Restraint stress in biobehavioral research: Recent developments
Tatyana Buynitsky, David I. Mostofsky*
Department of Psychology, Boston University, 64 Cummington Street, Boston, MA 02215, United States
Restraint procedures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Frequency of restraint . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Duration of restraint . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Intensity of restraint . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Sex differences. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Pain in restraint. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Infection, inflammation, and wound healing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Alzheimer’s dementia . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Endocrine, autonomic, and other systems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Prenatal stress . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Acknowledgements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
The literature on stress embraces basic laboratory experimenta-
tion as well as clinical applications in psychology, medicine, and
shown to affect a variety of behaviors including, but not limited to,
physiological (Bao et al., 2008; Harvey et al., 2008; Gorzalka et al.,
2008; Levine, 2005; Ruys et al., 2004; Tabarin et al., 2007),
immunological (Swaab et al., 2005), endocrine (Lemaire et al.,
2000), and developmental processes (Canu et al., 2007), and can
as diabetes (Aguilar-Zavala et al., 2008; Lehman et al., 1991),
depressive-like behaviors (Bauer et al., 2001; Harvey et al., 2005),
tumors (Zhou et al., 1998), neurochemical toxicity in response to
lead exposure (Rossi-George et al., 2008; Yehuda et al., 2008), and
infectious diseases (Glaser et al., 1999b). Recent theory and
experimentation by McEwen and his associates (Conrad et al.,
1996, 1999; Dhabhar et al., 1997; Lehman et al., 1991; Lucas et al.,
2007; Luine et al., 1994; Magarin ˜os and McEwen, 1995a, 1995b;
Wood et al., 2003), Sapolsky et al. (2000) and others have energized
research activities far beyond the modest expectations one might
have predicted from the early influential work of Selye (1976) and
psychosomatic medicine. The impact of stress on aggravating
inflammatory processes has been of particular interest insofar as it
speaks to an understanding of neurological phenomena (e.g. pain)
(Grandin and Deesing, 2002; Schoenfeld, 1981), periodontal
Neuroscience and Biobehavioral Reviews 33 (2009) 1089–1098
A R T I C L EI N F O
Received 21 November 2008
Received in revised form 11 May 2009
Accepted 11 May 2009
A B S T R A C T
In the 15 years since the publication of two previous reviews on restraint stress much advancement has
been made in the field. However, while previous reviews have focused mainly on drug effects, recent
research has focused on broader implications in the health fields. This research has placed an increased
emphasis on stress effects in physiological, immunological, endocrine and developmental processes as
well as theimpact of stress on numerousdisorders. Amajor problem with our review was theinability to
identify a large number of articles focusing on restraint and immobilization, since those keywords were
reviews with extended literature research of this field are required.
? 2009 Elsevier Ltd. All rights reserved.
* Corresponding author. Tel.: +1 617 353 2799; fax: +1 617 353 6933.
E-mail addresses: email@example.com (T. Buynitsky), firstname.lastname@example.org
Contents lists available at ScienceDirect
Neuroscience and Biobehavioral Reviews
journal homepage: www.elsevier.com/locate/neubiorev
0149-7634/$ – see front matter ? 2009 Elsevier Ltd. All rights reserved.
management (Benatti et al., 2003; Gas ˇpers ˇic ˇ et al., 2002; Kesavalu
the influence of stress in the pregnant female affecting irreversible
deficits throughout the lifespan (Canu et al., 2007). Reviews of the
neurobiological, behavioral, and clinical aspects of stress appear in
both journals and published monographs (Ader and Cohen, 1993;
Bao et al., 2008; Fan et al., 1996; Pardon, 2007; Pardon and Rattray,
2008; Swaab et al., 2005).
Experimental models for the induction of both oxidative and
psychological stress exist in a variety of forms that include
procedures incorporating physical as well as psychological
interventions (Bierhaus et al., 2003; Liu and Mori, 1999).
Psychological stress includes an emotional and/or perceptual
response to a stimulus. Such research is particularly important
because stress occurs in the wild, as best exemplified through
social hierarchies in rodents and in primates as described by
Sapolsky’s studies of wild baboons (Sapolsky, 1982). Psychological
stressors may include social order conflicts and competition for
resources as well as restraint and immobilization with accom-
Oxidative stress involves biological damage to tissues. Methods of
physical stress include but are not limited to lack of food or water,
handling and surgical procedures. Physical and psychological
stress can occur together and both can cause outcomes that can be
observed not only behaviorally but also through internal,
biological changes in both animal and human subjects (Bhat
et al., 2007; Glei et al., 2007).
Animal models of stress have proven themselves to be of value
for many reasons. However, as is the case in general programs of
translational research, the utility of these models will often leave
much to be desired. Gaerylewski explained that problems arise
from the ‘‘metabolic, anatomic, and cellular differences’’ between
humans and other animals (2007). Despite the flaws, animal
models offer the best approximation to an understanding of stress
mechanisms in bio-psycho-social organisms. Aside from observa-
tional studies of animals in their natural environment, stress has
been studied by experimentally changing the social order (e.g.
including a dominant aggressive cage mate) (Wood et al., 2003),
introducing a male or femalemember in the litteror cage of a same
sex group, applying electric shock (Grandin et al., 1986), aversive
temperatures (Hashimoto et al., 2001) or loud noise (Khan and
Liberzon, 2004), immersion in water (Grissom et al., 2008),
injectable toxins (Cabib and Castellano, 1997), and surgical
procedures (Sheridan et al., 2004).
Creating conditions of stress in animal experiments continues
to be discussed at conferences and in publications and periodic
reviews in the scientific and clinical literature. Probably the single
most popular experimental preparation for the induction of stress
relies on the use of immobilization or restraint. The extent of such
methodological activities has been summarized in this journal in
1986 (Pare ´ and Glavin, 1986) and again in 1994 (Glavin et al.). In
the 15 years since the last review, ‘‘restraint stress’’ continues to
dominate stress-induced methodology, especially for experiments
that employ rodents as subjects.
research involving stress in wildlife and animal husbandry (Arenz,
1997; Grandinet al., 1986). The outcomesof this research are often
interpreted in the context of the ‘‘fight or flight’’ response, energy
and mineralmetabolismandinteractionswiththe immunesystem
leading to more successful and profitable livestock. Adding pain to
immobilization and restraint stress was observed to produce more
severe changes (Grandin and Deesing, 2002). Grandin et al. (1986)
noted that differing stressors had a variety of implications for the
long-term well being of livestock, and in a field where less aversive
stressors lead to more long-term profits, it was beneficial to select
those that caused the animal less pain or fear.
Furthermore, precise and reliable parametric variations in
employing restraint stress are among the more attractive features
of this procedure, which can take a number of variations, each of
which results in immobilization or restraint. Compared to the
stress induction achieved via drug administration, surgical
interventions, extremes of temperature or acoustic stimulation,
restraint allows for efficiently examining changes in levels of
adrenocorticotropic hormones, corticosterone, fos-protein, desen-
sitization of HPA response and expression of cytokines. A number
of frank behavioral responses such as learning and conditioning
(Jeong et al., 2006; Wu et al., 2007), food intake (Tabarin et al.,
2007), or responses to acoustic startle (Khan and Liberzon, 2004)
and similar environmental challenges can readily be incorporated
along with studies that are formally designed by most life research
Physiological consequences of restraint stress determined from
blood derived assays or direct measurements of cortical micro-
dialysis have been reliably reported in rodents. Extrapolations to
humans, while not usually explicitly suggested, appear reasonable
and representative of stress outcomes found in other species as
well. As noted above, caution must be exercised when attempting
to apply animal study outcomes to human conditions.
1. Restraint procedures
Restraint is a preferred means of stressing animals, largely
because it is straightforward and painless and without lasting
debilitation. Immobilization, a procedural variation of restraint, is
commonly described as restricting range of locomotion but does
not attempt to limit specific limb movement as do most other
restraint techniques. It is important to note that while both
restraint and immobilization constitute physical operations, they
are also important as models for psychiatric stress.
The attraction for using these types of restraint may be due to
the fact that they are inexpensive and rarely if ever involve any
bodily harm to the animal subject, once the period of the restraint
is terminated. This ensures that any long-term effects of stress
observed are due to the stressor which was applied, rather than to
the physical repercussions of an irreversible or chronic injury.
However much psychological stress permeates our society and
however reliable and precise restraint and immobilization
methods may be, these methods may be imperfect models of
the ecological context that is usually experienced.
A number of variations in effecting restraint have been
published. No comparative studies of the relative merits favoring
any one procedure have been reported. Inter-study comparisons
are often difficult if not impossible to determine, e.g. when some
experimenters choose to subject their animals for a single session
of 5 min (Jackson and Moghaddam, 2006; Kang et al., 2007), and
others use sessions lasting up to three weeks (Davydov and Shvets,
2003; McLaughlin et al., 2007). The psychological and physiolo-
gical changes associated with restraint appear to result from the
distress and aversive nature of having to remain immobile, rather
than any concurrent activation of pain mechanisms or irreversible
discomfort (Grandin and Deesing, 2002). Common to all methods
of restraint is the restriction and immobilization of movement and
include (but are not necessarily limited to) the following: (a) the
entire animal is placed in a rigid Plexiglas cone; (b) the entire
animal is placed in a Decapicone (a tapered, conical, plastic film
tube); (c) the limbs are taped to a board or tied to pads; (d) the
entire animal is placed in a wire mesh restrainer; (e) a plastic or
Plexiglas panel is used to restrict motor movements to a corner of a
cage; (f) the animal is placed in a plastic bottle or a restraint cage;
(g) the entire animal is wrapped with wire mesh gauze; (h) the
animal is rolled in a cloth towel, with only the snout and tail
showing. Some reports refer to the use of an ‘‘immobilization’’ bag
T. Buynitsky, D.I. Mostofsky/Neuroscience and Biobehavioral Reviews 33 (2009) 1089–1098
which would appear to be more commonly described as a restraint
holder; however, a detailed description of its construction was not
provided (Vyas et al., 2002) (Table 1).
