Brain, Behavior, and Immunity xxx (2015) xxxxxx

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Brain, Behavior, and Immunity

journal homepage: www.elsevier.com/locate/ybrbi

GPR84 deciency reduces microgliosis, but accelerates dendritic

degeneration and cognitive decline in a mouse model of Alzheimers
disease
Julie Audoy-Rmus a, Lusine Bozoyan a, Aline Dumas a, Mohammed Filali a, Cynthia Lecours a,
Steve Lacroix a,b, Serge Rivest a,b, Marie-Eve Tremblay a,b, Luc Vallires a,b,
a
b

Axis of Neuroscience, University Hospital Center of Quebec, Quebec, QC, Canada

Department of Molecular Medicine, Laval University, Quebec, QC, Canada

a r t i c l e

i n f o

Article history:
Received 3 September 2014
Received in revised form 6 January 2015
Accepted 6 January 2015
Available online xxxx
Keywords:
Neuroinammation
Neurodegeneration
Microglial cells
Chemotaxis
b-Amyloid
Multiple sclerosis
Endotoxemia
Lipopolysaccharide
G-protein coupled receptor 84

a b s t r a c t
Microglia surrounds the amyloid plaques that form in the brains of patients with Alzheimers disease
(AD), but their role is controversial. Under inammatory conditions, these cells can express GPR84, an
orphan receptor whose pathophysiological role is unknown. Here, we report that GPR84 is upregulated
in microglia of APP/PS1 transgenic mice, a model of AD. Without GPR84, these mice display both accelerated cognitive decline and a reduced number of microglia, especially in areas surrounding plaques. The
lack of GPR84 affects neither plaque formation nor hippocampal neurogenesis, but promotes dendritic
degeneration. Furthermore, GPR84 does not inuence the clinical progression of other diseases in which
its expression has been reported, i.e., experimental autoimmune encephalomyelitis (EAE) and endotoxic
shock. We conclude that GPR84 plays a benecial role in amyloid pathology by acting as a sensor for a yet
unknown ligand that promotes microglia recruitment, a response affecting dendritic degeneration and
required to prevent further cognitive decline.
2015 Elsevier Inc. All rights reserved.

1. Introduction
The amyloid cascade hypothesis states that altered processing
of amyloid precursor protein lead to the release of b-amyloid peptides (e.g., Ab42), which initiate the pathological process underlying AD (Tanzi and Bertram, 2005). These peptides can aggregate
into soluble oligomers and then into insoluble brils that accumulate to form macroscopic amyloid plaques. Whether all of these
forms are toxic and how they cause toxicity are questions that
are still debated. A prevailing view is that the oligomers, more
toxic than the brils, bind to and alter the function of certain membrane proteins, resulting in synaptic dysfunction, dendritic degeneration and neuronal death (Benilova et al., 2012).
Any brain damage, including that caused by b-amyloid, triggers
activation of microglia, the resident immune cells of the CNS. These
cells cluster around amyloid plaques, in which they extend cyto-

Corresponding author at: Axis of Neuroscience, University Hospital Center of

Quebec, 2705 Laurier Boulevard, Room T2-50, Quebec, QC G1V 4G2, Canada. Tel.: +1
418 525 4444x46233; fax: +1 418 654 2761.
E-mail address: [email protected] (L. Vallires).

plasmic processes (Combs, 2009). The signicance of this response

is controversial, but one possibility is that microglia exert an overall benecial effect, as suggested by behavioral studies showing
that microglia depletion or genetic deletions affecting microglia
accelerate cognitive decline in a model of AD (i.e., the APP/PS1
transgenic mouse) (Simard et al., 2006; Naert and Rivest, 2011;
Song et al., 2011). However, this effect does not seem to be related
to the ability of microglia to eliminate parenchymal amyloid plaques, as the latter were not affected (Grathwohl et al., 2009;
Mildner et al., 2011) or only modestly (Simard et al., 2006;
Tahara et al., 2006; Naert and Rivest, 2011; Song et al., 2011) in
the above paradigms. An opposite possibility is that chronically
activated microglia exert deleterious effects, for example, by
phagocytosing neuronal elements and producing potentially neurotoxic molecules such as TNF (Tan et al., 1999, 2002; Fuhrmann
et al., 2010; Lee et al., 2010; Weitz and Town, 2012). Further studies on the molecular mediators responsible for such effects are
required to clarify the seemingly dichotomous roles of microglia
in AD.
Activated microglia express GPR84, a seven-transmembrane
domain receptor of the rhodopsin superfamily that shows limited

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Please cite this article in press as: Audoy-Rmus, J., et al. GPR84 deciency reduces microgliosis, but accelerates dendritic degeneration and cognitive
decline in a mouse model of Alzheimers disease. Brain Behav. Immun. (2015), http://dx.doi.org/10.1016/j.bbi.2015.01.010

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similarity to other known receptors (Fredriksson et al., 2003; Foord

et al., 2005). In the CNS, GPR84 expression is restricted to microglia
and observed in different pathological conditions, including EAE,
endotoxemia, cuprizone-induced demyelination and axotomy
(Bedard et al., 2007; Bouchard et al., 2007; Gamo et al., 2008). In
the periphery, GPR84 is mainly expressed by cells of the myeloid
lineage, such as monocytes, macrophages and neutrophils
(Youse et al., 2001; Wang et al., 2006; Suzuki et al., 2013). In vitro
studies have revealed that GPR84 signals through a Gi/o pathway in
response to hydroxylated (and to a lesser extent nonhydroxylated)
medium-chain free fatty acids (FFAs) of 914 carbons in length
(Wang et al., 2006; Suzuki et al., 2013). These can induce chemotaxis and amplify lipopolysaccharide (LPS)-induced cytokine production in myeloid cells (Wang et al., 2006; Suzuki et al., 2013).
Nevertheless, the pathophysiological role of GPR84 and the nature
of its endogenous ligand(s) remain unknown.
The goals of the present study were to: (1) examine the expression of GPR84 in APP/PS1 mice; and (2) determine its importance
in this model and two others in which GPR84 expression has been
reported, i.e., EAE and endotoxic shock (Bouchard et al., 2007). Our
results reveal that GPR84 exerts no signicant effect on the progression of the latter two diseases, but promotes microgliosis and
dendritic homeostasis in APP/PS1 mice. Therefore, this study
ascribes for the rst time a role to GPR84 in vivo.

