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The Journal of Neuroscience, March 15, 2003, 23(6):2453
A Critical Role for the Cannabinoid CB1 Receptors in
Alcohol Dependence and Stress-Stimulated Ethanol Drinking
Ildiko
Racz1,
Andras
Bilkei-Gorzo1,
Zsuzsanna E.
Toth2,
Kerstin
Michel1,
Miklós
Palkovits2, and
Andreas
Zimmer1
1 Laboratory of Molecular Neurobiology, Department of
Psychiatry, University of Bonn, 53105 Bonn, Germany, and
2 SE Laboratory: Laboratory of Neuromorphology, Department
of Anatomy, Faculty of Medicine, Semmelweis University, 1094 Budapest
Tüzolto u.58., Hungary
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ABSTRACT |
Although many people drink alcohol regularly, only some become
addicted. Several studies have shown that genetic and environmental factors contribute to individual differences in the vulnerability to
the effects of alcohol (Nestler, 2000 ; Kreek, 2001; Crabbe, 2002).
Among the environmental factors, stress is perhaps the most important
trigger for relapse after a period of abstinence (Koob and Nestler,
1997; Piazza and Le Moal, 1998; Koob and Le Moal, 2001; Weiss et al.,
2001). Here we show that ethanol withdrawal symptoms were completely
absent in cannabinoid CB1 receptor-deficient mice, although
acute effects of ethanol and ethanol tolerance and preference were
basically normal. Furthermore, foot-shock stress had no affect on
alcohol preference in Cnr1 / mice, although it
induced a dramatic increase in Cnr1+/+ animals.
These results reveal a critical role for the CB1 receptor in clinically important aspects of alcohol dependence and provide a
rationale for the use of CB1 receptor antagonists in the
treatment of alcohol addiction.
Key words:
cannabinoid; ethanol; mice; mutation; withdrawal; addiction
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Introduction |
Three lines of evidence point to a
possible involvement of the cannabinoid
CB1 receptor in ethanol effects. First,
although the mechanisms of action of ethanol and
 9-tetrahydrocannabinol (THC), the natural CB1
agonist found in Cannabis sativa preparations, are
different, they produce a number of similar physiological and
behavioral responses, including euphoria, motor incoordination, and
hypothermia (Hungund and Basavarajappa, 2000a). Second, alcohol preference and self-administration can be modulated with
CB1 receptor agonists and antagonists (Colombo et
al., 1998; Gallate et al., 1999; Rodriguez de Fonseca et al., 1999;
Hungund and Basavarajappa, 2000a; Lallemand et al., 2001). Finally,
Buck et al. (1997) have identified a marker locus (D4Ncvs78) associated
with alcohol withdrawal liability on chromosome 4 in close proximity to Cnr1.
We therefore asked whether a deletion of the CB1
receptor would alter behavioral or physiological effects of alcohol.
Chronic ethanol exposure selectively increased the synthesis of
endocannabinoids in cell cultures (Basavarajappa and Hungund, 1999b;
Basavarajappa et al., 2000) and in mouse brains (Hungund and
Basavarajappa, 2000a). In addition, chronic ethanol treatment resulted
in a reduction of CB1 receptor densities and a
concomitant decrease in Bmax without any change in G-protein affinity Kd
(Basavarajappa et al., 1998; Basavarajappa and Hungund, 1999a). The
downregulation of CB1 receptors parallels in many
aspects the changes in CB1 expression and
signaling properties after chronic treatment with natural or synthetic
CB1 agonists, including anandamide (Rodriguez de
Fonseca et al., 1994; Basavarajappa and Hungund, 1999a). These
homeostatic adaptations of the endocannabinoid system may therefore
contribute to many of the physiological and behavioral effects of
chronic ethanol exposure, including tolerance and dependence. Indeed,
cross-tolerance between ethanol and THC have been reported from many
studies in the literature (Hungund and Basavarajappa, 2000a). If this
hypothesis were correct, one would expect to see alterations in the
development of tolerance and dependence in
CB1-deficient mice.