In certain instances, restraint and immobilization will not
continue to increase monotonically with the continued application
of stressors. A peak may be reached and the animal may adapt to
stress, causing it to stop or attenuate a SR. This may be due to
restraint and immobilization can be additionally assessed by
incorporating behavioral measures along with biophysiological
ones. Since lack of a SR may be due to habituation (Dallman, 2007),
the frequency of exposures is significant and interrupting the chain
of repeated restraint sessions may serve to minimize the likelihood
of habituation. Dallman et al. (2005) explained, ‘‘The hypothalamo–
pituitary–adrenal (HPA) axis is a prototype of neuroendocrine
systems with inhibitory feedback loops mediated by the hormone
secreted from a remote target gland.’’ This raises the question, to
what extent do physiological mechanisms follow the rules of
learning in studies of learned helplessness (Seligman, 1975).
As seen in Table 1, the most common physiological changes
observed include increases in ACTH and corticosterone levels,
however routine behavior effects are just as prevalent. Very often
these involve decreases in memory acquisition and retention as
well as changes in motivation. Gameiro et al. (2006) reported such
changes in ACTH and corticosterone along with hyperalgesia
which was eased with fluoxetine. While this conclusion is
important in terms of observing changes in serotenergic and
opioid systems, it also has important implications for pharma-
ceutical development. Motivation involves both internal desires as
well as the subject’s responses to competition from peers.
Decreased levels of motivation can be inferred by the rodent’s
an animal at the top of the social hierarchy is no longer able to act
aggressively and dominantly. A wire mesh restrainer has been a
particularly popular in showing a decrease in attacks and also a
lowered frequency of wounds in comparison to the social situation
that existed before the administration of the stressor. Wood et al.
(2003) suggested that such changes have implications for the
incidence of aggression and can be used to analyze stress-
responsive disorders. Based on the stress implications that have
been determined, we could expect SR to affect all organ systems.
Although many early studies have not emphasized the direct
effects of stress on the brain, recent research has been concerned
with the immune and endocrine systems which has generated
speculation concerning specific human illnesses that may exist.
The use of a restraint cage has shown increased levels of anorectic
responses with the increased latency to eat and decreased meal
size (Wang et al., 2002). Further, in animal subjects who have
regularly ingested ethanol, the levels of intake have increased after
each consecutive application of stress. However, in addition to
anorexia and alcoholism, the major deficit that has been addressed
is a decreased immune response as found in a 2002 study by Wang
et al. where the m-opioid receptor is involved in restraint stress-
induced immune changes. They concluded that research on the
mechanism that controls stress in the immune system could have
implications in therapeutic methods by allowing the blocking of
the m-opioid receptor.
Stress effects based on type of restraint/immobilization.
(a) Rigid Plexiglas tube or
# Motor activity, motivation, acquisition, memory retention and
consolidation and reactivity, " c-fos-positive neurons and nNOS/c-fos
double labeled neurons, " levels of thiobarbituric acid in brain and
retina, " ACTH and corticosterone secretion, " in fos-protein, c-fos
mRNA, and zif268 mRNA, " nitrite levels, # licking/guarding reflexes,
" escape responses, " TNF-a levels and TNF-a converting enzyme
activity, induced immune alterations, changes in CMI function
Abidin et al. (2004), Akana et al. (1992), Bauer et al. (2001),
Bizon et al. (2001), Bowman et al. (2006), Cabib and
Castellano (1997), Canu et al. (2007), Cecchi et al. (2002),
Chadda and Devaud (2005), Chou-Green et al. (2003),
Colomina et al. (1999), Derin et al. (2006), Dhabhar et al. (1997),
Engler et al. (2005), Freeman et al. (2007), Gameiro et al. (2006),
Gas ˇpers ˇic ˇ et al. (2002), Grissom et al. (2008), Head et al. (2006),
Helm et al. (2004), Hsu et al. (1998), Hunzeker et al. (2004),
King et al. (2007), King et al. (2003), Kitraki et al. (2004),
Lee et al. (2009), Lemaire et al. (2000, 2006), Luine et al. (1994),
Madrigal et al. (2003), Munhoz et al. (2004), Pace et al. (2005),
Pecoraro et al. (2004), Pedersen et al. (1999), Rissman et al.
(2007), Rojas et al. (2002), Rossi-George et al. (2008),
Sarkar et al. (2007),
Sheridan et al. (2004), Wang et al. (2002),
Weinberg et al. (2007), Zhou et al. (1998)
Belda et al. (2004), Dal-Zotto et al. (2004), Davydov and Shvets
(2003), Gagliano et al. (2008), Kumari et al. (2007), Liu et al.
(1996), Liu et al. (1994), S ahin and Gu ¨mu ¨s ¸lu ¨ (2007), Yaras et al.
(2003), Yoshihara and Yawaka (2008), Zorzet et al. (1998)
(c) Taped limbs to surface
" ACTH response, sensitization of HPA axis but desensitization after a
previous exposure, " corticosterone levels, " lipid peroxidation,
# efficiency in the heart, # immune function, # endogenous antioxidant
defenses, long-term effects on growth of developing rats and stress
response of the HPA axis
# Spatial memory, # body weight gain, CA3 dendritic retraction,
# levels of motivation, " hippocampal-dependent behavior,
# open field exploration, # aggressive behavior and wounding
(d) Wire mesh restrainer
Bhat et al. (2007), Conrad et al. (1999), Conrad et al. (1996),
Galea et al. (1997), Inoue et al. (1993), Kleen et al. (2006),
Magarin ˜os and McEwen (1995a), Magarin ˜os and McEwen
(1995b), McDougal et al. (1983), McLaughlin et al. (2007),
Miyahara et al. (1998), Wood et al. (2003),
Zafir and Banu (2008)
Jackson and Moghaddam (2006)
Ada ´mekova ´ et al. (2003), Ceballos et al. (2006), Costa et al.
(2005), Fujioka et al. (1999), Harvey et al. (2005),
Hashimoto et al. (2001), Jeong et al. (2006), Kang et al. (2007),
Lehman et al. (1991), Lynch et al. (1999), Nakajima et al. (2006),
Popovic ´ et al. (1996), Ruys et al. (2004), Tabarin et al. (2007)
Benatti et al. (2003), Fan et al. (1996), Grandin et al. (1986),
Khan and Liberzon (2004), Lucas et al. (2007), Taylor et al.
(1995), Vyas et al. (2002)
(e) Clear panel
(f) Restraint cage or box
" Firing rate of neurons initially but then return to normal
# Levels of leptin, " levels of corticosterone, " hippocampal nitrogen
oxides, " ethanol intake, anorectic response, analgesic state,
" pain response, " CRF
(g, h) Unspecified/other
" Acoustic startle response, immune suppressive protein generated,
allostatic attenuation of the mesolimbic dopaminergic system,
" flinching, " arterial pressure and heart rate, abnormalities in blood
pressure systems, reluctant acceptance of reward, decreased fear
T. Buynitsky, D.I. Mostofsky/Neuroscience and Biobehavioral Reviews 33 (2009) 1089–1098
2. Frequency of restraint
As mentioned in previous reviews, the frequency of a stressor
(defined as the number of sessions per day or week) has been a
focal point of many studies with little concern for the minimum
required to produce the SR. In addition,intheir review, Glavin et al.
(1994) reported that stress occurring from restraint is not
necessarily a linear phenomenon of frequency but rather that
the severity of a method of restraint determines whether or not the
subject had the opportunity to habituate to the stressor and to
decrease the strength of the SR. Other studies have been conducted
with identical stressors with variations in both duration and
frequency. Such studies have shown that certain biological
responses appear to increase with a consistent application of
stress while others will decrease. For example, Munhoz et al.
(2004) placed rodents in a conical Decapicone restrainer for 6 h a
day for either 7 or 14 consecutive days. Their results showed that
enzymatic activity returned to basal values a day after 7 days of
stress but did not have the same effect after 14 days. Additionally,
increased malondialdehyde (a marker of oxidative stress) accu-
mulationin cortex returnedto normal a dayafter stress, onlyin the
seven-day stress group. TNF-a (tumor necrosis factor) accom-
panies inflammation and it might be expected that any amount of
stress would always bring about this change and increase
consistently. However, TNF-a levels increased mainly after the
14-day group even though there was an increase in the TNF-a
converting enzyme activity in both groups. This study showed that
not all exposures to stress were enough to elicit the expected
response and that different outcomes appear in enzyme activity
regardless of the level of increase in the TNF-a levels themselves.
As seen in Table 2, stress frequency affects the outcome
variables as well as the range of different findings. Single session
applications of stress seem to be more common and are often
followed by sacrifice of the animal. However, a limited number of
the studies are concerned with the long-term effects of stress after
a single application. Often a three week, daily stress period is
utilized and reported findings generally tend to have been taken at
the conclusion of the stress period rather than during regular
Stress effects based on frequency of restraint/immobilization.