2. Materials and methods

2.1. Animals
GPR84-decient mice were obtained from DeltaOne (Deltagen)
and bred with C57BL/6 J mice (Jackson Laboratory) for at least 5
generations. These mice are reported to be indistinguishable from
wild-type littermates under normal conditions (Venkataraman and
Kuo, 2005). APP/PS1 transgenic mice (Borchelt et al., 1997) (Jackson Laboratory, B6C3-Tg(APP695)3Dbo Tg(PSEN1)5Dbo/J) expressing a chimeric amyloid precursor protein (APPSwe) and human
presenilin 1 (A246E variant) under the control of the mouse prion
protein promoter were bred with GPR84-decient mice for at least
3 generations. The genotypes were conrmed by PCR as recommended by the providers (primers used: GPR84, 50 -acagctcagatgccaacttctcctg-30 , 50 -tcctagagcaatgagacagagggtg-30 and 50 gacgagttcttctgaggggatcgatc-30 ; APP, 50 -gactgaccactcgaccaggttctg30 and 50 -cttgtaagttggattctcatatccg-30 ; PS1, 50 -aatagagaacggcaggagca-30 and 50 -gccatgagggcactaatcat-30 ). The experiments were
performed with the approval of our institutional animal ethics
committee. Sex-matched and age-matched wild-type littermates
were used as controls. EAE and LPS-treated mice were 2 months
of age, whereas APP/PS1 mice were tested at 212 months of age.

2.2. EAE induction and clinical evaluation

Mice were subcutaneously injected into both anks with a total
of 200 ll of emulsion containing 300 lg of myelin oligodendrocyte
glycoprotein (MOG) peptide 3555 (AnaSpec) dissolved in saline
and mixed with an equal volume of complete Freunds adjuvant
containing 500 lg of killed Mycobacterium tuberculosis H37 RA
(Difco Laboratories). The mice were also injected intraperitoneally
with 20 lg/kg of pertussis toxin (List Biological Laboratories)
immediately and 2 days after immunization. The clinical signs of
EAE were scored daily using the following scale: 0, no detectable
sign; 1, tail accidity; 2, hindlimb weakness and poor righting ability; 3, hindlimb paralysis/paresis; 4, hindlimb paralysis and forelimb paraparesis; 5, dead or sacriced for humane reasons.

2.3. LPS treatment and sickness behavior assessment

LPS from Escherichia coli O55:B5 (SigmaAldrich) was dissolved
in pyrogen-free saline and injected intraperitoneally at a dose of
1 mg/kg. At the indicated time points, the mice were placed in
the stem of a T-maze made of clear Plexiglas (stem: 64  12 cm;
arms: 30  12 cm; height of the walls: 16 cm) and divided into
thirteen segments (10  10 cm). The locomotor activity of the mice
was evaluated by counting the number of rearings and segments
crossed during a 4-min session. A segment was counted when
crossed by the whole body, excluding the tail. The T-maze was
cleaned after each session.
2.4. Cognitive analysis
The cognitive performance of APP/PS1 mice was evaluated in a
T-maze (see specications above) lled with water (23 1 C) at a
height of 12 cm and containing a platform (11  11 cm) submerged 1 cm beneath the surface at the end of one or both arms
(Filali and Lalonde, 2009). A trial consisted of placing the mouse
in the stem of the maze and allowing it to swim toward the left
or right arm until it nd a platform. If the platform was not found
after 60 s, the mouse was gently guided toward it. The mouse was
allowed to rest for 20 s on the platform after each trial and for
10 min in its cage after each 10-trial block. During the acquisition
phase, 2 training trials were performed with a platform placed in
both arms, and then a maximum of 48 additional trials were performed without the platform chosen during the second training
trial. The acquisition phase was considered completed when the
mouse passed the criterion of 5 consecutive errorless trials (correct
turn and platform found within 60 s). Two days later, the reversal
phase was conducted using the same protocol, except that the platform was moved to the opposite arm and that there was no training. The number of trials-to-criterion and the time to reach the
platform (escape time) were recorded.
2.5. Immunostaining
Immunohistochemistry was performed using rabbit anti-Iba1
(1:2000, Wako Chemicals), goat anti-doublecortin (DCX) (1:500,
Santa Cruz Biotechnology) and rabbit anti-Ki67 (1:500, Abcam)
antibodies, as described previously (Audoy-Remus et al., 2008).
2.6. In situ hybridization
Brain sections were analyzed for GPR84 mRNA by radioisotopic
in situ hybridization as described previously (Bouchard et al.,
2007).
2.7. Congo red staining
Free-oating sections were stained for 30 min in 0.5% Congo red
diluted in 50% ethanol and differentiated (510 dips) in 0.01%
NaOH diluted in 50% ethanol.
2.8. Stereological analyses
Systematically sampled sections (every 10th section) were
examined in a blinded fashion using a Stereo Investigator system
(Microbrighteld) coupled to a Nikon E800 microscope. To estimate the density of GPR84 mRNA+ cells (in situ hybridization)
and immunostained cells (Iba1, DCX), we used the fractionator or
optical fractionator method, respectively. The areas of interest
were traced and then sampled using the following counting
parameters: distance between counting frames, 200  200 lm
(GPR84 mRNA+ and DCX+ cells) or 600  600 lm (Iba+ cells); frame

Please cite this article in press as: Audoy-Rmus, J., et al. GPR84 deciency reduces microgliosis, but accelerates dendritic degeneration and cognitive
decline in a mouse model of Alzheimers disease. Brain Behav. Immun. (2015), http://dx.doi.org/10.1016/j.bbi.2015.01.010

J. Audoy-Rmus et al. / Brain, Behavior, and Immunity xxx (2015) xxxxxx

size, 600  600 lm (GPR84 mRNA+ cells), 50  50 lm (DCX+ cells)

