Effects of Sleep Deprivation on Spatial Memory and Neural

Functioning

Daniel R. Atwood
PDBIO 325 Tissue Biology
Fall 2009

Abstract
Sleep deprivation has a wide range of negative effects on human performance. One of
these effects that has been under a recent widespread investigation is how sleep deprivation
effects spatial memory. Many studies have shown a close relationship between spatial memory
and neurons: those of which are mostly found within the hippocampus. Not only does sleep
deprivation effect the current neurons that have already been proliferated for quite some time,
but it also effects the normal functions of neurogenesis: from survival rate to ability to specialize
and show a neural phenotype. While disruption of sleep for a period shorter than 24-hours has
little effect on both proliferated neurons and neurogenesis alike, prolonged disruption shows
major histological changes which could explain the compromising of spatial memory.

Introduction
Only within the past decade or two, several in-depth investigations have looked into the
complexities of transformations that occur within the brain at both mesoscopic and microscopic
level during the time that which all human kind undergoes on average of 6-8hrs a day: the same
hours spent while placing oneself carefully upon their bed to experience a natural suspension of
consciousness during which the body (and mind) is rejuvenated. Because these studies are fairly
recent, the majority of our society lives in ignorance of its grand importance. Perhaps that is why
most have little regard for sleep, merely putting up with the fact that mammalians are required to
sleep, that perhaps it’s an ‘illness’ that we can one day cure ourselves from. This train of thought,
when combined with the high demands of school and work in today’s world, has turned several
to having severe sleep disorders originating from short and infrequent periods of sleep
deprivation. Besides the recent findings of changes that happen at a histological level within the
human body, many effects of sleep deprivation on human performance have been well known to
the general public for quite some time now. These include irritability, weakened immune system,
and of course, cognitive impartment (Boonstra et al, 2007). However, thanks to many recent
discoveries, many neurological transformations have been witnessed and can explain reasons for
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the compromising of spatial memory during sleep deprivation.

Proliferated Cortical & Hippocampal Dendrite Spines

There have been studies suggesting that sleep deprivation can alter the dendrite portion of
already proliferated neurons. The dendrite, being
the receiving part of the neuron, is a dynamic
structure that can change in response to the
surrounding environment or stimuli. An example
of this was produced by the research carried out
by Neurner-Jehle and his assistants. They
reported a 42% average decrease of cortical
concentration of the dendritic protein, dendrin,
after having several lab rats experience a 24-hour
sleep deprivation (Neuner-Jehle et al, 1996).
In a more recent study, J.-R Chen and his
assistants had similar results. Through the use of
intracellular dye injection, they were able to
reveal a comparison of the changes of dendritic
arbors and dendritic spines between a control Fig. 1 Representative intracellular dye-filled corticospinal
neurons from control (A) and 5-day fatigue rats (B). The
group of rats that were well rested versus others
photograph was taken from one of the serial sections, 60-
that were sleep deprived for 5 days total. The µm-thick, of the injected slice in while the injected

densities of dendritic spines on both the distal fluorescence dye was immunohistochemically converted to
non-fading DAB reaction product. (A1-A4, B1-B4) High
and proximal parts of the dendrite were
magnifications of the corresponding dendritic segments
significantly reduced in rats following 5 days of marked in A and B to show that dendritic spines are well-

fatigue by approximately 23%-32% (as seen in labeled. Scale bar=100 µm for A and B and 10 µm for A1-
A4 and B1-B4.
Fig. 1) and it took about 3-days of rest to restore
the dendritic spine densities on the corticospinal neurons for the 5-day-fatigue rats to match that
of the well rested control group (Chen et al, 2009).

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 Moreover, in the same experiment that J.-R Chen and his assistants conducted, they also
checked the output neurons of CA1 and CA3 hippocampal areas. These two areas are known to
be closely associated with learning and spatial
memory. Just like with the corticospinal
neurons mentioned above, they used the same
intracellular dye to compare the differences in
CA1 and CA3 between the control and the 5-
day sleep deprived group. On the CA1
hippocampal neurons, the distal part of the
apical dendrites were reduced by 31% after 5
days of sleep deprivation, while those on basal
dendrites were reduced even more
dramatically by 52% (as seen in Fig. 2). On
the CA3 hippocampal neurons, spine densities
on proximal apical dendrites were reduced by
20% and on distal apical dendrites by 35%,
while those on basal dendrites were reduced
by 32%. It again took roughly 3-days for the
experimental group to restore their densities of
dendritic spines of all segments found on both
CA1 and CA3 hippocampal neurons to match
Fig. 2 The effect of fatigue on the dendritic spines of those within the control group.
hippocampal CA1 pyramidal neurons. (A) Representative In this experiment, both the reduction of
dendritic segments of the CA1 pyramidal cells of control,
the rats cortical and hippocampal dendrite spines
5-day fatigue and 5-day fatigue followed by 3-day rest rats.
Changes of spine densities were analyzed in B. Number of resulted in reduced spatial memory
basal, proximal apical and distal apical dendritic segments performances. However, after the sleep deprived
analyzed was 31, 24, and 33 for control, 27, 28, and 36 for
experimental group had 3 days of rest, no long-
5 day-fatigue and 36, 47, and 50 for 5-day fatigue and 3-
day rest group. Scaled bar=10 µm. term effects were demonstrated within their
neurological functions.