" ACTH response and corticosterone secretion, # activity and exploration,
" c-fos-positive neurons and nNOS/c-fos double labeled neurons,
# ENK-mRNA levels but " in DAT and D2r binding in the dorsal striatum,
desensitization of the HPA response, no long-term effects on growth of the
HPA axis, # lick/guard reflexes but " escape responses, " pain response,
" acoustic startle response, " firing rate of hippocampal nitrogen oxides,
# efficiency in induction of free radial processes in the heart, " anxiety,
" stroke volume, # heart rate, # memory retention and consolidation,
" lipid peroxidation, " protein oxidation, abnormalities in blood pressure
control system, " stomach bleeding, # plasma iron-binding and
iron-oxidizing, " cortisol and non-esterified fatty acids, corticotropin
releasing factor-like immunoreactivity, # stress with experience,
changes in CMI structure
" Firing rate of neurons followed by #, # food intake followed by ",
" plasma corticosterone and pituitary proopiomelanocortin
Immediate anorectic response, " levels of ACTH and corticosterone,
" anxiety behavior, " serum adenosine deaminase levels
Arborgast et al. (1994), Arenz (1997), Bauer et al. (2001), Belda et al. (2004),
Bizon et al. (2001), Bosch et al. (2000), Cabib and Castellano (1997), Cecchi
et al. (2002), Chappell et al. (1986), Costa et al. (2005), Davydov and Shvets
(2003), Dhabhar et al. (1997), Echeverry et al. (2004), Gagliano et al. (2008),
Grandin et al. (1986), Haas and George (1988), Harvey et al. (2005),
Hsu et al. (1998), Hunzeker et al. (2004), Inoue et al. (1993),
Kang et al. (2007), Khan and Liberzon (2004), King et al. (2003),
King et al. (2007), Kumari et al. (2007), Liu et al. (1994, 1996),
Lucas et al. (2007), Madrigal et al. (2003), McDougal et al. (1983),
Miyahara et al. (1998), Pace et al. (2005), Popovic ´ et al. (1996),
Rossi-George et al. (2008), Sarkar et al. (2007), Seo et al. (2006),
Strausbaugh et al. (1999), Taylor et al. (1995), van Rensburg et al. (2006),
Weinberg et al. (2007), Yoshihara and Yawaka (2008)
Akana et al. (1992), Dal-Zotto et al. (2004), Jackson and Moghaddam
(2006), Wang et al. (2002), Wu et al. (2007), Zhou et al. (1998)
3Fujioka et al. (1999), Gameiro et al. (2006), Hashimoto et al. (2001),
Laconi et al. (2000), Miyahara et al. (1998), Rojas et al. (2002),
Tabarin et al. (2007)
Chou-Green et al. (2003), Freeman et al. (2007), Masuda et al. (2004)
# In food intake in older mice, # of the TG-resident HSV-specific CD8+T
cells and a functional compromise of those cells that survive
" Responses to pain behavior, " levels of ACTH and corticosterone,
" anxiety behavior, " nociceptive responses/hyperalgesia, " density of
neurons expressing NADPH-d and nNOS in the amygdaloid nuclei,
" ethanol intake, " struggling that decreased over time
# Growth of the HPA axis during development in the postnatal period
# Behavioral agitation, # cortisol secretion, " sensitivity to
dexamethasone, enzymatic activity returned to basal levels after
" Struggling that decreased over time, # wound closure,
# pro-inflammatory gene expression
# Spatial memory, allostatic attenuation of dopaminergic systems,
" atherogenic plaque progression, immunological deficits, " CRF
Short term protection against seizure induction
" TNF-a levels, " TNF-a converting enzyme activity, " atherogenic
plaque progression, habituated endocrine and immune responses,
# motor resistance and coordination
" Plasma corticosterone levels, " reactive substances in all tissues,
" lipid peroxidation, " GSH-Px activity in the brain
" Plaque pathogenesis
CA3 apical dendritic retraction, # spatial memory, # body weight gain,
" thiobarbituric acid reactive substance levels in brain and retina, latencies
of visual evoked potentials provoked, # leptin levels, " corticosterone levels,
# reward related motivation, " nitrite levels, # memory acquisition and spatial
memory, # aggressive behavior, # wounding frequency when aggressive
behavior occurred, # open-field exploration, " hippocampal-dependent and
hippocampal independent behavior, " gum inflammation
Atrophied thymus and spleen
" Atherogenic plaque progression
" Degradation in periodontal tissues
" Bone loss when combined with an injection of nicotine
Echeverry et al. (2004), Gameiro et al. (2006), Grissom et al. (2008),
Helm et al. (2004), Lynch et al. (1999), Pecoraro et al. (2004),
Seo et al. (2006)
Yoshihara et al. (2007)
Ada ´mekova ´ et al. (2003), Engler et al. (2005),
Munhoz et al. (2004), Ruys et al. (2004)
8Grisson et al. (2008), Head et al. (2006), Sheridan et al. (2004)
10 Dhabhar et al. (1997), Inoue et al. (1993), Li et al. (2002),
Lucas et al. (2007), McLaughlin et al. (2007), Vyas et al. (2002)
Chadda and Devaud (2005)
Bauer et al. (2001), Bhat et al. (2007), Colomina et al. (1999),
Lehman et al. (1991), Li et al. (2002), Magarin ˜os and McEwen (1995a),
Munhoz et al. (2004), Rissman et al. (2007)
S ahin and Gu ¨mu ¨s ¸lu ¨ (2007), Yaras et al. (2003)
Lee et al. (2009)
Abidin et al. (2004), Bowman et al. (2006), Campan et al. (1997),
Ceballos et al. (2006), Conrad et al. (1996), Conrad et al. (1999),
Derin et al. (2006), Galea et al. (1997), Kleen et al. (2006),
Kitraki et al. (2004), Luine et al. (1994), Magarin ˜os and McEwen
(1995a, 1995b), Magarin ˜os and McEwen (1995a), McLaughlin
et al. (2007), Wood et al. (2003), Zafir and Banu (2008)
Nakajima et al. (2006)
Li et al. (2002)
Gas ˇpers ˇic ˇ et al. (2002)
Benatti et al. (2003)
T. Buynitsky, D.I. Mostofsky/Neuroscience and Biobehavioral Reviews 33 (2009) 1089–1098
intervals. Evidence concerning long-term effects of single applica-
tion restraint stress have not systematically been examined and
therefore remain inconclusive.
3. Duration of restraint
The duration of restraint refers to the length of time of exposure
during each session. It is important to note that the increase in the
duration of restraint does not necessarily imply an increase in the
stress experience. At times, a researcher interested in social
interactions will decide that 5 min of restraint is sufficient and
then observe interactions of the rodent with other subjects. At
other times, restraint may continue for 6 h at which time the
rodents brain is observed without an opportunity to return to its
normal metabolic state (see Table 3).
4. Intensity of restraint
Intensity is the extent of immobilization or restraint that the
rodent is subjected to (e.g. limited mobility in a wire mesh
restrainer where the animal can turn himself around vs. complete
absence of mobility in a Plexiglas tube). Variation in the types of
restraint intensity employed can suggest a change in the level of
psychological stress the rodent experiences. Another difference in
intensity can be observed when comparing stressors that are
physical in nature, with those that are caused by chemical means
(such as causing temporary paralysis). Arenz’ (1997) study of fox
squirrels is one of the few where the different types of restraint
have been addressed at all. Arenz used two stressors, a simple
physical restraint (which is not specified) and a ketamine-
hydrochloride injection (a strong anesthetic that led to immobi-
lization but not causing a total body paralysis). When comparing
the two types of restraint, Arenz noted that after the injection, but
before recovery, the animals became increasingly more active and
difficult to handle whereas post physical restraint, they were
relatively easy to control. It could not be concluded that this is
because physical restraint causes an animal to feel a heightened
sense of danger, causing the need to escape, or if it is because with
physical restraint, the animal is able to discriminate what is
preventing movement and can then attempt to counter it.
Researchers have speculated that the animals that are physically
restrained seem to cope better because they are able to learn the
cause of the restraint, unlike those that are simply given an
injection making them unable to move.
Grandin et al. (1986) examined two forms of restraint in ewes,
i.e. electro-immobilization and a mechanical restraint table. The
ewes that were restrained physically became accustomed to the
restraint and ceased to fight being placed on the table. However,
those that were restrained both mechanically and via electro-
immobilization displayed more fear of the table and accepted food
as a reward more hesitantly (and sometimes not at all) after the
table restraint was experienced. This confirms Arenz’ notion that
Stress effects based on duration of restraint/immobilization.
Under 15 min Significantly # licking/guarding reflexes but " escape responses, " in firing
rate of neurons, " ACTH and corticosterone secretion, " fos-protein,
c-fos mRNA, and zif268 mRNA expression, # food intake, " CRF content,
hesitancy to accept feed reward
" Number of c-fos-positive neurons and nNOS/c-fos double-labeled neurons,
" corticosterone, # leptin, " ACTH and corticosterone secretion, # food intake,
# licking/guarding responses, " learned escape responses, " operant sensitivity,
" ethanol intake, " difficulty in handling, # memory retention and consolidation,
" expression of the gene for Type I diabetes, " development of diabetes
Cecchi et al. (2002), Dal-Zotto et al. (2004), Fujioka et al. (1999),
Grandin et al. (1986), Haas and George (1988), Jackson and
Moghaddam (2006), King et al. (2007), Pace et al. (2005)
Arenz et al. (1997), Cabib and Castellano (1997), Ceballos et al.
(2006), Chou-Green et al. (2003), Dal-Zotto et al. (2004),
Gameiro et al. (2006), Hashimoto et al. (2001), Inoue et al. (1993),
King et al. (2003), Lehman et al. (1991), Lynch et al. (1999),
Sarkar et al. (2007), van Rensburg et al. (2006),
Weinberg et al. (2007), Zhou et al. (1998)
Akana et al. (1992), Bauer et al. (2001), Belda et al. (2004),
Bosch et al. (2000), Cabib and Castellano (1997), Chadda and
Devaud (2005), Davydov & Shvets (2003), Gameiro et al. (2006),
Grissom et al. (2008), Helm et al. (2004), Pace et al. (2005),
Rissman et al. (2007), Strausbaugh et al. (1999)
Canu et al. (2007), Derin et al. (2006), Lemaire et al. (2000),
Lemaire et al. (2006), Rossi-George et al. (2008),
Wu et al. (2007), Yoshihara and Yawaka (2008)
Abidin et al. (2004), Bizon et al. (2001), Cabib and Castellano
(1997), Colomina et al. (1999), Costa et al. (2005),
Gameiro et al. (2006), Hsu et al. (1998), Inoue et al. (1993),
Masuda et al. (2004), Pace et al. (2005), Seo et al. (2006),
Tabarin et al. (2007), Vyas et al. (2002), Weinberg et al.