or 100  100 lm (Iba+ cells); frame and guard zone thickness
(optical fractionator only), 10 lm and P2 lm, respectively. Cells
were counted if their body was at least partially located within
the counting frame and did not touch the exclusion lines. Total
count was converted into cell density taking into account the
counting parameters and section thickness. In one analysis, Iba1+
cells were counted only if the cell body contacted an amyloid plaque. Given its small size, the population of Ki67+ cells was estimated using a modied approach in which all the proles were
counted within the traced areas without using counting frames.
The size of Congo red+ plaques was estimated according the Cavalieri method. Using a 20 Plan Apochromat objective, a point grid of
10  10 lm was overlaid on each section, and the points that fell
within the region of interest were counted. The counts were converted to area estimates taking into account sampling frequency,
magnication, grid size, and section thickness. Plaques counts
were expressed as number per mm2.
2.9. Electron microscopy
Mice were anesthetized and perfused with PBS, followed by
3.5% acrolein (75 ml) and then 4% paraformaldehyde (150 ml) in
100 mM sodium phosphate buffer (pH 7.4). The brains were postxed in 4% paraformaldehyde for 12 h at 4 C and cut at 50 lm
thickness in PBS using a vibratome. Sections containing the
ventral part of the hippocampus or the prefrontal cortex were
postxed in 1% osmium tetroxide, dehydrated in ethanol and
propylene oxide, embedded in Durcupan (Electron Microscopy
Sciences) overnight at room temperature and laminated between
ACLAR lms (Electron Microscopy Sciences) by heating at 55 C
for 4872 h. Areas of the CA1 region, approximately from Bregma
3.07 to 3.51 mm according to the Paxinos and Franklins atlas
(Paxinos and Franklin, 2012), or of the prefrontal cortex, approximately from Bregma +2.93 to +2.45 mm, were excised from the
lms to prepare ultrathin sections (6580 nm). The latter were
examined using a Tecnai G2 Spirit BioTwin electron microscope
(FEI) equipped with an Orca digital camera (Hamamatsu). An
average of 9 sections (60 nm thick) containing the CA1 pyramidal, radiatum and lacunosum-moleculare layers were examined
per mouse. Pyramidal cell bodies showing signs of oxidative
stress were observed, accompanied by degeneration of their
dendrites in the radiatum and lacunosum-moleculare layers. For
quantitative analysis, series of images were randomly taken at
the level of the radiatum, excluding dystrophic tissue with
amyloid deposition, at magnications between 1900 and 4800
to obtain a total surface of >10,000 lm2 per mouse. Proles of
degenerating dendrites were counted and the results were
expressed as number per area. These proles were identied
using a combination of established criteria for dendrites (e.g.,
the presence of synaptic contacts and protuberances such as
spines, lopodia and small branches (Peters et al., 1991)) and
degenerating cellular elements (e.g., condensed cytoplasm,
plasma membrane rufing, mitochondrial alteration and endoplasmic reticulum dilation (Tremblay et al., 2012)). In addition,
approximately 9 sections containing the prefrontal cortex deep
layers were examined in the same manner for the purpose of
qualitative analysis.
2.10. Protein extraction
Hemi-forebrains were collected from saline-perfused mice as
described previously (Lesne et al., 2006). Briey, each was mechanically homogenized by 10 passages through a 20-gauge needle
with 500 ll of buffer (50 mM TrisHCl, pH 7.6, 0.01% NP-40,
150 mM NaCl, 2 mM EDTA, 0.1% SDS, 1 mM phenylmethylsulfonyl

uoride, 1 protease inhibitor cocktail (Sigma)). After centrifugation of the homogenate at 3 000 rpm for 5 min, the supernatant
(soluble, extracellular protein-enriched fraction) was collected
and stored at 80 C until analysis.
2.11. Western blotting
Protein samples (30 lg) were electrophoretically separated in
precast 1020% Tris-Tricine polyacrylamide gels (Bio-Rad Laboratories) and transferred to polyvinylidene diuoride membranes
(PerkinElmer). The latter were boiled for 10 min in PBS, blocked
in Tris-buffered saline containing 0.05% Tween 20 and 5% skim
milk, and incubated in the same buffer supplemented with antiAb42 antibody (clone 6E10, 1/1000, Covance). The membranes
were stained using a horseradish peroxidase-conjugated goat
anti-mouse IgG antibody (1:1000, Jackson ImmunoResearch) and
the Western Lightning Chemiluminescence Reagent Plus kit (PerkinElmer). Membranes were stripped in 25 mM glycineHCl (pH
2.0) containing 1% SDS and stained for b-actin using a mouse
anti-b-actin antibody (MAB1501; 1:5000; Millipore). The integrated density of the bands corresponding to Ab42 monomers
was measured using a gel imaging system (scanner Agfa Arcus II;
NIH ImageJ software, version 1.44o), subtracted from background
and normalized to b-actin.
2.12. Statistical analyses
Data are expressed as mean SE and were compared using oneway or two-way ANOVA (with or without repeated measures) and
post hoc unpaired t tests or Wilcoxon rank-sum tests. Survival
curves were analyzed using the log-rank test. All these tests used
an alpha of 0.05 and were done with JMP software (SAS Institute).
3. Results
3.1. GPR84 is upregulated in microglia of APP/PS1 mice
We have reported that GPR84 is upregulated in microglia under
the control of TNF and IL-1b and during different pathological conditions such as EAE and endotoxemia (Bedard et al., 2007;
Bouchard et al., 2007). To test our assumption that GPR84 would
also be expressed in a neurodegenerative context, we analyzed,
by in situ hybridization, brain sections from APP/PS1 transgenic
mice killed at different ages. This technique was used because reliable immunostaining for GPR84 could not be achieved using commercially available antibodies. Virtually no hybridization signal for
GPR84 mRNA was detected in wild-type mice (data not shown)
and in APP/PS1 mice aged 2 or 3 months (Fig. 1a). In contrast,
strong signals were observed in the hippocampus and cerebral cortex of older transgenic mice, and the number of positive cells
increased with age (Fig. 1be). To conrm the identity of these
cells, adjacent sections were double labeled for GPR84 mRNA and
the microglia-specic marker Iba1 by combined in situ hybridization and immunohistochemistry followed or not by Congo red
staining. We observed that the hybridization signals colocalized
with Iba1 (Fig. 1f) and were generally distributed around or near
Congo red+ amyloid plaques (Fig. 1g). Overall, these results indicate
that microglia overexpress GPR84 chronically in response to bamyloid.
3.2. GPR84 inuences the clinical outcome in APP/PS1 mice, but not in
mice suffering from EAE or endotoxic shock
To assess the importance of GPR84 in neuroimmune processes,
we evaluated the behavioral effects of a targeted deletion of the

Please cite this article in press as: Audoy-Rmus, J., et al. GPR84 deciency reduces microgliosis, but accelerates dendritic degeneration and cognitive
decline in a mouse model of Alzheimers disease. Brain Behav. Immun. (2015), http://dx.doi.org/10.1016/j.bbi.2015.01.010

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Fig. 1. Expression of GPR84 mRNA in the brains of APP/PS1 transgenic mice. (ad), Dark-eld micrographs showing in situ hybridization signals for GPR84 mRNA in the
cerebral cortex (CC) and hippocampus (Hipp.) of APP/PS1 mice killed at the indicated ages. Scale bar = 250 lm. (e), Stereological analysis revealed increased numbers of
GPR84 mRNA+ cells in the brain of APP/PS1 mice compared to wild-types. Signicantly different from the wild types (ANOVA followed by post hoc Students t tests: P < 0.05).
Sample size: 3 per group. (f), Bright-eld image showing hybridization signals (black grains) for GPR84 mRNA at higher magnication in an 8 month-old APP/PS1 mouse. Blue,
thionin counterstaining. Scale bar = 25 lm. (g and h), Micrographs at different magnications showing hybridization signals for GPR84 mRNA (black grains) colocalized with
Iba1 immunostaining (microglia, brown) and located around Congo red-stained amyloid plaques (arrowheads) in cortical sections of a 8 month-old APP/PS1 mouse. Arrows,
examples of double-labeled cells. Scale bars = 25 lm. (For interpretation of the references to colour in this gure legend, the reader is referred to the web version of this
article.)