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 Hippocampal SIRT1/COX Expression & Melatonin
Other than temporary hippocampal dendrite density transformations mentioned above,
these same neurons have been documented to experience another physiological metamorphosis
closely associated with impaired performance in spatial learning and memory. In a recent
experiment, Chang is his assistants used a SIRT1 immunohistochemistry stain to measure the
differences in SIRT1 and COX expression in hippocampal neurons. This time around there four
different experimental rat
groups: a control group that
was subjected to a normal
circadian resting phase; a 5-
day sleep deprived group;
and two additional 5-day
sleep deprived groups with a
dose of melatonin (one group
given 25mg/kg and the other
100mg/kg). After dissection
of the hippocampi, the rats
subjected to 5-day sleep
deprivation without
melatonin dosage
experienced a significant
reduction of hippocampal
SIRT1 and COX Fig. 3 Light photomicrographs showing hippocampal sirtuin (SIRT1) immunoreactivity
expression, both of which in normal untreated (A), total 5-day sleep deprived (TSD) (B), and total 5-day sleep
deprived with different doses of melatonin treated rats (C-F). Note that in normal
were demonstrated by
untreated rats, numerous SIRT1 immunoreactive neurons were observed in hippocampal
impaired performance in pyramidal cell layers (A). However, following 5-days of TSD, the SIRT1
spatial memory (as seen in immunoreactivity was drastically decreased (B). Also note that in rats receiving
different doses of melatonin, the expression of SIRT1 was gradually returned to normal
Fig. 3). This conclusion
levels (C-F). Scale bar = 100 µm.
was further solidified by a
water maze analysis, which revealed that the 5-day sleep deprived rats without melatonin dosage
struggled to successfully complete the maze, taking more time than the other 3 groups. It must

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also be noted that the group of 5-day sleep deprived rats that received doses of melatonin showed
darker SIRT1 immunohistochemistry stains that those of the sleep deprived group without
melatonin intake (Chang et al, 2009). Because of this, melatonin proved to combat the
compromising effects of SIRT1 and COX expressions and therefore leading to a greater spatial
memory performance, similar to the group of rats that were well rested.

N1 Amplitude
Two modern neuroimaging techniques that are commonly used to today to measure and
record neural functions within the brain are functional magnetic resonance imaging (fMRI) and
positron emission tomography (PET). The former measures the blood oxygenation level-
dependent contrast while the latter measures hemodynamic changes by marking blood with a
radioactive tracer (Cabeza & Nyberg, 2000). With respect to sleep deprivation, and through the
use of either of these two neuroimaging techniques, significant alterations in the component N1
have been witnessed within the brain. N1 is a component that is closely associated with the
responsibility of processing visual, auditory, and tactile stimuli that are located in the primary
sensory areas found within the brain. Several experiments have been carried out showing a
pronounced reduction of N1 amplitude in participants that had suffered either a night of sleep
fragmentation or deprivation (Cote et al, 2006). This reduction of N1 amplitude reflected a
decrease of the participants’ response to not only motor stimuli, but also visual and auditory
stimuli. Because of this, spatial memory (that is, the ability to intake new information and recall
it shortly afterwards) was also reduced because of the inability to successfully intake the same
amount of peripheral stimuli that a well-rested individual would be able to process. However,
after having a small dose of caffeine, it has been shown that a sleep-deprived individual can
increase the once reduced N1 amplitude, which means processing of peripheral stimuli can once
again increase (Lorist et al, 1994). It must be noted though, that the substitution of a good night’s
rest with caffeine can only produce the same amplitude of the component of N1 for a short
period of time, usually for only a couple of hours.