(2007), Zorzet et al. (1998)
Ada ´mekova ´ et al. (2003), Benatti et al. (2003),
Chappell et al. (1986), Dhabhar et al. (1997), Echeverry et al. (2004),
Fujioka et al. (1999), Gagliano et al. (2008), Harvey et al. (2005),
Hashimoto et al. (2001), Hunzeker et al. (2004), Inoue et al. (1993),
Kang et al. (2007), Khan and Liberzon (2004), Lee et al. (2009),
Lucas et al. (2007), McLaughlin et al. (2007),
Miyahara et al. (1998), Pecoraro et al. (2004), Ruys et al. (2004),
Weinberg et al. (2007), Yaras et al. (2003), Zafir and Banu (2008)
Arborgast et al. (1994), Bhat et al. (2007), Bowman et al. (2006),
Conrad et al. (1999), Conrad et al. (1996), Engler et al. (2005),
Freeman et al. (2007), Galea et al. (1997), Gas ˇpers ˇic ˇ et al. (2002),
Head et al. (2006), Jeong et al. (2006), Kitraki et al. (2004),
Kleen et al. (2006), Kumari et al. (2007), Liu et al. (1996),
Liu et al. (1994), Luine et al. (1994), Madrigal et al. (2003),
Magarin ˜os and McEwen (1995a, 1995b), McLaughlin et al. (2007),
Munhoz et al. (2004), Nakajima et al. (2006), Rojas et al. (2002),
Sheridan et al. (2004), Taylor et al. (1995), Wang et al. (2002),
Wood et al. (2003)
" ACTH and corticosterone, " in hyperalgesia, " fos-protein, c-fos mRNA,
and zif268 mRNA expression, desensitization of the HPA response,
# efficiency in the induction of free radial processes in the heart,
habituated endocrine and immune responses, " anxiety, " stroke volume,
# heart rate, # memory retention and consolidation
# Levels of motor activity, long-term effects on growth of developing rats,
" levels of thiobarbituric acid reactive substances in brain and retina
1 h to 1 h and
Anorectic response with " latency to eat and # meal size, " ACTH levels,
" corticosterone levels, " anxiety behavior, acute stress " analgesic state,
" fos-protein, c-fos mRNA, and zif268 mRNA expression, " nitrite levels in
frontal cortex and hippocampus, attenuation of immune functions, # motor
resistance and coordination, # memory retention and consolidation
2 h to 5 h and
" ACTH response, # levels of activity and exploration, # ENK-mRNA levels,
" DAT and D2r binding, CA3 apical dendritic retraction, # spatial memory,
# body weight gain, restraint induced fra-2 expression, " hippocampal nitrogen
oxides, " density of neurons expression NADPH-d and nNOS, " acoustic startle
response, # behavioral agitation and cortisol concentrations, " sensitivity to
dexamethasone, " plasma corticosterone concentrations, " serum adenosine
deaminsane levels, " corticosterone levels, " CRF, corticosterone releasing
factor-like immunoreactivity detected
CA3 apical dendritic retraction, # spatial memory, # body weight gain, # reward
related motivation, " malondialdehyde accumulation, " TNF-a levels and TNF-a
converting enzyme activity, # stress-induced malondialdehyde accumulation,
# aggressive behavior, # wounding frequency when aggressive behaviors occur,
restraint induced immune alterations, " freezing to context and tone, # open-field
exploration, # spatial memory performance on the Y maze, " lipid peroxidation,
abnormalities in blood pressure control systems, " cortisol and non-esterified
fatty acids, " stomach bleeding, # plasma iron-binding and iron-oxidizing
T. Buynitsky, D.I. Mostofsky/Neuroscience and Biobehavioral Reviews 33 (2009) 1089–1098
physical restraint is less traumatic in the long term compared to
restraint techniques that do not allow the subject to discriminate
the source of immobilization. However, since the Grandin et al.
study does not implement a control, these conclusions can be
challenged on the grounds that the effect of the shock of electro-
immobilization apart from any other kind of immobilization can
lead to these behavioral consequences.
5. Sex differences
Although not always intuitively obvious, some researchers
found that sex differences in the physiological reactions that
rodents exhibit when stressed will occasionally vary (Bosch et al.,
2000; Masuda et al., 2004; Chadda and Devaud, 2005). The results
of these studies indicated that when stressed, males showed
habituation while females showed sensitization.
More specific endocrine differences include higher basal and
stress levels in the plasma corticosterone and corticosteroid-
binding globulin in adult females as well as a longer period of
elevation in plasma corticosterone levels in the SR of male rats
(Galea et al., 1997). Additionally, with the implementation of
restraint stress, glucocorticoid receptors decreased in the hippo-
campus of male rats while they increased in those of female rats
(Kitraki et al., 2004).
6. Pain in restraint
Stress and pain have a complex relationship where pain can
cause stress, stress can increase pain, or the two can be observed
concurrently such as in hyperalgesia. A fundamental problem with
studying pain in animals is that it is undetermined whether pain is
a response, a drive, or a stimulus (Schoenfeld, 1981). The
distinction between animals’ experience of pain and suffering or
whether this experience is related to the size and capacity of the
brain remains to be determined. Grandin and Deesing describe
some studies that assert that smaller brains do not feel as much
pain as intensely as more complex organisms (2002). From this
perspective the experience of pain could be speculated to result
from the quantity of connections between sub-cortical structures
and the nervous system, thus suggesting that small mammals are
less responsive to pain. However, Grandin and Deesing argue that
this is not the case and that complex stress experiences are
particularly prevalent in rodents. They cite that the removal of a
variety of pain areas in the brain, particularly in the prefrontal
cortex, will cause the animal to no longer experience pain but will
still condition them to a fear or SR. Even when no links exist
between the nervous system and pain receptors, the animals still
experience stress, thus showing that even in the smallest animals
where pain is not a likely variable, suffering can exist from the
experience of conditioned stress.
King and Devine (2007) and King et al. (2003) cite the
suppression of lick-guard reflexes which suggests hyporeflexia
but simultaneously shows hyperalgesia through escape responses.
In their studies they incorporated restraint and the application of
restraint, the rat has less immediate response to the pain it is
experiencing but is more likely to attempt to ultimately flee the
situation. The findings of Gameiro et al. (2006) support this theory,
and conclude that hyperalgesia and the responses observed to
morphine, naloxone, and fluoxetine stem from the serotenergic
and opioid systems.
Taylor et al. (1995) examined changes in systems that control
blood pressure and observed responses such as flinching, arterial
pressure, and heart rate levels that suggest that changes in these
variables cause a greater response to pain when the stimuli are
repeated and consistent. Thus, little variation exists in the
They concludedthat post-
expectations of pain response as it occurs after a period of
immobilization where helplessness is likely learned.
7. Infection, inflammation, and wound healing
Stress, including restraint stress, affects the immune system
including the reactions of cytokines, celladhesion molecules, and T
cell responses when the body is fighting viral and bacterial
infections (Wheway et al., 2007). The neuroendocrine responses to
infections such as influenza, periodontitis, and herpes vary;
nonetheless, they all show that stress consistently decreases the
healing time and the body’s ability to fight infection.
Studies in periodontitis have shown that restraint stress by
itself does not result in periodontal disease but instead lengthens
the amount of time to decrease inflammation. One such study
showed that periodontal tissues break down faster when rats were
exposed to the combination of both restraint and surgical stress or
restraint and bacterial infection (Houri-Haddad et al., 2003). When
stress was caused by both restraint and ligation there was higher
effects were not seen when both stressors were not implemented
together. In a similar study by Benatti et al. (2003) nicotine was
found to have enhanced effects on periodontitis when immobiliza-
tion stress was implemented. With nicotine expected to increase
the development of inflammation, their study reported the
greatest bone loss in rats was found when stressed (immobiliza-
tion) and nicotine injections were used to accelerate periodontitis.
In the case of influenza, restraint stress negatively affected the
expression of cytokines, cell adhesion molecules, and cell surface
receptors. Restraint stress was found to slow the rate of cell
migration to the location of the influenza infection (Echeverry
et al., 2004). Hunzeker et al. (2004) showed a decrease in natural
killer cell function recruited and activated by cytokines and an
increase in viral replication after restraint stress. Studies of herpes
found similar effects (Freeman et al., 2007) including a decrease of
about 65% in the ability of cells to produce the antiviral cytokine
IFN-g that responds to the virus.
Finally, in the case of bacterial infections, restraint stress was
to slow the closure of wound healing by a factor of 2 (from 3 to 7
days) (Head et al., 2006) and to increase opportunistic infections
(Rossi-George et al., 2008). When comparing restraint stress to
restraint stress was found to slow wound healing while the social
stressor had the same effects as the control (Sheridan et al., 2004).
Similar stress effects on infection and inflammation responses in
humans have been studied in models not utilizing restraint
(Christian et al., 2002; Glaser et al., 1999a; Kiecolt-Glaser et al.,
1995; Kiecolt-Glaser and Glaser, 2001; McGuire et al., 2006).
8. Alzheimer’s dementia
The role of stress in the genesis and evolution of Alzheimer’s
dementia is also an issue that has been of great interest to society.
A causal role of stress in the onset and progression of Alzheimer’s
disease pathology has been suggested (Sotiropoulos et al., 2008).
A study by Jeong et al. (2006) found that cognitive impairments
developed faster as a result of chronic immobilization stress.
