of the study in the collection, anal- PSYCHOPHYSICS; OLFACTION RECEPTORS;
ysis, and interpretation of the data or in the preparation, or TASTE DEVELOPMENT
review of the abstract.
Re-Engineering of Olfactory Receptor OlfCc1 Toward
Directed Ligand Selectivity
#P248 POSTER SESSION V: HUMAN TASTE Allison P Berke1, Francine Acher2, Hugues O. Bertrand3, John
PSYCHOPHYSICS; OLFACTION RECEPTORS; Ngai4
TASTE DEVELOPMENT 1
Joint Graduate Group in Bioengineering, University of
Identification of Neurogenetic Networks that Contribute California Berkeley, CA, USA, 2Laboratoire de Chimie et
to Natural Variation in Olfactory Behavior in Drosophila Biochimie Pharmacologiques et Toxicologiques, Unite
melanogaster Mixte de Recherche 8601, Centre National de la Recherche
Scientifique, Universite Rene Descartes-Paris V Paris, France,
Robert R H Anholt1,2,3, Shilpa Swarup2,3, Wen Huang2,3, Trudy 3
Accelrys Orsay, France, 4Department of Molecular and Cell
F C Mackay2,3
Biology and Helen Wills Neuroscience Institute Berkeley, CA,
1
North Carolina State University/Biology Raleigh, NC, USA, USA
2
North Carolina State University/Genetics Raleigh, NC, Fish sense food cues in their aqueous environment using fam-
USA, 3North Carolina State University/WM Keck Center for ily C GPCR olfactory receptors. These C family receptors are
Behavioral Biology Raleigh, NC, USA characterized by a large N-terminal “Venus flytrap” domain.
Despite major advances in the identification and char- Zebrafish olfactory receptor OlfCc1 is a broadly-expressed
acterization of odorant receptors, little remains known ortholog of mammalian V2R2, which shares sequence simi-
about the genetic basis of individual variation in olfac- larity with the human Calcium-sensing Receptor (CaSR). C
tory perception. This issue can be addressed in Drosophila family receptors are predicted to respond to amino acids, as
melanogaster, where olfactory behavior can be easily quan- do the previously identified OlfCa1 and CaSR. The expres-
tified for large numbers of genetically identical individu- sion of OlfCc1 in the entire microvillous olfactory neuron
als reared under controlled conditions. We used 168 fully population in the zebrafish, as well as its sequence homology
sequenced, inbred, wild-derived lines from the Drosophila with CaSR, make it an interesting target for de-orphaning
melanogaster Genetic Reference Panel (DGRP) to perform and engineering, as it could play a generalized behavio-
three complementary genome wide association (GWA) ral role in zebrafish chemosensation. In silico modeling of
studies to identify alleles that affect natural variation in OlfCc1 identified the receptor’s binding pocket and residues
olfactory behavior to benzaldehyde. First, we performed likely to be directly involved in ligand binding. OlfCc1 was
a GWA on DGRP lines to identify top SNPs associated then cloned into a CMVI FLAG-tagged expression vector
with mean differences in olfactory behavior. Second, we and expressed in HEK293 cells. Calcium imaging performed
intercrossed two divergent DGRP lines and constructed on these cells using Fluo-4 calcium-sensitive dye revealed the
an outbred advanced intercross line population, on which calcium-dependent binding profile of OlfCc1, which includes
we performed an extreme quantitative trait locus GWA. amino acids. Amino acid point mutations were then intro-
Finally, we performed a GWA for SNPs affecting the vari- duced to OlfCc1, with the aim of broadening the receptor’s
ance among lines as a one-dimensional screen for inter- binding specificity. These mutations succeeded in altering
acting loci, since there is evidence for epistasis between the sensitivity and specificity of OlfCc1 in accordance with
mutations affecting olfactory behavior and mutational predictions. In conclusion, the binding specificity and sensi-
effects are suppressed in DGRP line genetic backgrounds. tivity of OlfCc1 can be selectively engineered. Additionally,
We find that different elements of the genetic architecture the combination of in silico homology modeling and calcium
that underlies natural variation in olfactory behavior are imaging that constitute this method can be applied to other
revealed in the three GWA analyses, but they converge C family GPCRs, to directly engineer ligand binding capa-
on similar cellular processes and could be functionally bility. Acknowledgements: NSF GRFP and the NIH
validated at a high rate through analysis of mutants. Our
results indicate that polymorphisms that contribute to nat- #P250 POSTER SESSION V: HUMAN TASTE
ural variation in olfactory perception are not restricted to PSYCHOPHYSICS; OLFACTION RECEPTORS;
the chemosensory sub-genome, but that variation in neural TASTE DEVELOPMENT
connectivity and synaptic signaling in the central nervous
Interactions at the olfactory receptor level contribute to the
system is a major determinant of individual variation in
coding of odorant mixtures
odor perception. Acknowledgements: This work was sup-
ported by National Institutes of Health grants GM59469 fouzia El Mountassir1, christine Belloir1, loic Briand1, thierry
and GM45146. Thomas Danguin1, anne marie Le BON1
1
Centre des Sciences du Gout et de l’Alimentation DIJON, contains binding sites for the transcriptional repressor
France, 3 zbtb7b, which is abundantly expressed in the peripheral
Numerous studies reported that the perceptual characteris- zebrafish olfactory system. Preliminary results suggest that
tics of odorant mixtures are often different from those of negative regulation confers a higher specificity of transgene
their individual compounds; e.g. the mixture intensity can expression to ciliated OSNs. A major distinction within the
be higher or lower than the arithmetic sum of each compo- OR repertoire can be made between evolutionary ancient