GPR84 gene in mice: (1) subjected to EAE by immunization with a

MOG peptide; (2) exposed to bacterial LPS; or (3) harboring the APP
and PS1 transgenes. While no intergroup difference was observed
in the severity of EAE (Fig. 2a) and LPS-induced sickness behavior
(Fig. 2b), a greater decline in cognitive performance was observed
in APP/PS1 mice lacking GPR84 compared to APP/PS1 mice expressing GPR84 (Fig. 2c). Indeed, the decline was detectable earlier (i.e.,
at 4 months) and was more severe at least until the age of 8 months.
Thereafter, the decline reached a plateau so that there was no difference anymore after a year. These results were neither attribut-

able to a cognitive problem resulting exclusively from the GPR84

deletion, because the cognitive performance of GPR84-knockout
mice lacking the APP and PS1 transgenes was comparable to that
of wild-type mice, nor were they attributable to a swimmingrelated problem, because all of the mice manifested a similar swimming capacity and motivation to escape from the water during
training sessions (data not shown). Therefore, we conclude that
GPR84 is benecial in a context of b-amyloid-induced toxicity,
but has no apparent clinical impact in the context of EAE and
LPS-induced sickness behavior.

Please cite this article in press as: Audoy-Rmus, J., et al. GPR84 deciency reduces microgliosis, but accelerates dendritic degeneration and cognitive
decline in a mouse model of Alzheimers disease. Brain Behav. Immun. (2015), http://dx.doi.org/10.1016/j.bbi.2015.01.010

J. Audoy-Rmus et al. / Brain, Behavior, and Immunity xxx (2015) xxxxxx

Fig. 2. Clinical assessment of GPR84 knockout and wild-type mice in three disease models. (a) No intergroup difference in disease severity was detected in mice with chronic
EAE induced by immunization with a MOG peptide (left panel, Wilcoxon rank sum tests: P > 0.1; right panel, log-rank test: P = 0.77). Sample size: 54 knockouts and 59 wild
types. (b) No intergroup difference in locomotor activity (an indicator of sickness) was detected in mice injected intraperitoneally with LPS (1 mg/kg), as evaluated by
counting the number of segments crossed and rearings in a T-maze (two-way ANOVA with repeated measures: P = 0.3 (left panel) or 0.2 (right panel)). Sample size: 16
knockouts and 18 wild types. (c) A decit in spatial learning was detected in APP/PS1 transgenic mice lacking GPR84, as indicated by the escape time and number of trials-tocriterion recorded during the reversal learning phase in a water T-maze. The dashed lines indicate the error bars of non-APP/PS1 mice at 4 months of age. Signicantly
different from APP/PS1 mice decient in GPR84 (Wilcoxon post hoc tests: P 6 0.02 (left panel) or 0.04 (right panel)). Sample size: 20 knockouts and 26 wild types.

3.3. GPR84 promotes the accumulation of microglia around amyloid

plaques
Microgliosis is a typical response to neurodegenerative diseases
and has been well documented in mouse models of AD (Combs,
2009). As previously reported in APP/PS1 mice and shown in
Fig. 3a, microglia are recruited to surround amyloid plaques and
extend cytoplasmic processes toward them. To determine whether
GPR84 plays a role in such recruitment, we estimated the number
of ramied microglia immunostained for Iba1 in the cerebral cortex and hippocampus of APP/PS1 mice decient or not in GPR84
using a stereological counting technique. We found reductions
ranging from 16 to 32% in the number of microglia that contacted
amyloid plaques in APP/PS1 mice lacking GPR84 at all ages analyzed (from 4 to 12 months) (Fig. 3b). There was also a small downward trend in the total number of microglia in the cortex and
hippocampus, reaching statistical signicance at some ages
(Fig. 3c). No major difference in microglial morphology was
observed between the groups. These results suggest that GPR84
contributes to the recruitment of microglia around amyloid plaques in the brain of APP/PS1 mice.
3.4. GPR84 does not affect b-amyloid deposition and accumulation
b-Amyloid is responsible for the cognitive decline observed in
APP/PS1 mice (Borchelt et al., 1997). A predominant species is

Ab42, which can aggregate to form insoluble amyloid deposits in

the extracellular compartment (Burdick et al., 1992; Jarrett et al.,
1993). To determine whether GPR84 inuences b-amyloid deposition, we used stereological methods to estimate the number and
size of amyloid plaques stained with Congo red in the cortex and
hippocampus of APP/PS1 mice aged 412 months and expressing
or not GPR84. As shown in Fig. 4ac, we observed age-related
changes in both of these parameters, but no genotype-related
change. To further examine whether GPR84 affects the accumulation of Ab42, which may also be toxic under non-aggregated forms,
we estimated the amount of Ab42 monomers by Western blotting
in extracellular protein-enriched fractions from the forebrain.
Again, the results increased with age, but did not differ with genotype (Fig. 4d and e). Therefore, GPR84 does not seem to inuence
the metabolism of b-amyloid.
3.5. The lack of GPR84 increases b-amyloid-induced dendritic
degeneration
It is known that b-amyloid induces dendritic degeneration in
APP/PS1 mice (Tsai et al., 2004; Dikranian et al., 2012) and AD
brains (Masliah et al., 1994; Scheff et al., 2007), but whether
microglia exert a protective or deleterious role in this phenomenon
is still a matter of speculation (Siskova and Tremblay, 2013). To
assess whether microglial GPR84 inuences dendritic degeneration, we examined the hippocampal CA1 stratum radiatum of

Please cite this article in press as: Audoy-Rmus, J., et al. GPR84 deciency reduces microgliosis, but accelerates dendritic degeneration and cognitive
decline in a mouse model of Alzheimers disease. Brain Behav. Immun. (2015), http://dx.doi.org/10.1016/j.bbi.2015.01.010