Stressed animals had more and denser vascular and extracellular
deposits containing amyloid beta peptide and carboxyl-terminal
fragments, elevated levels of neurodegeneration and tau phos-
phorylation, and increased intraneuronal Ab and APP-CTFs
immunoreactivities, all of which are indicators of Alzheimer’s
disease. Additionally, van Rensburg et al. (2006) found a higher
oxidation status in subjects with Alzheimer’s exposed to restraint
T. Buynitsky, D.I. Mostofsky/Neuroscience and Biobehavioral Reviews 33 (2009) 1089–1098
Other studies have shown additional debilitating effects of
restraint stress on Alzheimer’s disease including oxidative damage
(Kumari et al., 2007), down regulation of MMP-2 (an enzyme that
decreases Ab levels) expression in the brain (Lee et al., 2009), and
elevation of plasma glucocorticoid levels and hypoglycemia
(Pedersen et al., 1999). However, conclusive evidence of a
stress-Alzheimer’s connection has been seriously questioned
(Swaab et al., 2005; Popovic ´ et al., 1996).
9. Endocrine, autonomic, and other systems
may also preferentially affect certain organ systems. Furay et al.
(2009) found that the forebrain glucocorticoid receptor is partially
responsible for the ‘‘negative feedback regulation of HPA axis
responses to psychogenic stressors’’ and also that ‘‘chronic stress-
induced alterations in the HPA axis function are mediated by
mechanisms independent of the forebrain glucocorticoid recep-
tor.’’ They concluded that the forebrain glucocorticoid receptor is
largely responsible for neuroendocrine function.
Glucocorticoids were also found to have a role in habituation.
Dallman (2007) noted that compared to rats experiencing a
stimulus for the first time, those exposed to a stimulus multiple
times have decreased activity in the hypothalamic paraventricular
nucleus, a region involved in the release of corticotrophin-
releasing factor (CRF). Additionally, corticotropin-releasing factor,
vasopressin and thus ACTH and corticosterone release are
Neuroendocrine responses are also known to have a connection
to obesity and the intake of comfort foods. The effects of stress and
glucocorticoids suggest that restraint increases the desire for
comfort foods which then decreases the activity in the HPA axis
(Pecoraro et al., 2004). Comfort foods (foods high in sugars and/or
fats) have been found to increase obesity and corticotropin-
releasing factor in the hypothalamus of rats and as a result,
Dallman et al. (2003) speculate that people eat comfort foods to
‘‘reduce the activity in the chronic stress-response network with
its attendant anxiety’’ and suggest implications for anorexia
nervosa and Cushing syndrome (Dallman et al., 2004).
10. Prenatal stress
Prenatal stress, especially in humans, is thought to cross the
placenta and affect the fetus, possibly causing premature birth,
slowed brain development, and a higher risk of mental illness in
the progeny of a highly stressed mother. This is a reasonable
speculation considering that the exposure to both prenatal lead in
the presence of prenatal stress is more likely to have detrimental
results as compared to exposure to lead alone (Rossi-George et al.,
2008). Studies by Fujioka et al. (1999) and Hashimoto et al. (2001)
showed that restraint stress causes behavioral changes in the
offspring of stressed mothers. Fukioka et al. found that maternal
stress can change the organization of fetal brain neurons and that
long-term stress was most likely to be destructiveto corticotropin-
releasing-factor neurons in the paraventricular nucleus. It is also
important to note that prenatal restraint stress had similarly
negative effects on the corticosterone response as did chemical
injection of lipopolysaccharide suggesting that a multitude of
stressors can cause negative effects in offspring, as shown by
Hashimoto et al.
Several studies have also shown that prenatal restraint stress
affects cell proliferation, particularly in the hippocampus and in
of prenatal stress include ‘‘the survival rate of newborn cells, the
and memory (Wu et al., 2007), diminished feedback mechanisms of
the HPA axis (Maccari and Morley-Fletcher, 2007), disruption of
circadian rhythms, and altered neuroplasticity (Jackson and
Moghaddam, 2006). In general, prenatal stress was shown to have
long-term consequences in the offspring of the stressed mothers.
However, it would seem that the examination of long-term
consequences of stress in study designs that utilize stressing the
expecting mother and then study the long-term effects on the
offspring cannot be used to compare the long-term effects at two
different time points for the same animal.
Methodological features of the various studies often cannot be
compared because of the design that may be unique to the study.
Additionally, previous reviews, while unquestionably thorough,
were also unable to include all existing research for the period that
they undertook to survey. While this present review includes
further elaboration on previous restraint and immobilization
research, it is most important to note that further research will be
necessary to address emerging issues such as anti-stress, as for
example in the Bhat et al. (2007) study of pyramidal energy
(energy generated when a subject is contained inside a pyramid-
shaped box which is believed to have an anti-stress effect) on
cortisol levels, oxidative stress, and antioxidants. There also
remains much to be learned about the effects of restraint stress
on previous exposure to carcinogens where stress has been found
to speed up the development of carcinomas (Ada ´mekova ´ et al.,
2003; Laconi et al., 2000). An additional recent study has
demonstrated the causal role of glucocorticoids in oxidative
processes and has been the first to connect it to the performance of
vital organs such as the brain, liver, and heart (Zafir and Banu,
2008) with further research being necessary.
However, searches of the restraint literature are seriously
limited by the absence of the term ‘‘restraint’’ or ‘‘immobilization’’
in either title, abstract, or keywords of many published reports, e.g.
Chappell et al. (1986) and Helm et al. (2004).
We thank Mary Foppiani, Librarian at the Boston University
Science and Engineering Library.
Abidin, I., Agar, A., Gu ¨mu ¨slu ¨, S., Aydin, S., O¨ztu ¨rk, O., Sahin, E., 2004. The effects of
chronic restraint stress on spatial learning and memory: relation to oxidant
stress. International Journal of Neuroscience 114, 683–699.
Ada ´mekova ´, E., Markova ´, M., Kubatka, P., Bojkova ´, B., Ahlers, I., Ahlersova ´, E., 2003.
NMU-induced mammary carcinogenesis in female rats is influenced by
repeated psychoemotional stress. Neoplasma 50, 428–432.
Ader, R., Cohen, N., 1993. Psychoneuroimmunology: conditioning and stress.
Annual Reviews of Psychology 44, 53–85.
Aguilar-Zavala, H., Garay-Sevilla, M.E., Malacara, J.M., Pe ´rez-Luque, E.L., 2008.
Stress, inflammatory markers and factors associated in patients with type 2
diabetes mellitus. Stress and Health 24, 49–54.
Akana, S.F., Dallman, M.F., Bradbury, M.J., Scriber, K.A., Strack, A.M., Walker, C.D.,
1992. Feedback and facilitation in the adrenocortical system: unmasking
facilitation by partial inhibition of the glucocorticoid response to prior stress.
Endocrinology 131, 57–68.
Arborgast, B.W., Neumann, J.K., Arborgast, L.Y., Leeper, S.C., Kostrzewa, R.M., 1994.
Transient loss of serum protective activity following short-term stress: a
possible biochemical link between stress and atherosclerosis. Journal of Psy-
chosomatic Research 38, 871–884.
Arenz, C.L., 1997. Handling fox squirrels: ketamine-hydrochloride versus a simple
restraint. Wildlife Society Bulletin 25, 107–109.
Bao, A.M., Meynen, G., Swaab, D.F., 2008. The stress system in depression and
neurodegeneration: focus on the human hypothalamus. Brain Research
Reviews 57, 531–553.
Bauer, M.E., Perks, P., Lightman, S.L., Shanks, N., 2001. Restraint stress is associated
with changes in glucocorticoid immunoregulation. Physiology & Behavior 73,
T. Buynitsky, D.I. Mostofsky/Neuroscience and Biobehavioral Reviews 33 (2009) 1089–1098
in adult rats on behavior and hypothalamic-pituitary-adrenal responsiveness:
Comparison of two outbred rat strains. Behavioural Brain Research 172, 351–354.
Benatti, B.B., Nogueira-Filho, G.R., Diniz, M.C., Sallum, E.A., Nociti Jr., F.H., 2003.
Stress may enhance nicotine effects on periodontal tissues. An in vivo study in
rats. Journal of Periodontal Research 38, 351–353.
Bhat, M.S., Rao, G., Murthy, K.D., Bhat, P.G., 2007. Housing in pyramidal counteracts
neuroendocrine and oxidative stress caused by chronic restraint in rats. Evi-
dence-Based Complementary and Alternative Medicine 4, 35–42.
Bierhaus, A., Wolf, J., Andrassy, M., Rohleder, N., Humpert, P.M., Petrov, D., Ferstl, R.,
Von Eynatten, M., Wendt, T., Rudofsky, G., Joswig, M., Morcos, M., Schwaninger,
M., McEwen, B., Kirschbaum, C., Nawroth, P., 2003. A mechanism converting
psychosocial stress into mononuclear cell activation. Proceedings of the
National Academy of Sciences 100, 1920–1925.
Bizon, J.L., Helm, K.A., Han, J.S., Chun, H.J., Pucilowska, J., Kay Lund, P., 2001.
Hypothalamic-pituitary-adrenal axis function and corticosterone receptor
expression in behaviourally characterized young and aged long-evans rats.
European Journal of Neuroscience 14, 1739–1751.
Bosch, J.A., de Geus, J.C., Ligtenberg, T.J.M., Nazmi, K., Veerman, E.C.I., Hoogstaten, J.,
Nieuw Amergongen, A.V., 2000. Salivary MUC5B-mediated adherence (ex vivo)
of helicobacter pylori during acute stress. Psychosomatic Medicine 62, 40–49.
Bowman, R.E., Maclusky, N.J., Diaz, S.E., Zrull, M.C., Luine, V.N., 2006. Aged rats: Sex
differences and responses to chronic stress. Brain Research 1126, 156–166.
memory storage are reduced following a chronic stressful experience. Psycho-
pharmacology 129, 161–167.