nent’s intensity. These findings raise the question how odor- class I ORs and the group of class II receptors, which mas-
ants in mixtures are detected and encoded at the peripheral sively expanded in the tetrapod lineage. OR101-1 appears to
level of the olfactory system. We investigated this question be the only class II OR in zebrafish. OSNs that fail to express
through the measurement of human olfactory receptor (OR) a functional OR usually undergo a second OR gene choice
responses to two specific binary mixtures of aldehydes: (i) resulting in heterogeneity of OR expression and widespread
octanal and citronellal, known to induce a configural percep- axonal projection of transgenic OSN axons to olfactory bulb
tion in rats (Kay et al., 2003) and masking effects in humans glomeruli. Transgenic OSNs expressing a deletion allele of
(Burseg et al., 2009); (ii) octanal and methional, known to the OR101-1 gene in contrast target a single specific glomer-
induce masking effects in humans (Burseg et al., 2009). We ulus, in the zebrafish olfactory bulb suggesting that second
used a heterologous expression system (HEK293T cells) OR gene choice is severely restricted in those OSNs. The
in which OR (OR1G1, OR52D1, OR2W1 and OR1A1) results imply that distinct regulatory mechanisms evolved
were transfected transiently. Responses of OR to odorants concomitantly with the appearance of the first class II OR
applied alone or in mixtures were measured by calcium in the teleost lineage. Acknowledgements: TÜBITAK, The
imaging. The results showed various interactions at the Scientific and Technological Research Council of Turkey,
OR level. When octanal was mixed with citronellal, the OR Grant Awards: 107T760, 112T168
response intensity was reduced thus showing subtraction,
compromise or partial addition, depending on the OR and
the concentrations of odorants. Interestingly, the mixture of #P252 POSTER SESSION V: HUMAN TASTE
octanal and methional was found to induce mostly synergy, PSYCHOPHYSICS; OLFACTION RECEPTORS;
whatever the OR. These data strengthen the hypothesis that TASTE DEVELOPMENT
interactions can occur at the OR level and could therefore Olfactory Receptor (OR) switching is influenced by genome
contribute significantly to the olfactory coding of odorant position in olfactory-placode (OP)-derived cells.
mixtures. Acknowledgements: This work is funded by the
National Institute of Agricultural Research and the region Robert P. Lane, Seda Kilinc
of Burgundy Wesleyan University Middletown, CT, USA
We previously characterized olfactory receptor (OR) expres-
#P251 POSTER SESSION V: HUMAN TASTE sion in the OP6 and OP27 cell lines and made two general
PSYCHOPHYSICS; OLFACTION RECEPTORS; observations: OR choice is not a heritable property; and
TASTE DEVELOPMENT the range of OR representation in OP populations appears
biased for a subset of the full OR repertoire. We used cus-
Class-specific regulation of a zebrafish olfactory tom arrays and deep sequencing to analyze the complete
receptor gene expressed OR repertoire in OP cultures. OP6 and OP27 cell
Stefan H. Fuss, Xalid Bayramli, Nuray Sögünmez lines have significant overlap in OR representation, consist-
ent with similar pre-specification of the two founder cells,
Bogazici University, Molecular Biology and Genetics
which had been isolated from a common developmental
Istanbul, Turkey
milieu. However, OR representation in OP cultures is not
Olfactory sensory neurons (OSNs) typically express a single constrained by presumptive expression zones within mouse
allele of a single olfactory receptor (OR) gene from a much olfactory epithelium, as might be predicted if the range of
larger genomic repertoire; a phenomenon that is not well OR choices had been pre- specified by developmental (i.e.,
understood. Experimental evidence suggests that OR expres- spatial) cues. Instead, we find strong evidence for ‘position-
sion is controlled by a combination of long- and short-range effects’: neighboring OR genes are significantly over-repre-
regulatory sequences. We used promoter bashing in trans- sented in divergent populations, suggesting a tendency to
genic zebrafish to identify proximal regulatory sites within switch within versus across OR clusters. Surprisingly, we do
the OR101-1 gene promoter. Positive regulatory sites are not observe differences in common epigenetic marks between
located within the first 500 bp upstream of the transcription “active” versus “inactive” ORs, nor does locus positioning
start site, while more distant sequences confer a repressive relative to nuclear chromocenters appear to be predictive of
effect on transgene expression, even in the presence of strong selection probability. We suggest a switching model in which
genomic enhancers. Interestingly, the repressive sequence the epigenetic microenvironment established by previous
OR transcription increases the likelihood of selection/ re- OR function. Acknowledgements: This work was sup-
selection within that genome region. We hypothesize that ported by the National Institute of Allergy and Infectious
the OP6 and OP27 founder cells had been similarly speci- Disease [AI056402] to L.J.Z and the National Institute on
fied, initially leading to a similar bias for OR switching, but Deafness and Other Communication Disorders (NIDCD)
that in the absence of further developmental cues during [DC001655] to B.W.A. at the National Institutes of Health,
subsequent culturing, probabilistic switching histories have and the Foundation for the National Institutes of Health
resulted in slow evolution from initial biases, thus indepen- through the Grand Challenges in Global Health Initiative
dently evolving OR subrepertoires. Acknowledgements: [VCTR121] to L.J.Z. G.M.P was supported by the NIDCD
NIH R01-DC006267 NSF 0842868 through an NRSA F31 [DC011989].