J. Audoy-Rmus et al. / Brain, Behavior, and Immunity xxx (2015) xxxxxx

Fig. 3. Microgliosis in APP/PS1 transgenic mice decient or not in GPR84. (a) Cortical section immunostained for Iba1 (microglia, arrowheads) and counterstained with Congo
red (amyloid plaque, arrow). Scale bar = 25 lm. (b) A reduced number of microglia per amyloid plaque was observed in both the hippocampus and cortex of APP/PS1 mice
lacking GPR84. Signicantly different from the wild types (two-way ANOVA: P 6 0.0001 for genotype or <0.0008 for age; post hoc Students t tests: P < 0.05). Sample size: 6
12 per group. (c) Stereological analysis revealed a reduced density of microglia in the hippocampus and cortex of APP/PS1 mice lacking GPR84. Signicantly different from
the wild types (two-way ANOVA: P 6 0.0015 for genotype or <0.0001 for age; post hoc Students t tests: P < 0.05). Sample size: 612 per group. (For interpretation of the
references to colour in this gure legend, the reader is referred to the web version of this article.)

Fig. 4. Amyloid burden in APP/PS1 transgenic mice decient or not in GPR84. (a) Nomarski micrograph of a cortical section stained with Congo red showing an amyloid
plaque (arrow). Scale bar = 25 lm. (b and c) Stereological analyses revealed no difference in the number and area of plaques in the hippocampus and cortex of APP/PS1 mice
lacking GPR84 (two-way ANOVA: P > 0.05 for genotype or <0.0001 for age). Sample size: 614 per group. (d and e) Western blotting revealed no intergenotypic difference in
the amount of Ab42 monomers in brain extracts enriched for extracellular proteins (two-way ANOVA: P = 0.82 for genotype or <0.0001 for age). Sample size: 49 per group.
(For interpretation of the references to colour in this gure legend, the reader is referred to the web version of this article.)

6 month-old APP/PS1 mice expressing or not GPR84 using electron

microscopy. This dendritic layer was chosen because of its involvement in spatial learning tasks (Tsien et al., 1996), such as the water
T-maze, and because it is the site of synaptic changes that have
been well documented in mouse models of AD (Spires and
Hyman, 2004; Spires-Jones and Knafo, 2012). In addition, it has
recently been reported that dendritic degeneration in this region
is associated with hyperexcitability and neuronal circuit dysfunction in APP/PS1 mice (Siskova et al., 2014). As expected, we
observed many degenerating dendrites in our APP/PS1 mice
(Fig. 5af), but not in aged-matched wild-type controls. Proximal
to distal dendrites showed typical features of degeneration, including a condensed, electron-dense cytoplasm, frequent rufing of the
plasma membrane, dilation of the endoplasmic reticulum and
mitochondrial dysfunction, as previously described during normal
aging and b-amyloid deposition (Oster-Granite et al., 1996;
Tremblay et al., 2012). Similar degenerating elements were
observed in the CA1 stratum lacunosum-moleculare, which contains the distal dendritic trees of the same pyramidal cells, and

in the deep layers of the prefrontal cortex (not shown). Quantitative analysis at the level of the stratum radiatum revealed a 2.5fold increase in the number of degenerating dendritic proles in
APP/PS1 mice lacking GPR84 compared to APP/PS1 mice expressing
this gene (Fig. 5c). This result indicates that the absence of GPR84
in microglia compromises the integrity of neuronal circuits in APP/
PS1 mice.
3.6. The lack of GPR84 does not affect adult hippocampal neurogenesis
Neurogenesis in the adult hippocampal dentate gyrus plays a
role in spatial learning (Zhang et al., 2008; Clelland et al., 2009;
Deng et al., 2010; Arruda-Carvalho et al., 2011) and is affected by
inammation (Vallieres et al., 2002; Ekdahl et al., 2003; Monje
et al., 2003; Belarbi and Rosi, 2013) and transgenic expression of
APP/PS1 (Verret et al., 2007; Demars et al., 2010). To rule out the
possibility that the cognitive decit driven by the loss of GPR84
in APP/PS1 mice is due to impaired neurogenesis, we examined
the proliferation and differentiation of neural progenitors in the

Please cite this article in press as: Audoy-Rmus, J., et al. GPR84 deciency reduces microgliosis, but accelerates dendritic degeneration and cognitive
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J. Audoy-Rmus et al. / Brain, Behavior, and Immunity xxx (2015) xxxxxx

granular layer of the dentate gyrus in APP/PS1 mice expressing or

not GPR84. While a decrease in the number of cells expressing the
proliferation marker Ki67 or the neuroblast marker DCX was
observed with age, no intergenotype difference was detected
(Fig. 6). Therefore, the poorer cognitive performance resulting from
the absence of GPR84 in the APP/PS1 background is not attributable to a defect in adult hippocampal neurogenesis.

4. Discussion
Two main ndings emerge from the present work. First, microglia increase their expression of GPR84 not only upon EAE, endotoxemia and nerve injury, as previously reported (Bedard et al., 2007;
Bouchard et al., 2007; Gamo et al., 2008), but also in a neurodegenerative context. GPR84 can therefore be considered as a general
activation marker for microglia. Second, GPR84 contributes to
b-amyloid-induced microgliosis, a response that is required to promote dendritic homeostasis and prevent further cognitive decline.
As discussed below, these ndings help to understand the role of
GPR84 in vivo and the importance of microglia in AD.
One question that arises is how microglia protects from b-amyloid toxicity? Our results indicate that GPR84 inuences neither
the formation or elimination of b-amyloid plaques, nor the concentration of soluble Ab42, one of the most abundant and toxic form of
b-amyloid. This is in agreement with previous studies showing that
microglia do not affect the plaques (Grathwohl et al., 2009;
Mildner et al., 2011) or at best modestly (Simard et al., 2006;
Tahara et al., 2006; Naert and Rivest, 2011; Song et al., 2011). Furthermore, our results reveal a higher number of degenerating dendrites in the absence of GPR84, leading us to infer that a microglial
response regulated by GPR84 is required to protect dendrites from
further degeneration. Alternatively, it is possible that the absence
of GPR84 reduces the ability of microglia to remove degenerating
dendrites, which may affect neuronal plasticity and thus cognition.
The importance of microglia in neuronal plasticity is increasingly
appreciated (Tremblay and Majewska, 2011; Tremblay et al.,

Fig. 5. Dendritic degeneration in APP/PS1 mice decient or not in GPR84. (af)

Electron micrographs of degenerating dendrites (arrows) in the hippocampal CA1
stratum radiatum of 6 month-old APP/PS1 mice. Note their condensed cytoplasm
and dilated endoplasmic reticulum, which are a typical signs of degeneration,
compared to those of normal dendrites (arrowheads). Abbreviations: er, endoplasmic reticulum; m, mitochondria; s, synapse. Scale bars = 2 lm (a) or 500 nm (bf).
(g) Quantication, using images taken from the stratum radiatum, revealed an
increased number of degenerating dendritic proles in APP/PS1 mice lacking GPR84
compared to APP/PS1 mice expressing this gene (Students t test: P = 0.012). Sample
size: 5 per group.