Campan, P., Planchard, P.O., Duran, D., 1997. Pilot study on n-3 polyunsaturated
fattyacids inthetreatmentofhumanexperimentalgingivitis. JournalofClinical
Peridontology 24, 907–913.
Canu, M.H., Darnaude ´ry, M., Falempin, M., Maccari, S., Viltart, O., 2007. Effect of
hindlimb unloading on motor activity in adult rats: Impact of prenatal stress.
Behavioral Neuroscience 121, 177–185.
Ceballos, R.M., Faraday, M.M., Cousino Klein, L., 2006. Rat strain and sex differences
in leptin responses to immobilization stress. Pharmacology, Biochemistry and
Behavior 27, 136–146.
Cecchi, M., Khoshbouei, H., Morilak, D.A., 2002. Modulatory effects of norepinephr-
ine, acting on alpha1 receptors in the central nucleus of the amygdale, on
behavioral and neuroendocrine responses to acute immobilization stress. Neu-
ropharmacology 43, 1139–1147.
Chadda, R., Devaud, L.L., 2005. Differential effects of mild repeated restraint stress
on behaviors and GABAAreceptors in male and female rats. Pharmacology 81,
Chappell, P.B., Smith, M.A., Kilts, C.D., Bissette, G., Ritchie, J., Anderson, C., Nemeroff,
C.B., 1986. Alterations in corticotrophin-releasing factor-like immunoreactivity
in discrete rat brain regions after acute and chronic stress. The Journal of
Neuroscience 6, 2908–2914.
Chou-Green, J.M., Holscher, T.D., Dallman, M.F., Akana, S.F., 2003. Repeated stress in
young and old 5-HT2Creceptor knockout mice. Physiology & Behavior 79, 217–
Christian, L.M., Deichert, N.T., Goudin, J., Graham, J.E., Keicolt-Glaser, J.K., 2002.
Psychological influences on endocrine and immune function. Handbook of
Neuroscience for the Behavioral Sciences 3, 537–547.
Colomina, M.T., Sanchez, D.J., Sanchez-Turet, M., Domingo, J.L., 1999. Behavioral
effects of aluminum in mice: Influence of restraint stress. Neuropsychobiology
Conrad, C.D., Galea, L.A.M., Kuroda, Y., McEwen, B., 1996. Chronic stress impairs rat
spatial memory on the Y maze, and this effect is blocked by tianeptine pre-
treatment. Behavioral Neuroscience 6, 1321–1334.
Conrad, C.D., Magarin ˜os, A.M., LeDoux, J.E., McEwen, B.S., 1999. Repeated restraint
stress facilitates fear conditioning independently of causing hippocampal CA3
dendritic atrophy. Behavioral Neuroscience 5, 902–913.
Costa, A., Smeraldi, A., Tassorelli, C., Greco, R., Nappi, G., 2005. Effects of acute and
chronic restraint stress on nitroglycerin-induced hyperalgesia in rats. Neu-
roscience Letters 383, 7–11.
Dallman, M.F., 2007. Modulation of stress responses: how we cope with excess
glucocorticoids. Experimental Neurology 206, 179–182.
Dallman, M.F., Akana, S.F., Strack, A.M., Scribner, K.S., Pecoraro, N., la Fleur, S.E.,
Houshyar, H., Gomez, F., 2004. Chronic stress-induced effects of corticosterone
on brain: direct and indirect. Annals New York Academy of Sciences 1018, 141–
Dallman, M.F., Pecoraro, N., Akana, S.F., la Fleur, S.E., Gomez, F., Houshyar, H., Bell,
M.E., Bhatnagar, S., Laugero, K.D., Manalo, S., 2003. Chronic stress and obesity: a
new view of ‘‘comfort food. Proceedings of the National Academy of Sciences
Dallman, M.F., Pecoraro, N.C., la Fleur, S.E., 2005. Chronic stress and comfort foods:
self-medication and abdominal obesity. Brain, Behavior, and Immunity 19,
Dal-Zotto, S., Martı ´, O., Delgado, R., Armario, A., 2004. Potentiation of glucocorticoid
release does not modify the long-term effects of single exposure to immobi-
lization stress. Psychopharmacology 177, 230–237.
Davydov, V.V., Shvets, V.N., 2003. Age-dependent differences in the stimulation of
lipid peroxidation in the heart of rats during immobilization stress. Experi-
mental Gerontology 38, 693–698.
Derin, N., Aydin, S., Yargic ¸og ˘lu, P., Agar, A., 2006. Changes in visual evoked poten-
tials, lipid peroxidation and antioxidant enzymes in rats exposed to restraint
stress: effects of L-carntine. International Journal Neuroscience 116, 205–221.
Dhabhar, F.S., McEwen, B., Spencer, R.L., 1997. Adaptation to prolonged or repeated
stress-comparison between rat strains showing intrinsic differences in reactiv-
ity to acute stress. Neuroendocrinology 65, 360–368.
Echeverry, M.B., Guimara ˜es, F.S., Del Bel, E.A., 2004. Acute and delayed restraint
stress-induced changes in nitric oxide producing neurons in limbic regions.
Neuroscience 125, 981–993.
Engler, A., Roy, S., Sen, C.K., Padgett, D.A., Sheridan, J.F., 2005. Restraint stress alters
lung gene expression in an experimental influenza A viral infection. Journal of
Neuroimmunology 162, 103–111.
Fan, S.G., Shao, L., Ding, G.F., 1996. A suppressive protein generated in peripheral
lymph tissue induced by restraint stress. Advances in Neuroimmunology 6,
Freeman, M.L., Sheridan, B.S., Bonneau, R.H., Hendricks, R.L., 2007. Psychological
stress compromises CD8+ T cell control of latent herpes simplex virus type 1
infections. Journal of Immunology 179, 322–328.
Fujioka, T., Sakata, Y., Yamaguchi, K., Shibasaki, T., Kato, H., Nakamura, S., 1999. The
effects of prenatal stress on the development of hypothalamic paraventricular
neurons in fetal rats. Neuroscience 92, 1079–1088.
Furay, A.R., Bruestle, A.E., Herman, J.P., 2009. The role of the forebrain glucocortio-
coid receptor in acute and chronic stress. Endocrinology 149, 5482–5490.
Gagliano, H., Fuentes, S., Nadal, R., Armario, A., 2008. Previous exposure to immo-
dissociation between behavioral and pituitary-adrenal responses. Behavioural
Brain Research 187, 239–245.
Galea, L.A.M., McEwen, B.S., Tanapat, P., Deak, T., Spencer, R.L., Dhabhar, F.S., 1997.
Sex differences in dendritic atrophy of CA3 pyramidal neurons in response to
chronic restraint stress. Neuroscience 81, 689–697.
Gameiro, G.H., Gamiero, P.H., da Silva Andrade, A., Pereira, L.F., Arthuri, M.T.,
Marcondes, F.K., Ferraz de Arruda Veiga, M.C., 2006. Nociception- and anxi-
ety-like behavior in rats submitted to different periods of restraint stress.
Physiology & Behavior 87, 643–649.
Gas ˇpers ˇic ˇ, R., Sˇtiblar-Marinc ˇic ˇ, D., Skaleric ˇ, U., 2002. Influence of restraint stress on
ligature-induced periodontitis in rats. European Journal of Oral Science 110,
Gaerylewski, A., 2007. The trouble with animal models. The Scientist 21, 44–55.
Glaser, R., Kiecolt-Glaser, J.K., Marucha, P.T., MacCallum, R.C., Laskowski, B.F.,
Malarkey, W.B., 1999a. Stress-related changes in proinflammatory cytokine
production in wounds. Archives of General Psychiatry 56, 450–456.
Glaser, R., Rabin, B., Chesney, M., Cohen, S., Natelson, B., 1999b. Stress-induced
immunomodulation: implications for infectious diseases? Journal of the Amer-
ican Medical Association 281, 2268–2270.
Glavin, G.B., Pare ´, W.P., Sandbak, R., Bakke, H.K., Murison, R., 1994. Restraint stress
in biomedical research: an update. Neuroscience and Biobehavioral Reviews 18,
Glei, D.A., Goldman, N., Chuang, Y., Weinstein, W., 2007. Do chronic stressors lead to
physiological dysregulation? Testing the theory of allostatic load. Psychoso-
matic Medicine 69, 769–776.
by stress: implications for stress-related affective disorders. Neuroscience and
Biobehavioral Reviews 32, 1152–1160.
Grandin, T., Deesing, M., 2002. Distress in animals: is it fear, pain or physical stress?
American Board of Veterinary Practitioners.
Grandin, T., Curtis, S.E., Widowski, T.M., Thurmon, J.C., 1986. Electro-immobiliza-
tion versus mechanical restraint in an avoid-avoid choice test for ewes. Journal
of Animal Science 62, 1469–1480.
Grissom, N., Kerr, W., Bhatnagar, S., 2008. Struggling behavior during restraint is
regulated by stress experience. Behavioural Brain Research 191, 219–225.
Haas, D.A., George, S.R., 1988. Single or repeated mild stress increases synthesis and
release of hypothalamic corticotrophin-releasing factor. Brain Research 461,
Harvey, B.H., Bothma, T., Nel, A., Wegener, G., Stein, D.J., 2005. Involvement of the
NMDA receptor, NO-cyclic GMP and nuclear factor K-b in an animal model of
repeated trauma. Human Psychopharmacology: Clinical and Experimental 20,
Harvey, S.B., Wadsworth, M., Wessely, S., Hotopf, M., 2008. The relationship
between prior psychiatric disorder and chronic fatigue: evidence from a
national birth cohort study. Physiological Medicine 7, 933–940.
Hashimoto, M., Wantanabe, T., Fujioka, T., Tan, N., Yamashita, H., Nakamura, S., 2001.