TASTE DEVELOPMENT
TASTE DEVELOPMENT
Blockade of Insect Odorant Receptor Currents by Amiloride
Functional Analysis Of Nematode GPCRS In Yeast
Derivatives
Muhammad Tehseen, Alisha Anderson, Mira Dumancic,
Gregory M Pask1, Yuriy V Bobkov2, Elizabeth A Corey2, Barry
Lyndall Briggs, Stephen Trowell
W Ache2,3, Laurence J Zwiebel1,4
CSIRO Food Futures Flagship and Division of Ecosystem
1
Department of Biological Sciences, Vanderbilt University
Sciences, Black Mountain Laboratories Canberra, Australia
Nashville, TN, USA, 2Whitney Laboratory, Center for Smell
and Taste, and McKnight Brain Institute, University of The yeast Saccharomyces cerevisiae has been used exten-
Florida Gainesville, FL, USA, 3Departments of Biology and sively for ligand screening of human G-protein coupled
Neuroscience, University of Florida Gainesville, FL, USA, receptors, due to its ease of genetic manipulation, low cost,
4
Department of Pharmacology, Vanderbilt Brain Institute rapid growth, and eukaryotic secretory pathway. Although
and Center for Human Genetics, Institutes of Chemical the Caenorhabditis elegans genome was sequenced 13 years
Biology and Global Health and Program in Developmental ago and encodes over 1,000 GPCRs, of which several hun-
Biology, Vanderbilt University Medical Center Nashville, dred are believed to respond to volatile organic ligands, only
TN, USA one of these receptors, ODR-10, has been linked to a vola-
tile ligand, 2,3-butanedione. ODR-3 is a G-protein a subunit
Insect odorant receptors (ORs) function as heteromeric
believed to be involved in odorant detection and activated by
odorant- gated ion channels consisting of a conserved
ODR-10. Here we report the functional coupling of ODR-
coreceptor, Orco, and an odorant-sensitive tuning subu-
10 to the yeast pheromone signalling pathway using the
nit. Although some OR modulators have been identified,
yeast - C. elegans chimaeric Gα subunit (GPA-1:ODR-3).
an extended library of pharmacological tools is currently
Interactions between ODR-10, ODR-3 and the chimaera
lacking and would aid in furthering our understanding
were confirmed using the split ubiquitin yeast two- hybrid
of insect OR complexes. Using whole-cell patch clamp
system. We also report the tailoring of a Saccharomyces
techniques, we demonstrate that amiloride and several
cerevisiae strain for the analysis of C. elegans chemorecep-
derivatives, which have been extensively used as block-
tor function. In this study, a yeast gpa1D ste2D sst2D far1D
ers for various ion channels and transporters, also block
odorant-gated currents from two OR complexes from the quadruple mutant strain was constructed to efficiently cou-
ple nematode olfactory receptors with the yeast signalling
malaria vector mosquito Anopheles gambiae, AgOr48
pathway. We used two different reporters: green fluorescent
+ AgOrco and AgOr65 + AgOrco. In addition, currents
protein and ß-galactosidase, to verify activation of the sig-
from both heteromeric OR complexes were susceptible
nal transduction pathway by ligand activated GPCR inter-
to HMA blockade when activated by VUAA1, an ago-
actions. With this heterologously engineered yeast system,
nist that targets the Orco channel subunit. HMA was also
we aim to accelerate the de-orphaning of C. elegans GPCR
capable of blocking VUAA1-evoked currents of Orco
proteins.
homomers from 4 different insect orders, demonstrating
that HMA blockade is not unique to AgOR complexes.
Additionally, amiloride derivatives have provided insights #P255 POSTER SESSION V: HUMAN TASTE
into the properties of both the channel pore and spon-
taneous gating across different insect OR complexes.
TASTE DEVELOPMENT
Amiloride derivatives therefore represent a valuable class
of channel blockers that can be used to further investigate Differentiating activation of intracellular signaling
the pharmacological and biophysical properties of insect pathways using calcium dynamics
Kirill Ukhanov1, Yuri Bobkov1, Elizabeth A. Corey1, Barry 1