Fig. 6. Hippocampal neurogenesis in APP/PS1 transgenic mice lacking or not

GPR84. (a and b) Sections showing the granule cell layer (GCL) of the dentate gyrus
immunostained (black) for Ki67, a marker of proliferating cells, or DCX, a marker of
newborn neurons. Counterstaining = methyl green. Scale bar = 25 lm. (c and d)
Stereological data showing no intergenotype difference in the number of Ki67+ or
DCX+ cells. Two-way ANOVA: Ki67, P = 0.59 (genotype), <0.0001 (age) or = 0.52
(interaction); DCX, P = 0.56 (genotype), <0.0001 (age) or = 0.45 (interaction).
Sample size: 69 per group.

Please cite this article in press as: Audoy-Rmus, J., et al. GPR84 deciency reduces microgliosis, but accelerates dendritic degeneration and cognitive
decline in a mouse model of Alzheimers disease. Brain Behav. Immun. (2015), http://dx.doi.org/10.1016/j.bbi.2015.01.010

J. Audoy-Rmus et al. / Brain, Behavior, and Immunity xxx (2015) xxxxxx

2011; Blank and Prinz, 2013; Wake et al., 2013), but further work is
needed to elucidate the mechanism by which they inuence AD
progression.
Another important question is what ligand(s) bind to GPR84
in vivo? Previous studies have identied medium-chain FFAs of 9
to 14 carbons in length and two other organic molecules (6-noctylaminouracil and 3,30 -diindolylmethane) as agonists of
GPR84 in vitro (Wang et al., 2006; Suzuki et al., 2013). The hydroxylated forms of these FFAs were found to be more effective than
the nonhydroxylated ones (Suzuki et al., 2013). Derived from the
diet or the hydrolysis of triglycerides, FFAs serve as an energy
source. They can also be hydroxylated and incorporated into sphingolipids (Hama, 2010). The latter are especially abundant in the
nervous system, in which they represent a major component of
myelin (Dupree and Pomicter, 2010). Sphingolipids play not only
a structural role, but also contribute to cell signaling and adhesion
(Hannun and Obeid, 2008). As lipid dysregulation is an important
feature of AD (Benilova et al., 2012; Walter and van EchtenDeckert, 2013), it is tempting to speculate that FFA metabolites,
generated in response to b-amyloid, could act as danger signals.
GPR84 could play a central role in the ability of microglia to perceive these signals and trigger an appropriate response (e.g., chemotaxis, as supported by Suzuki et al., 2013). This is likely as
there are many examples of lipid molecules that bind to G-protein-coupled receptors on immune cells, including other fatty
acids, arachidonic acid metabolites and lysophospholipids
(Kostenis, 2004). Further work will be required to identify the
endogenous GPR84 ligands and go beyond speculation.
In conclusion, this study: (1) identies for the rst time a pathophysiological role of GPR84; (2) provides an in vivo model in
which to search for endogenous GPR84 ligands; and (3) supports
the concept that microglia exert a modest, but benecial effect in
AD. Future studies on GPR84 may help to clarify the link between
lipid dysregulation and neuroinammation in AD.
Conict of interest
No conict of interest is declared by the authors.
Acknowledgments
This work was supported by grants from the Canadian Institutes
for Health Research (CIHR), the Natural Sciences and Engineering
Research Council of Canada and the Multiple Sclerosis Society of
Canada. L.V. and S.L. received Chercheur-Boursier Snior awards
from the Fonds de recherche du Qubec Sant. JAR was supported
by a CIHR Strategic Training Program Grant in genomics. L.B.
received a Merit Scholarship from the Fonds de recherche du Qubec
Nature et technologies. MET was funded by the Banting Research
Foundation and the Scottish Rite Charitable Foundation of Canada.
We thank Pierrot Tremblay, Julie Pag and Martin Parent for technical assistance.
References
Arruda-Carvalho, M., Sakaguchi, M., Akers, K.G., Josselyn, S.A., Frankland, P.W., 2011.
Posttraining ablation of adult-generated neurons degrades previously acquired
memories. J. Neurosci. 31, 1511315127.
Audoy-Remus, J., Richard, J.F., Soulet, D., Zhou, H., Kubes, P., Vallieres, L., 2008. RodShaped monocytes patrol the brain vasculature and give rise to perivascular
macrophages under the inuence of proinammatory cytokines and
angiopoietin-2. J. Neurosci. 28, 1018710199.
Bedard, A., Tremblay, P., Chernomoretz, A., Vallieres, L., 2007. Identication of genes
preferentially expressed by microglia and upregulated during cuprizoneinduced inammation. Glia 55, 777789.
Belarbi, K., Rosi, S., 2013. Modulation of adult-born neurons in the inamed
hippocampus. Front. Cell. Neurosci. 7, 145.
Benilova, I., Karran, E., De Strooper, B., 2012. The toxic Abeta oligomer and
Alzheimers disease: an emperor in need of clothes. Nat. Neurosci. 15, 349357.