Modulating effects of prenatal stress on hyperthermia induced in adult rat
offspring by restraint or LPS-induced stress. Physiology & Behavior 73, 125–132.
Head, C.C., Farrow, M.J., Sheridan, J.F., Padgett, D.A., 2006. Androstenediol reduces
the anti-inflammatory effects of restraint stress during wound healing. Brain,
Behavior & Immunity 20, 590–596.
Helm, H.A., Ziegler, D.R., Gallagher, M., 2004. Habituation to stress and dexametha-
sone suppression in rats with selective basal forebrain cholinergic lesions.
Hippocampus 14, 628–635.
Houri-Haddad, Y., Itzchaki, O., Ben-Nathan, D., Shapira, L., 2003. The effect of
chronic emotional stress on the humoral immune response to Porphyromonas
gingivalis in mice. Journal of Periodontal Research 38, 204–209.
Hsu, D.T., Chem, F.L., Takahashi, L.K., Kalin, N.H., 1998. Rapid stress-induced
elevations in corticotropin-releasing hormone mRNA in rat central amygdale
nucleus and hypothalamic paraventricular nucleus: an in situ hybridization
analysis. Brain Research 788, 305–310.
Hunzeker, J., Padgett, D.A., Sheridan, P.A., Dhabhar, F.S., Sheridan, J.F., 2004. Mod-
ulation of natural killer cell activity by restraint stress during an influenza A/
PR8 infection in mice. Brain, Behavior and Immunity 18, 526–535.
T. Buynitsky, D.I. Mostofsky/Neuroscience and Biobehavioral Reviews 33 (2009) 1089–1098
Inoue, T., Koyama, T., Muraki, A., Yamashita, I., 1993. Effects of single and repeated
immobilization stress on corticotrophin-releasing factor concentrations in
discrete rat brain regions. Progress in Neuro-Psychopharmacology & Biological
Psychiatry 17, 161–170.
Jackson, M.E., Moghaddam, B., 2006. Distinct patterns of plasticity in prefrontal
cortex neurons that encode slow and fast responses to stress. European Journal
of Neuroscience 24, 1702–1710.
Jeong, Y.H., Park, C.H., Yoo, J., Shin, K.Y., Ahn, S.M., Kim, H.S., Lee, S.H., Emson, P.C.,
Suh, Y.H., 2006. Chronic stress accelerates learning and memory impairments
and increases amyloid deposition in APPV717I-CT100 transgenic mice, an Alz-
heimer’s disease model. The FASEB Journal 20, 729–731.
Kang, J.E., Cirrito, J.R., Dong, H., Csernansky, J.G., Holtzman, D.M., 2007. Acute stress
increases interstitial fluid amyloid-b via corticotrophin-releasing factor and
neuronalactivity.Proceedings oftheNationalAcademyofSciences 104,10673–
Kesavalu, L., Bakthavatchalu, V., Rahman, M.M., Su, J., Raghu, B., Dawson, D.,
Fernandes, G., Ebersole, J.L., 2007. Omega-3 fatty acid regulates inflammatory
cytokine/mediator messenger RNA expression in Porphyromonas gingivalis-
induced experimental periodontal disease. Oral Microbiology Inmunology 22,
Khan, S., Liberzon, I., 2004. Topiramate attenuates exaggerated acoustic startle in an
animal model of PTSD. Psychopharmacology 172, 220–225.
Kiecolt-Glaser, J.K., Glaser, R., 2001. Psychological stress and wound healing.
Advances in Mind-Body Medicine 17, 15–16.
Kiecolt-Glaser, J.K., Marucha, P.T., Malarkey, W.B., Mercado, A.M., Glaser, R., 1995.
Slowing of wound healing by psychological stress. Lancet 346, 1194–1196.
King, C.D., Devine, D.P., 2007. Opioid modulation of reflex versus operant responses
following stress in the rat. Neuroscience 147, 174–182.
King, C.D., Devine, D.P., Vierck, C.J., Rodgers, J., Yezierski, R.P., 2003. Differential
effects of stress onescapeand reflex responses tonociceptivethermalstimuliin
the rat. Brain Research 987, 214–222.
Kitraki, E., Kremmyda, O., Youlatos, D., Alexis, M.N., Kittas, C., 2004. Gender-
dependent alterations in corticosteroid receptor status and spatial performance
following 21 days of restraint stress. Neuroscience 125, 47–55.
Kleen, J.K., Sitomer, M.T., Killeen, P.R., Conrad, C.D., 2006. Chronic stress impairs
spatial memory and motivation for reward without disrupting motor ability
and motivation to explore. Behavioral Neuroscience 120, 842–851.
Kumari, B., Kumar, A., Dhir, A., 2007. Protective effect of non-selective and selective
COX-2 inhibitors in acute immobilization stress-induced behavioral and bio-
chemical alterations. Pharmacological Reports 59, 699–707.
Early exposure to restraint stress enhances chemical carcinogenesis in liver.
Caner Letters 161, 215–220.
Lee, K., Kim, J., Seo, J., Kim, T., Im, J., Baek, I., Kim, K., Lee, J., Han, P., 2009. Behavioral
stress accelerates plaque pathogenesis in the brain of Tg2576 mice via gen-
eration of metabolic oxidative stress. Journal of Neurochemistry 108, 165–175.
Lehman, C.D., Rodin, J., McEwen, B., Brinton, R., 1991. Impact of environmental
stress on the expression of insulin-dependent diabetes mellitus. Behavioral
Neuroscience 105, 241–245.
Lemaire, V., Koehl, M., Le Moal, M., Abrous, D.N., 2000. Prenatal stress produces
learning deficits associated with an inhibition of neurogenesis in the hippo-
campus. Proceedings of the National Academy of Sciences 97, 11032–11037.
Lemaire, V., Lamarque, S., Le Moal, M., Piazza, P.V., Abrous, D.N., 2006. Postnatal
stimulation of the pups counteracts prenatal stress-induced deficits in hippo-
campus neurogenesis. Biological Psychiatry 59, 786–792.
Levine, S., 2005. Stress: an historical perspective. Handbook of Stress and the Brain
Li, L., Messas, E., Batista, E.L., Levine, R.A., Amar, S., 2002. Porphyromonas gingivalis
infection accelerates the progression of atherosclerosis in an apolipoprotein e-
deficient heterozygous murine model. Circulation 105, 861–867.
Liu, J., Mori, A., 1999. Stress, aging, and brain oxidative damage. Neurochemical
Research 24, 1479–1497.
Liu, J., Wang, X., Mori, A., 1994. Immobilization stress-induced antioxidant defense
changes in rat plasma: effect of treatment with reduced glutathione. Interna-
tional Journal of Biochemistry 26, 511–517.
Liu, J., Wang, X., Shigenaga, M.K., Yeo, H.C., Mori, A., Ames, B.N., 1996. Immobiliza-
tion stress causes oxidative damage to lipid, protein, and DNA in the brain of
rats. The FASEB Journal 10, 1532–1538.
Lucas, L.R., Wang, C.J., McCall, T.J., McEwen, B.S., 2007. Effects of immobilization
stress on neurochemical markers in the motivational system of the male rat.
Brain Research 1155, 108–115.
Luine, V., Villegas, M., Martinez, C., McEwen, B.S., 1994. Repeated stress causes
reversible impairments of spatial memory performance. Brain Research 639,
Lynch, W.J., Kushner, M.G., Rawleigh, J.M., Fiszdon, J., Carroll, M.E., 1999. The effects
of restraint stress on voluntary ethanol consumption in rats. Experimental and
Clinical Psychopharmacology 7, 318–323.
Maccari, S., Morley-Fletcher, S., 2007. Effects of prenatal restraint stress on the
hypothalamus-pituitary-adrenal axis and related behavioural and neurobiolo-
gical alterations. Psychoneuroendocrinology 32, S10–S15.
Madrigal, J.L.M., Moro, M.A., Lizasoain, I., Lorenzo, P., Ferna ´ndez, A.P., Rodrigo, J.,
Bosca ´, L., Leza, J.C., 2003. Induction of cuclooxygenase-2 accounts for restraint
stress-induced oxidative status in rat brain. Neuropsychopharmacology 28,
Magarin ˜os, A.M., McEwen, 1995a. Stress-induced atrophy of apical dendrites of
hippocampal CA3 neurons: comparison of stressors. Neuroscience 69, 83–88.
Magarin ˜os, A.M., McEwen, 1995b. Stress-induced atrophy of apical dendrites of
hippocampal CA3c neurons: involvement of glucocorticoid secretion and exci-
tatory amino acid receptors. Neuroscience 69, 89–98.
Masuda, J., Mitsushima, D., Kimura, F., 2004. Female rats living in small cages
respond to restraint stress with both adrenocortical corticosterone release and
acetylcholine release in hippocampus. Neuroscience Letters 358, 169–172.
McDougal, J.N., Marques, P.R., Burks, T.F., 1983. Restraint alters the thermic
response to morphine by postural interference. Pharmacology Biochemistry
& Behavior 18, 495–499.
McGuire, L., Heffner, K., Glaser, R., Needleman, B., Malarkey, W., Dickinson, S.,
Lemeshow, S., Cook, C., Muscarella, P., Melvin, W.S., Ellsion, E.C., Kiecolt-Glaser,
J.K., 2006. Pain and wound healing in surgical patients. Annals of Behavior
Medicine 31, 165–172.
McLaughlin, K.J., Gomez, J.L., Baran, S.E., Conrad, C.D., 2007. The effects of chronic
stress on hippocampal morphology and function: An evaluation of chronic
restraint paradigms. Brain Research 1161, 56–64.
Miyahara, S., Komori, T., Shizuya, K., Nomura, J., 1998. Changes of serum adenosine
deaminase activity induced by stress in rats. Human Psychopharmacology:
Clinical and Experimental 13, 325–327.