ChemCom S.A. Brussels, Belgium, 2Hôpital Erasme Brussels,
W. Ache1,2 Belgium
1
University of Florida, Center for Smell and Taste Gainesville, Background: Olfactory recognition is mediated by a large rep-
FL, USA, 2University of Florida, Depts. of Biology and ertoire of olfactory receptors (ORs). The human genome con-
Neuroscience Gainesville, FL, USA tains 851 OR loci. More than 50% of the loci are annotated as
nonfunctional due to frame-disrupting mutations. Furthermore
Mammalian olfactory receptors (ORs) appear to have the
some missense haplotypic alleles can be nonfunctional due to a
capacity to couple to multiple G protein-coupled signal-
substitution of key amino acids governing protein folding or
ing pathways, more specifically phosphoinositide-depend-
interaction with signal transduction components. Beyond their
ent signaling in addition to canonical cAMP-dependent
role in odor recognition, functional ORs are also required for
signaling, in a ligand-selective manner. To better under-
a proper targeting of olfactory neuron axons to their corre-
stand the mechanisms and molecular range of such ligand
sponding glomeruli in the olfactory bulb (Feinstein et al, 2004).
selectivity, we developed a heterologous expression system
Therefore, profiling of OR gene expression in the olfactory epi-
with differential readout of intracellular calcium changes.
thelium provides an opportunity to select frequently expressed
We expressed the mouse eugenol receptor (mOREG) in
and potentially functional ORs for large deorphanization cam-
HEK293T cells together with Gα15 [phospholipase-C
paign. Methods: An AB TaqMan® Low Density Array (TLDA)
(PLC) pathway] and/or Gαolf [adenylate cyclase (AC)
containing probes for 356 predicted OR loci was designed to
pathway], leading to intracellular calcium release or cal-
cium influx through a cyclic nucleotide-gated channel investigate the chemosensory receptor gene expression in olfac-
tory epithelium tissues from 8 individuals. Total RNA isolation,
mutant deficient in Ca-CaM negative feedback, respec-
DNase treatment, RNA integrity evaluation and reverse tran-
tively. Eleven known mOREG agonists were tested,
scription in cDNA were performed for these 8 samples. Then
including eugenol, its analogs, and structurally dissimi-
384 gene targets (including reference genes for normalization)
lar compounds (mousse cristal, nootkatone, orivone).
were analysed using the same RT-qPCR platform. Results: The
PLC-dependent responses differed dynamically [e.g.,
expression of 200 (56%) human OR genes was observed in these
eugenol (τrise = 3.55 ± 0.13 sec; τdecay = 72.42 ± 19.01 sec)
olfactory epithelia, among which 114 were robustly expressed
from AC-dependent responses (τrise = 90.83 ± 9.67 sec;
in all tested individuals. No relation between OR gene expres-
τdecay = 167.15 ± 6.18 sec)], allowing them to be distin-
sion and age or sex was observed. Most of the ORs (>80%)
guished when Gα15 and Gαolf were co-expressed. This
deorphanised at Chemcom or described in the literature were
difference persisted across ligand concentration. All ago-
found in the expressed set.
nists tested activated both pathways [e.g., EC50, eugenol:
76 ± 12 μM (PLC), 78 ± 26 μM (AC)], showing that
mOREG can couple to different G proteins expressed
in the same cell. A larger scale screening is under way
to identify and characterize potential OR-ligand com-
binations that differentially activate downstream signal-
ing pathways. Acknowledgements: NIDCD DC001655,
DC005995