Blank, T., Prinz, M., 2013. Microglia as modulators of cognition and neuropsychiatric
disorders. Glia 61, 6270.
Borchelt, D.R., Ratovitski, T., van Lare, J., Lee, M.K., Gonzales, V., Jenkins, N.A.,
Copeland, N.G., Price, D.L., Sisodia, S.S., 1997. Accelerated amyloid deposition in
the brains of transgenic mice coexpressing mutant presenilin 1 and amyloid
precursor proteins. Neuron 19, 939945.
Bouchard, C., Page, J., Bedard, A., Tremblay, P., Vallieres, L., 2007. G protein-coupled
receptor 84, a microglia-associated protein expressed in neuroinammatory
conditions. Glia 55, 790800.
Burdick, D., Soreghan, B., Kwon, M., Kosmoski, J., Knauer, M., Henschen, A., Yates, J.,
Cotman, C., Glabe, C., 1992. Assembly and aggregation properties of synthetic
Alzheimers A4/beta amyloid peptide analogs. J. Biol. Chem. 267, 546554.
Clelland, C.D., Choi, M., Romberg, C., Clemenson, G.D.J., Fragniere, A., Tyers, P.,
Jessberger, S., Saksida, L.M., Barker, R.A., Gage, F.H., Bussey, T.J., 2009. A
functional role for adult hippocampal neurogenesis in spatial pattern
separation. Science 325, 210213.
Combs, C.K., 2009. Inammation and microglia actions in Alzheimers disease. J.
Neuroimmune Pharmacol. 4, 380388.
Demars, M., Hu, Y.S., Gadadhar, A., Lazarov, O., 2010. Impaired neurogenesis is an
early event in the etiology of familial Alzheimers disease in transgenic mice. J.
Neurosci. Res. 88, 21032117.
Deng, W., Aimone, J.B., Gage, F.H., 2010. New neurons and new memories: how does
adult hippocampal neurogenesis affect learning and memory? Nat. Rev.
Neurosci. 11, 339350.
Dikranian, K., Kim, J., Stewart, F.R., Levy, M.A., Holtzman, D.M., 2012. Ultrastructural
studies in APP/PS1 mice expressing human ApoE isoforms: implications for
Alzheimers disease. Int. J. Clin. Exp. Pathol. 5, 482495.
Dupree, J.L., Pomicter, A.D., 2010. Myelin, DIGs, and membrane rafts in the central
nervous system. Prostaglandins Other Lipid Mediat. 91, 118129.
Ekdahl, C.T., Claasen, J.H., Bonde, S., Kokaia, Z., Lindvall, O., 2003. Inammation is
detrimental for neurogenesis in adult brain. Proc. Natl. Acad. Sci. U.S.A. 100,
1363213637.
Filali, M., Lalonde, R., 2009. Age-related cognitive decline and nesting behavior in an
APPswe/PS1 bigenic model of Alzheimers disease. Brain Res 1292, 9399.
Foord, S.M., Bonner, T.I., Neubig, R.R., Rosser, E.M., Pin, J.P., Davenport, A.P.,
Spedding, M., Harmar, A.J., 2005. International Union of Pharmacology. XLVI G
protein-coupled receptor list. Pharmacol. Rev. 57, 279288.
Fredriksson, R., Lagerstrom, M.C., Lundin, L.G., Schioth, H.B., 2003. The G-proteincoupled receptors in the human genome form ve main families. Phylogenetic
analysis, paralogon groups, and ngerprints. Mol. Pharmacol. 63, 12561272.
Fuhrmann, M., Bittner, T., Jung, C.K., Burgold, S., Page, R.M., Mitteregger, G., Haass,
C., LaFerla, F.M., Kretzschmar, H., Herms, J., 2010. Microglial Cx3cr1 knockout
prevents neuron loss in a mouse model of Alzheimers disease. Nat. Neurosci.
13, 411413.
Gamo, K., Kiryu-Seo, S., Konishi, H., Aoki, S., Matsushima, K., Wada, K., Kiyama, H.,
2008. G-protein-coupled receptor screen reveals a role for chemokine receptor
CCR5 in suppressing microglial neurotoxicity. J. Neurosci. 28, 1198011988.
Grathwohl, S.A., Kalin, R.E., Bolmont, T., Prokop, S., Winkelmann, G., Kaeser, S.A.,
Odenthal, J., Radde, R., Eldh, T., Gandy, S., Aguzzi, A., Staufenbiel, M., Mathews,
P.M., Wolburg, H., Heppner, F.L., Jucker, M., 2009. Formation and maintenance
of Alzheimers disease beta-amyloid plaques in the absence of microglia. Nat.
Neurosci. 12, 13611363.
Hama, H., 2010. Fatty acid 2-Hydroxylation in mammalian sphingolipid biology.
Biochim. Biophys. Acta 1801, 405414.
Hannun, Y.A., Obeid, L.M., 2008. Principles of bioactive lipid signalling: lessons from
sphingolipids. Nat. Rev. Mol. Cell Biol. 9, 139150.
Jarrett, J.T., Berger, E.P., Lansbury, P.T.J., 1993. The carboxy terminus of the beta
amyloid protein is critical for the seeding of amyloid formation: implications for
the pathogenesis of Alzheimers disease. Biochemistry 32, 46934697.
Kostenis, E., 2004. A glance at G-protein-coupled receptors for lipid mediators: a
growing receptor family with remarkably diverse ligands. Pharmacol. Ther. 102,
243257.
Lee, S., Varvel, N.H., Konerth, M.E., Xu, G., Cardona, A.E., Ransohoff, R.M., Lamb, B.T.,
2010. CX3CR1 deciency alters microglial activation and reduces beta-amyloid
deposition in two Alzheimers disease mouse models. Am. J. Pathol. 177, 2549
2562.
Lesne, S., Koh, M.T., Kotilinek, L., Kayed, R., Glabe, C.G., Yang, A., Gallagher, M., Ashe,
K.H., 2006. A specic amyloid-beta protein assembly in the brain impairs
memory. Nature 440, 352357.
Masliah, E., Mallory, M., Hansen, L., DeTeresa, R., Alford, M., Terry, R., 1994. Synaptic
and neuritic alterations during the progression of Alzheimers disease. Neurosci.
Lett. 174, 6772.
Mildner, A., Schlevogt, B., Kierdorf, K., Bottcher, C., Erny, D., Kummer, M.P., Quinn,
M., Bruck, W., Bechmann, I., Heneka, M.T., Priller, J., Prinz, M., 2011. Distinct and
non-redundant roles of microglia and myeloid subsets in mouse models of
Alzheimers disease. J. Neurosci. 31, 1115911171.
Monje, M.L., Toda, H., Palmer, T.D., 2003. Inammatory blockade restores adult
hippocampal neurogenesis. Science 302, 17601765.
Naert, G., Rivest, S., 2011. CC chemokine receptor 2 deciency aggravates cognitive
impairments and amyloid pathology in a transgenic mouse model of
Alzheimers disease. J. Neurosci. 31, 62086220.
Oster-Granite, M.L., McPhie, D.L., Greenan, J., Neve, R.L., 1996. Age-dependent
neuronal and synaptic degeneration in mice transgenic for the C terminus of the
amyloid precursor protein. J. Neurosci. 16, 67326741.
Paxinos, G., Franklin, K.B.J., 2012. Paxinos and Franklins the Mouse Brain in
Stereotaxic Coordinates, fourth ed. Academic Press, Orlando.