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Materials and Methods |
Acute ethanol effects, tolerance, and withdrawal.
Six- to 8-week-old male Cnr1+/+ and
Cnr1/ mice, with a C57BL/6J genetic
background, were used. Animals were housed individually under reversed
light-dark conditions (lights on at 7:00 P.M. and lights off at 9:00
A.M.). To determine acute alcohol effects, animals received a single
intraperitoneal injection of vehicle (PBS) and 1, 2, or 4 gm/kg
ethanol. Body temperatures were measured with a rectal thermometer
immediately before and 30 min after the treatment. Another group of
animals was trained on a rotarod (4-25 rpm with an acceleration of 1 rpm/sec; Columbus Instruments, Columbus, OH) 10 times
daily for 2 d. Animals that did not reach the training criterion
( 30 sec on the rotating rod without falling down, three times
successively) were not included in the test. On the third day, the
animals received a vehicle or ethanol (2 or 4 gm/kg) injection. Thirty
minutes after treatment, we measured the time the animals remained on
the rotating rod. To assess ethanol tolerance, animals were supplied
with ethanol solutions as their only drinking source as follows: days
1-3, 4% ethanol; days 4-10, 8% ethanol; and days 11-21, 16%
ethanol. Ethanol consumption (in grams per kilogram), food
consumption, and the body weight were recorded twice per week.
After 2 weeks of 16% ethanol drinking, the effect of acute ethanol
treatment on the body temperature of the animals was determined again.
The treatment procedure was the same as described above. For withdrawal studies, animals received the 16% ethanol solution for 3 more weeks
(days 11-41) before it was replaced with water on day 42. Quantification of withdrawal symptoms was made by a person, who was
blind to the experimental groups, using handling-induced convulsions performed as described previously (Watson et al., 1994) 3 hr after replacing ethanol with water. The behavioral ratings in response to
gentle handling during ethanol withdrawal were as follows: 0, no tremor
or convulsion; 1, mild tremor on lifting and turning; 2, continuous
severe tremor on lifting and turning; and 3, clonic forelimb extensor
spasm on lifting. For statistical analysis, the mean value and
SE of the body temperature and of the time the mice spent on the
rotarod were calculated. Groups were compared by two-way ANOVA
(genotype × treatment), followed by Scheffe post hoc
test. The differences between scores were calculated by nonparametric ANOVA, with the Kolmogorov-Smirnov test.
Open-field test. Mice were placed into the center of the
open-field apparatus (44 × 44 × 30 cm; Med
Associates, Georgia, VT) during the drinking procedure
and 3 d after the withdrawal. Movements of the animals were
tracked by an automatic monitoring system (Med Associates)
for 10 min. Horizontal motor (distance traveled) and central activity
(distance traveled in central area/total distance traveled) was
evaluated. The experiment was performed under low-light conditions
(~5 lux). Mean value and SE was calculated in each group, which
contained 10 animals. Groups were compared by two-way ANOVA
(genotype × treatment), followed by the Fisher's test.
Elevated zero maze. Animals were treated with saline or
ethanol (2 gm/kg) intraperitoneally in the volume of 10 ml/kg; 9-10 animals were tested in each group. Thirty minutes later, their activity
on the zero maze was measured for 5 min. The maze consisted of an
annular white platform (inner diameter of 46 cm, 5.6 cm width) elevated
40 cm above the ground level and equally divided into four quadrants.