TASTE DEVELOPMENT
Profiling of OR gene expression in the human olfactory
epithelium
Françoise Wilkin1, Christophe Verbeurgt2, Pierre Chatelain1

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Chemical Senses Advance Access published August 1, 2013

Chem. Senses doi:10.1093/chemse/bjt036

Downloaded from http://chemse.oxfordjournals.org/ by Sergey Novikov on October 17, 2013

#20 PLATFORM PRESENTATIONS: OLFACTION Genevieve M. Tauxe, Anandasankar Ray

Olfactory Dysfunction Predicts 5-year Mortality in Department of Entomology, University of California,

promote their continued mitosis following irradiation injury. 1

exposed to daily access to sugar or artificial sweeteners #45 PLATFORM PRESENTATIONS: TASTE

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#51 SYMPOSIUM: EXPERIENCE DRIVEN #53 SYMPOSIUM: EXPERIENCE DRIVEN

University of California, San Diego, CNCB, La Jolla CA

#P35 POSTER SESSION I: MULTIMODAL Department of Pharmacological and Physiological Science

OLFACTION PERIPHERY University of Maryland/Department of Anatomy and

Scaffolding proteins in olfaction Neurobiology Baltimore, MD, USA

#P46 POSTER SESSION I: MULTIMODAL 1

OLFACTION CNS Madison Madison, WI, USA, 2Waisman Center, University of

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infusions do not impair memory for the CS+ but reduce 4

OA leads to direct activation of beta-adrenergic-like OA #P110 POSTER SESSION II: OLFACTION

University of Vermont Burlington, VT, USA, 2Transgenomic

Denver, CO, USA, 2Rocky Mountain Taste and Smell Center

A role for bitter taste receptors in thyroid toxicity and

#P181 POSTER SESSION IV: CHEMICAL Ali Magableh, Robert Lundy

Jean-François Cloutier1,2, Alexandra Brignall1,2, Tyler

Discrimination of homologous alcohol odorants varying in Dustin M Graham, David L Hill

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The NIH Toolbox Brief Gustation Assessment Protocol

being explored in a follow-up experiment by using more USA

Drexel University Philadelphia, PA, USA

#P253 POSTER SESSION V: HUMAN TASTE

Kirill Ukhanov1, Yuri Bobkov1, Elizabeth A. Corey1, Barry 1

#P256 POSTER SESSION V: HUMAN TASTE

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