Adenosine
Adenosine is a neuromodulator that has been studied quite heavily. In the brain,
adenosine primarily acts as an intercellular messenger that has been shown to be involved in

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processes such as sleep regulation and arousal, as well as inhibition of most other
neurotransmitters (Dunwiddie & Masino, 2001).
Because of adenosine’s inhibition effect of
neurotransmitter’s firing rate, it essentially has the
ability to reduce excitability throughout the entire
brain (as seen in Fig. 4). During sleep deprivation,
adenosine levels have been recorded
accumulating in select parts throughout the brain,
thereby promoting the macroscopic symptom
Fig. 4 Dose-response curve for depression of neuronal is drowsiness (Arrigoni et al, 2006). With the
firing rate by adenosine (n=4–6 trials per dose). The firing
decrease in amplitude of component N1, along
rate was initially set at 2 Hz by injection of a depolarizing
holding current (+50.3±13.3 pA; n=15 neurons). Data are with the accumulation of adenosine, a profound
fit (line) using a sigmoid equation, max compromising effect on spatial memory occurs
inhibition=100±13.7%; IC50=8.8±1.3 μM; and k=2.5±0.3
because of the temporary neurological changes
nM.
that take place.

Neurogenesis
Neurogenesis, the process by which an organism obtains new neurons, was traditionally
believed to occur only during the embryonic stages in the CNS of mammalians. However, many
recent studies within the past few decades have shown that the adult brain contains many
unspecialized progenitor cells that give rise to new neurons (Ming & Song, 2005). These new
neurons are generated in select areas throughout the adult brain, most notably the dentate gyrus
of hippocampal formation. The hippocampus is one of the most important parts of the cognitive
system that plays a crucial role in the maintenance of memories and emotions.

Neurogenic Proliferation
Several studies on laboratory mice have reported that while under baseline conditions,
short sleep deprivation (<1 day) during the normal circadian resting phase did not affect neural
cell proliferation in mice, as seen in Fig. 5 (Van der Borght et al, 2006). This trend seems to
closely follow the same outcome that previous proliferated neurons experienced mentioned
above. However, cell proliferation is significantly suppressed when sleep

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deprivation (or disruption of
sleep) is prolonged and last
for several days or more
(Roman et al, 2005). In order
to further solidify this
hypothesis, a number of
experiments have directly Fig. 5 Effect of sleep deprivation on hippocampal cell proliferation. (B) The
sleep deprivation procedure had no effect on the number of Ki-67 positive cells.
compared the effect of short
(E) BrdU, which had been injected 2 h prior to sacrificing the mice, was
sleep deprivation (<1 day) with incorporated in the same number of cells in sleep-deprived and control mice.
the effects of prolonged sleep restriction ( >3 days) and have confirmed that a reduction in
hippocampal cell proliferation is only found with prolonged sleep restriction or disruption
(Meerlo et al, 2009).

Neurogenic Maturation & Differentiation

Besides the dramatic decline in neurogenic
proliferation, sleep deprivation also has a negative
impact on neurogenic maturation and
differentiation. Several experiments have been
conducted that have showed a decreased amount of
progenitor cells that later express a functional
Fig. 6 Effect of sleep deprivation on phenotype
phenotype that matches already proliferated
expression of surviving cells, with the white columns
pertaining to the well-rested control group and the black neurons. One example can be found in a
columns pertaining to the experimental group. Phenotype experiment that Guzman-Marin conducted. Oddly
of surviving cells were determined by
enough, his data showed that the neurogenic
immunofluorescent triple labeling for BrdU.
maturation to a neuronal phenotype was at its
lowest percentage only after 4 days of complete sleep deprivation, instead of an anticipated 7
days, as seen in Fig. 6 (Guzman-Marin et al, 2007).
Because sleep deprivation has an overall same result on all the steps of neurogenesis,
from neural proliferation to maturation and differentiation, they all equally compromise spatial
memory by decreasing the amount of functional neurons within the brain.

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 Conclusion & Summary

After close analysis of the cumulative experiments, a solid understanding can be achieved
that sleep deprivation (whether in form of limited hours of sleep or fragmentation) leads to an
overall negative effect on neurons, both previously proliferated and those currently going
through one of the many stages of neurogenesis. Neurons present in the hippocampus and cortex
(both of which are closely associated with spatial memory) experienced a range of degrading
histological transformations, ranging from less dense dendritic spines to decreased expressions of
SITR1 and COX. However, after a short period of rest (usually around 3-days), the brain showed
the ability to rejuvenate itself and experience no long-term effects. On top of that, both caffeine
and melatonin doses proved to combat some of the negative effects of sleep deprivation.
Besides directly dealing with neural function, another experiment that proved to provide
another indictor of compromised spatial memory was a decrease in the amplitude of N1, which
can be read by either a PET or fMRI, two modern neuroimaging techniques used today.
Despite all of these discoveries, the general study of neurological transformations that
occur due to sleep deprivation is still a relatively new field of science. At the very least though,
with this more profound understanding on the importance of sleep, perhaps society as a whole
would come to a deeper appreciation for it, thereby leading to a closer adherence to an adequate
daily sleep schedule.