The two opposite quadrants were enclosed by white walls (24 cm high) on
both edges of the platform. The behavior of mice was videotaped using a
camera fixed above the maze and analyzed with a video-tracking system
(Videomot; TSE Systems, Bad Homburg, Germany). The number of stretching
postures was determined by an experienced observer unaware to strain or treatment. Time spent in the open area, distance traveled in the open
and closed parts, and number of stretching postures were evaluated
(Shepherd et al., 1994; Konig et al., 1996). Mean value and SE was
calculated in each group, and groups were compared by one-way ANOVA,
followed by the Fisher's test
Alcohol preference. Ethanol preference measurements were
basically performed as described previously (Little et al., 1999). Briefly, two drinking bottles (with a metal ball in the sipper tubes to
stop the dropping of fluids; Cascade 5; Hagen, Holm, Germany)
were available to the animals during the experiment. One of these
bottles contained 8% v/v alcohol, and the other contained drinking
water. The positions of the bottles were changed daily. The ratio of
alcohol to total fluid consumption, the amount of consumed ethanol (in
grams per kilogram), the body weight, and the food consumption were
determined twice per week.
Foot-shock procedure. This procedure was made with the
animals that had access to an ethanol solution (8%) ad
libitum for >5 weeks and maintained a stable ethanol intake. For
the foot-shock stress, animals were kept in a dark chamber during the
shock procedure, where a continuous background white noise (65 dB) was
present. A few seconds before the shock, a warning signal (sound and
light) was presented. Intermittent electric foot shocks (intensity, 0.5 mA; duration, 100 msec; interval between shocks, 55-60 sec) were then
delivered five times through the grid floor by an isolated stimulator.
The ratio of alcohol to total fluid consumption and the amount of
consumed ethanol (in grams per kilogram) was determined 24 and 96 hr
after the shock.
c-fos expression. Three brains from each
experimental group were analyzed. Brain sections (10 µm) were cut
using a cryostat and thaw mounted on Superfrost Plus glass slides
(Fisher, Pittsburgh, PA). After drying, the slide-mounted sections were
stored at  70°C. The template for c-fos was a murine cDNA
(400 bp). Plasmid was linearized to generate either sense or antisense
cRNA probes. In situ hybridization was performed as
described previously (Campbell and Hess, 1999). Hybridized sections
were covered with Kodak NTB emulsion (Eastman Kodak,
Rochester, NY) and exposed for 4 weeks. After development, the
background were stained in 0.5% Giemsa (Fluka, Neu-Ulm,
Germany), and the sections were dried and covered with Cytoseal 60 (Richard-Allan Scientific, Kalamazoo, MI).
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Results |
We first evaluated ethanol-induced hypothermia and
motor-incoordination after a single intraperitoneal injection of
ethanol solutions. As shown in Figure
1A, ethanol produced a
similar dose-dependent reduction in body temperature in mutant and
wild-type animals (treatment effect,
F(3,73) = 195.97, p < 0.0001; genotype effect, F(1,73) = 0.57, p = 0.45). In addition, motor coordination, as evaluated on the rotarod, was similarly affected by the ethanol treatment in both genotypes (treatment effect,
Cnr1+/+,
F(2,36) = 64.8, p  0.0001; Cnr1/,
F(2,35) = 79.6, p 0.0001; treatment × strain,
F(2,71) = 2.39, p = 0.0994), although Cnr1/ mice did not
perform as well in this test as Cnr1+/+
animals (genotype effect: F(1,71) = 7.6, p 0.01). Anxiolytic effects of subchronic
ethanol treatment (2 gm/kg, i.p.) were determined in the zero-maze
test. Animals of both genotypes spent significantly more time in the
open sectors after ethanol treatment and showed a reduced number of
stretch-attend postures. Thus, the anxiolytic properties of ethanol
were not affected by the mutation. (treatment × genotype,
F = 0.09, p = 0.767). These results
show that the CB1 receptor is not required for
these acute ethanol effects.
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Figure 1.
Acute ethanol effects and tolerance are identical
in wild-type and CB1 receptor-deficient mice.