Munhoz,C., Madrigal, J.L.M.,Garcı ´a-Bueno, B., Pradillo, J.M.,Moro, M.A., Lizasoain, I.,
Lorenzo, P., Scavone, C., Leza, J.C., 2004. TNF-a accounts for short-term persis-
tence of oxidative status in rat brain after two weeks of repeated stress.
European Journal of Neuroscience 20, 1125–1130.
Nakajima, K., Hamada, N., Takahashi, Y., Sasaguri, K., Tsukinoki, K., Umemoto, T.,
Sato, S., 2006. Restraint stress enhances alveolar bone loss in an experimental
rat model. Journal of Periodontal Research 41, 527–534.
Pace, T.W.W., Gaylord, R., Topczewski, F., Girotti, M., Rubin, B., Spencer, R.L., 2005.
Immediate–early gene induction in hippocampus and cortex as result of novel
experience is not directly related to the stressfulness of that experience.
European Journal of Neuroscience 22, 1679–1690.
Pardon, M.C., 2007. Stress and ageing interactions: a paradox in the context of
shared etiological and physiopathological processes. Brain Research Reviews
Pardon, M.C., Rattray, I., 2008. What do we know aboutthe long-term consequences
of stress on ageing and the progression of age-related neurodegenerative
disorders? Neuroscience and Biobehavioral Reviews 32, 1103–1120.
Pare ´, W.P., Glavin, G.B., 1986. Restraint stress in biomedical research: a review.
Neuroscience and Biobehavioral Reviews 10, 339–370.
Pecoraro, N., Reyes, F., Gomez, F., Bhargava, A., Dallman, M., 2004. Chronic stress
promotes palatable feeding, which reduces signs of stress: feedforward and
feedback effects of chronic stress. Endocrinology 145, 3754–3762.
Pedersen, W.A., Culmseee, C., Ziegler, D., Herman, J.P., Mattson, M.P., 1999. Aberrant
stress response associated with severe hypoglycemia in a transgenic mouse
model of Alzheimer’s disease. Journal of Molecular Neuroscience 13, 159–165.
Popovic ´, M.,Jovana-Nes ˇic ´,K.,Popovic ´,N., 1996.Behavioral and adaptive status in an
experimental model of Alzheimer’s disease in rats. International Journal of
Neuroscience 86, 281–299.
Rissman, R.A., Lee, K.F., Vale, W., Sawchenko, P.E., 2007. Corticotropin-releasing
factor receptors differentially regulate stress-induced tau phosphorylation. The
Journal of Neuroscience 27, 6552–6562.
Rojas, I.-G., Padgett, D.A., Sheridan, J.F., Marucha, P.T., 2002. Stress induced suscept-
ibility to bacterial infection during cutaneous wound healing. Brain, Behavior,
and Immunity 16, 74–84.
Rossi-George, A., Virgolini, M.B., Weston, D., Cory-Slechta, D.A., 2008. Alterations in
glucocorticoid negative feedback following maternal Pb, prenatal stress and the
combination: a potential biological unifying mechanism for their correspond-
ing disease profiles. Toxicology and Applied Pharmacology.
Ruys, J.D., Mendoza, S.P., Capitanio, J.P., Mason, W.A., 2004. Behavioral and phy-
siologicaladaptation torepeatedchair restraintin rhesusmacaques. Physiology
& Behavior 82, 205–213.
S ¸ahin, E., Gu ¨mu ¨s ¸lu ¨, S., 2007. Immobilization stress in rat tissues: Alterations in
protein oxidation, lipid peroxidation and antioxidation defense system. Com-
parative Biochemistry and Physiology, Part C 144, 342–347.
Sapolsky, R., 1982. The endocrine stress-response and social status in the wild
baboon. Hormones and Behavior 16, 279–292.
Sapolsky, R.M., Romero, M., Munck, A.U., 2000. How do glucocorticoids influence
stress responses integrating permissive, suppressive, stimulatory, and prepara-
tive actions. Endocrine Reviews 21, 55–89.
Sarkar, S., Zaretskaia, M.V., Zaretsky, D.V., Moreno, M., DiMicco, J.A., 2007. Stress-
and lipopolysaccharide-induced c-fos expression and nNOS in hypothalamic
neurons projecting to medullary raphe in rats: a triple immunofluorescent
labeling study. European Journal of Neuroscience 26, 2228–2238.
Schoenfeld, W.N., 1981. Pain: a verbal response. Neuroscience and Biobehavioral
Reviews 5, 385–389.
Seligman, M., 1975. Helplessness: On Depression, Development, and Death. W.H.
Freeman, San Francisco.
Selye, H., 1976. Forty years of stress research: principal remaining problems and
misconceptions. Canadian Medical Association Journal 115, 53–56.
Seo,Y.J.,Kwon, M.S.,Shim,E.J., Park,S.H.,Choi,O.S.,Suh, H.W.,2006.Changesin pain
behavior induced by formalin, substance P, glutamate and pro-inflammatory
cytokines in immobilization-induced stress mouse model. Brain Research
Bulletin 71, 279–286.
Sheridan, J.F., Padgett, D.A., Avitsur, R., Marucha, P.T., 2004. Experimental models of
stress and wound healing. World Journal of Surgery 28, 327–330.
Sotiropoulos, I., Cerqueira, J.J., Catania, C., Takashima, A., Sousa, N., Almeida, O.F.X.,
2008. Stress and glucocorticoid footprints in the brain – the path from depres-
T. Buynitsky, D.I. Mostofsky/Neuroscience and Biobehavioral Reviews 33 (2009) 1089–1098
sion to Alzheimer’s disease. Neuroscience and Biobehavioral Reviews 32, 1161–
Strausbaugh, H.J., Dallman, M.F., Levine, J.D., 1999. Repeated, but not acute, stress
suppresses inflammatory plasma extravasation. Proceedings of the National
Academy of Sciences 96, 14629–14634.
Swaab, D.F., Bao, A.M., Lucassen, P.J., 2005. The stress system in the human brain in
depression and neurodegeneration. Ageing Research Reviews 4, 141–194.
Tabarin,A.,Diz-Chaves, Y.,Consoli,D., Monsaingeon, M., Bale, T.L.,Culler,M.D., Datta,
R., Drago, F., Vale, W.W., Koob, E.P., Zorrilla, E.P., Contarino, A., 2007. Role of the
corticotrophin-releasing factor receptor type 2 in the control of food intake in
mice: a meal pattern analysis. European Journal of Neuroscience 26, 2303–2314.
Taylor, B.K., Peterson, M.A., Basbaum, A.I., 1995. Exaggerated cardiovascular and
behavioral nociceptive responses to subcutaneous formalin in the spontaneous
hypertensive rat. Neuroscience Letters 201, 9–12.
van Rensburg, S.J., van Zyl, J.M., Potocnik, F.C.V., Daniels, W.M.U., Uys, J., Marais, L.,
Hon, D., van der Walt, B.J., Erasmus, R.T., 2006. The effect of stress on the
antioxidative potential of serum: implications for Alzheimer’s disease. Meta-
bolic Brain Disease 21, 171–179.
Vyas, A., Mitra, R., Shankaranarayana, R.B.S., Chattarji, S., 2002. Chronic stress
induces contrasting patterns of dendritic remodeling in hippocampal and
amygdaloid neurons. Journal of Neuroscience 22, 6810–6818.
Wang, J., Charboneau, R., Barke, R.A., Loh, H.H., Roy, S., 2002. m-Opioid receptor
mediates chronic restraint stress-induced lymphocyte apoptosis. The Journal of
Immunology 169, 3630–3636.
Weinberg, M.S., Girotti, M., Spencer, R.L., 2007. Restraint-induced fra-2 and c-fos
expression in the rat forebrain: Relationship to stress duration. Neuroscience
Wheway, J., Herzog, H., Mackay, F., 2007. NPY and receptors in immune and
inflammatory diseases. Current Topics in Medicinal Chemistry 7, 1743–
Wood,G.E.,Young, L.T., Reagan, L.P., McEwen,B.S.,2003. Acuteand chronic restraint
stress alter the incidence of social conflict in male rats. Hormones and Behavior
Wu, J., Song, T.-B., Li, Y.-J., He, K.-S., Ge, L., Wang, L.-R., 2007. Prenatal restraint stress
impairs learning and memory and hippocampal PKCbeta1 expression and
translocation in offspring rats. Brain Research 1141, 205–213.
Yaras, N., Yargicoglu, P., Agar, A., Gumuslu, S., Abidin, I., Ozdemir, S., 2003. Effect of
immobilization and cold stress on visual evoked potentials. International
Journal Neuroscience 113, 1055–1067.
Yehuda, S., Rabinovitz, S., Carasso, R.L., Mostofsky, D.I., 2008. Long-lasting cognitive,
physiological and hematological effects in rehabilitated, early dietary iron-
deficiency adult rats, and improvement by treatment with a mixture of essen-
tial fatty acids. Nutritional Neuroscience 11, 167–171.
Yoshihara, T., Yawaka, Y., 2008. Repeated immobilization stress in the early post-
natal period increases stress response in adult rats. Physiology & Behavior 93,
Zafir, A., Banu, N., 2008. Modulation of in vivo oxidative status by exogenous
corticosterone and restraint stress in rats. Stress 12, 167–177.
Zhou, Y., Elkins, P.D., Howell, L.A., Ryan, D.H., Harris, R.B.S., 1998. Apolipoprotein-E
deficiency results in an altered stress responsiveness in addition to an impaired
spatial memory in young mice. Brain Research 788, 151–159.
Zorzet, S., Perissin, L., Rapozzi, V., Giraldi, T., 1998. Restraint stress reduces the
and Immunity 12, 23–33.
T. Buynitsky, D.I. Mostofsky/Neuroscience and Biobehavioral Reviews 33 (2009) 1089–1098