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 Works Cited

Primary Sources:

1) Neuner-Jehle, M., et al. "Characterization and Sleep Deprivation-Induced Expression

Modulation of Dendrin, a Novel Dendritic Protein in Rat Brain Neurons." Journal of
neuroscience research 46.2 (1996): 138-51. Print.

2) Chen, J. -R, et al. "Fatigue Reversibly Reduced Cortical and Hippocampal Dendritic Spines
Concurrent with Compromise of Motor Endurance and Spatial Memory." Neuroscience 161.4
(2009): 1104-13. Print.

3) Chang, Hung-Ming, Un-In Wu, and Chyn-Tair Lan. "Melatonin Preserves Longevity Protein
(Sirtuin 1) Expression in the Hippocampus of Total Sleep-Deprived Rats." Journal of pineal
research 47.3 (2009): 211-20. Print.

4) Cote, K. A., et al. "Waking Quantitative Electroencephalogram and Auditory Event-Related

Potentials Following Experimentally Induced Sleep Fragmentation." Sleep 26.6 (2003): 687-94.
Print.

5) Lorist, M. M., et al. "Influence of Caffeine on Selective Attention in Well-Rested and

Fatigued Subjects." Psychophysiology 31.6 (1994): 525-34. Print.

6) Arrigoni, E., et al. "Adenosine Inhibits Basal Forebrain Cholinergic and Noncholinergic
Neurons in Vitro." Neuroscience 140.2 (2006): 403-13. Print.

7) Van der Borght, K., et al. "Hippocampal Cell Proliferation Across the Day: Increase by
Running Wheel Activity, but no Effect of Sleep and Wakefulness." Behavioural brain research
167.1 (2006): 36-41. Print.

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8) Roman, V., et al. "Sleep Restriction by Forced Activity Reduces Hippocampal Cell
Proliferation." Brain research 1065.1-2 (2005): 53-9. Print.

9) Guzman-Marin, R., et al. "Hippocampal Neurogenesis is Reduced by Sleep Fragmentation in

the Adult Rat." Neuroscience 148.1 (2007): 325-33. Print.

Secondary Sources:

10) Boonstra, T. W., et al. "Effects of Sleep Deprivation on Neural Functioning: An Integrative
Review." Cellular and Molecular Life Sciences 64.7-8 (2007): 934-46. Print.

11) Cabeza, R., and L. Nyberg. "Imaging Cognition II: An Empirical Review of 275 PET and
fMRI Studies." Journal of cognitive neuroscience 12.1 (2000): 1-47. Print.

12) Dunwiddie, T. V., and S. A. Masino. "The Role and Regulation of Adenosine in the Central
Nervous System." Annual Review of Neuroscience 24 (2001): 31-55. Print.

13) Ming, G. L., and H. J. Song. "Adult Neurogenesis in the Mammalian Central Nervous
System." Annual Review of Neuroscience 28 (2005): 223-50. Print.

14) Meerlo, Peter, et al. "New Neurons in the Adult Brain: The Role of Sleep and Consequences
of Sleep Loss." Sleep Medicine Reviews 13.3 (2009): 187-94. Print.

Diagrams:

Fig. 1) Neuner-Jehle, M., et al. "Characterization and Sleep Deprivation-Induced Expression

Modulation of Dendrin, a Novel Dendritic Protein in Rat Brain Neurons." Journal of
neuroscience research 46.2 (1996): 138-51. Print.

10
Fig. 2) Chen, J. -R, et al. "Fatigue Reversibly Reduced Cortical and Hippocampal Dendritic
Spines Concurrent with Compromise of Motor Endurance and Spatial Memory." Neuroscience
161.4 (2009): 1104-13. Print.

Fig. 3) Chang, Hung-Ming, Un-In Wu, and Chyn-Tair Lan. "Melatonin Preserves Longevity
Protein (Sirtuin 1) Expression in the Hippocampus of Total Sleep-Deprived Rats." Journal of
pineal research 47.3 (2009): 211-20. Print.

Fig. 4) Arrigoni, E., et al. "Adenosine Inhibits Basal Forebrain Cholinergic and Noncholinergic
Neurons in Vitro." Neuroscience 140.2 (2006): 403-13. Print.

Fig. 5) Van der Borght, K., et al. "Hippocampal Cell Proliferation Across the Day: Increase by
Running Wheel Activity, but no Effect of Sleep and Wakefulness." Behavioural brain research
167.1 (2006): 36-41. Print.

Fig. 6) Guzman-Marin, R., et al. "Hippocampal Neurogenesis is Reduced by Sleep

Fragmentation in the Adult Rat." Neuroscience 148.1 (2007): 325-33. Print.

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