Please cite this article in press as: Audoy-Rmus, J., et al. GPR84 deciency reduces microgliosis, but accelerates dendritic degeneration and cognitive
decline in a mouse model of Alzheimers disease. Brain Behav. Immun. (2015), http://dx.doi.org/10.1016/j.bbi.2015.01.010

J. Audoy-Rmus et al. / Brain, Behavior, and Immunity xxx (2015) xxxxxx

Peters, A., Palay, S.L., Webster, H.D., 1991. Fine structure of the nervous system:
neurons and their supporting cells. Oxford University Press, New York.
Scheff, S.W., Price, D.A., Schmitt, F.A., DeKosky, S.T., Mufson, E.J., 2007. Synaptic
alterations in CA1 in mild Alzheimer disease and mild cognitive impairment.
Neurology 68, 15011508.
Simard, A.R., Soulet, D., Gowing, G., Julien, J.P., Rivest, S., 2006. Bone marrowderived microglia play a critical role in restricting senile plaque formation in
Alzheimers disease. Neuron 49, 489502.
Siskova, Z., Justus, D., Kaneko, H., Friedrichs, D., Henneberg, N., Beutel, T., Pitsch, J.,
Schoch, S., Becker, A., von der Kammer, H., Remy, S., 2014. Dendritic structural
degeneration is functionally linked to cellular hyperexcitability in a mouse
model of Alzheimers disease. Neuron 84, 10231033.
Siskova, Z., Tremblay, M.E., 2013. Microglia and synapse: interactions in health and
neurodegeneration. Neural. Plast. 2013, 425845.
Song, M., Jin, J., Lim, J.E., Kou, J., Pattanayak, A., Rehman, J.A., Kim, H.D., Tahara, K.,
Lalonde, R., Fukuchi, K., 2011. TLR4 mutation reduces microglial activation,
increases Abeta deposits and exacerbates cognitive decits in a mouse model of
Alzheimers disease. J. Neuroinamm. 8, 92.
Spires-Jones, T., Knafo, S., 2012. Spines, plasticity, and cognition in Alzheimers
model mice. Neural Plast. 2012, 319836.
Spires, T.L., Hyman, B.T., 2004. Neuronal structure is altered by amyloid plaques.
Rev. Neurosci. 15, 267278.
Suzuki, M., Takaishi, S., Nagasaki, M., Onozawa, Y., Iino, I., Maeda, H., Komai, T., Oda,
T., 2013. Medium-chain fatty acid-sensing receptor, GPR84, is a
proinammatory receptor. J. Biol. Chem. 288, 1068410691.
Tahara, K., Kim, H.D., Jin, J.J., Maxwell, J.A., Li, L., Fukuchi, K., 2006. Role of toll-like
receptor signalling in Abeta uptake and clearance. Brain 129, 30063019.
Tan, J., Town, T., Crawford, F., Mori, T., DelleDonne, A., Crescentini, R., Obregon, D.,
Flavell, R.A., Mullan, M.J., 2002. Role of CD40 ligand in amyloidosis in transgenic
Alzheimers mice. Nat. Neurosci. 5, 12881293.
Tan, J., Town, T., Paris, D., Mori, T., Suo, Z., Crawford, F., Mattson, M.P., Flavell, R.A.,
Mullan, M., 1999. Microglial activation resulting from CD40-CD40L interaction
after beta-amyloid stimulation. Science 286, 23522355.
Tanzi, R.E., Bertram, L., 2005. Twenty years of the Alzheimers disease amyloid
hypothesis: a genetic perspective. Cell 120, 545555.

Tremblay, M.E., Majewska, A.K., 2011. A role for microglia in synaptic plasticity?
Commun. Integr. Biol. 4, 220222.
Tremblay, M.E., Stevens, B., Sierra, A., Wake, H., Bessis, A., Nimmerjahn, A., 2011. The
role of microglia in the healthy brain. J. Neurosci. 31, 1606416069.
Tremblay, M.E., Zettel, M.L., Ison, J.R., Allen, P.D., Majewska, A.K., 2012. Effects of
aging and sensory loss on glial cells in mouse visual and auditory cortices. Glia
60, 541558.
Tsai, J., Grutzendler, J., Duff, K., Gan, W.B., 2004. Fibrillar amyloid deposition leads to
local synaptic abnormalities and breakage of neuronal branches. Nat. Neurosci.
7, 11811183.
Tsien, J.Z., Huerta, P.T., Tonegawa, S., 1996. The essential role of hippocampal CA1
NMDA receptor-dependent synaptic plasticity in spatial memory. Cell 87,
13271338.
Vallieres, L., Campbell, I.L., Gage, F.H., Sawchenko, P.E., 2002. Reduced hippocampal
neurogenesis in adult transgenic mice with chronic astrocytic production of
interleukin-6. J. Neurosci. 22, 486492.
Venkataraman, C., Kuo, F., 2005. The G-protein coupled receptor, GPR84 regulates IL4 production by T lymphocytes in response to CD3 crosslinking. Immunol. Lett.
Verret, L., Jankowsky, J.L., Xu, G.M., Borchelt, D.R., Rampon, C., 2007. Alzheimerstype amyloidosis in transgenic mice impairs survival of newborn neurons
derived from adult hippocampal neurogenesis. J. Neurosci. 27, 67716780.
Wake, H., Moorhouse, A.J., Miyamoto, A., Nabekura, J., 2013. Microglia: actively
surveying and shaping neuronal circuit structure and function. Trends Neurosci.
36, 209217.
Walter, J., van Echten-Deckert, G., 2013. Cross-talk of membrane lipids and
Alzheimer-related proteins. Mol. Neurodegener. 8, 34.
Wang, J., Wu, X., Simonavicius, N., Tian, H., Ling, L., 2006. Medium chain fatty acids
as ligands for orphan G protein-coupled receptor GPR84. J. Biol. Chem.
Weitz, T.M., Town, T., 2012. Microglia in Alzheimers disease: its all about context.
Int. J. Alzheimers Dis. 2012, 314185.
Youse, S., Cooper, P.R., Potter, S.L., Mueck, B., Jarai, G., 2001. Cloning and expression
analysis of a novel G-protein-coupled receptor selectively expressed on
granulocytes. J. Leukoc. Biol. 69, 10451052.
Zhang, C.L., Zou, Y., He, W., Gage, F.H., Evans, R.M., 2008. A role for adult TLXpositive neural stem cells in learning and behaviour. Nature 451, 10041007.

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