A, Cnr1+/+ and
Cnr1/ mice showed a similar dose-dependent
reduction in body temperature and impairment of motor coordination on
the rotarod after intraperitoneal injection of ethanol that was
significant at 2 and 4 gm/kg. Interestingly, Cnr1+/+
animals performed better in the rotarod test than
Cnr1/ mice (genotype effect,
F(1,71) = 7.6, p = 0.0074). This is in contrast to our previous analysis of the mutant
phenotype on a mixed (129 × C57BL/6J) genetic background, in
which mutant animals showed a tendency toward a reduced running time
but never performed significantly different from wild-type controls
(Steiner et al., 1999). B, Anxiolytic effects of ethanol
were similar in mice from both genotypes. *p 0.05; **p < 0.001. When animals were forced to
drink an ethanol solution for a period of 3 weeks (4-16%), before
receiving an ethanol injection, only the highest dose tested (4 gm/kg)
produced a significant reduction in body temperature.
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We next wanted to examine the development of tolerance after chronic
ethanol exposure. For this purpose, we restricted the animals to an
ethanol solution (4-16%) as their only fluid source for a period of 3 weeks. The total liquid intake of
Cnr1/ and
Cnr1+/+ animals was similar at ~2-2.5
ml/d. Subsequently, we determined the physiological effects of an
intraperitoneal ethanol injection by measuring the animals' body
temperature. Only the highest dose of 4 gm/kg ethanol produced
significant hypothermia in these mice (Fig. 1C), whereas 2 gm/kg were already effective in alcohol-naive animals. However, there
was no difference between the two genotypes (treatment effect:,
F(3,28) = 32.05, p < 0.0001; genotype effect, F(1,28) = 0.52, p = 0.48; treatment × genotype,
F(3,1) = 0.747, p = 0.526) and, thus, ethanol tolerance was not affected by the CB1 deletion.
To study ethanol withdrawal symptoms, we restricted mice to a 16%
ethanol solution as their only fluids source for 4 weeks and then
replaced the ethanol solution with water. Withdrawal symptoms were
evaluated 3 hr after replacing the ethanol solution. Although
Cnr1+/+ animals displayed severe
withdrawal symptoms (mean score, 1.74 ± 0.19;
 2 = 15.2; p = 0.001;
Kolmogorov-Smirnov test), we could not detect any signs for ethanol
withdrawal in Cnr1/ animals (mean
score, 0.46 ± 0.12; 2 = 4.2;
p = 0.24; Kolmogorov-Smirnov test) (Fig.
2). We also measured withdrawal-induced
hyperlocomotion in the open field, which is a different symptom for
ethanol withdrawal in mice. As expected, Cnr1+/+ animals were significantly
hyperactive 3 d after ethanol withdrawal (F(1,17) = 7.10; p = 0.016; ANOVA). However, there was no change in
Cnr1/ mice
(F(1,11) = 1.00; p = 0.333; ANOVA). Thus, ethanol withdrawal symptoms were completely absent
in Cnr1/ mice.
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Figure 2.
Ethanol withdrawal symptoms are absent in
Cnr1/ mice. Animals had access to a 16% ethanol
solution as their only drinking source for 4 weeks.
Cnr1+/+ animals displayed severe withdrawal symptoms
3 hr after replacing the ethanol solution with water, whereas
Cnr1/ animals did not display any withdrawal
symptoms. ***p 0.005; Kolmogorov-Smirnov
test.
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CB1 receptor agonists and antagonists are known
modulators of appetite, food intake, and ethanol preference (Ameri,
1999). We therefore asked whether the preference for ethanol would be altered in the absence of CB1 receptors. When
animals were given access ad libitum to either an ethanol
solution (8%) or water, Cnr1/ mice
initially showed a significantly higher preference for the ethanol
solution than Cnr1+/+ animals (Fig.
3A). The absolute
amount of ethanol consumed was also higher in
Cnr1/ mice (B).
However, both genotypes established a similar stable level of ethanol
intake within a few days. After the first week, the
Cnr1/ animals showed the same ethanol
preference as Cnr1+/+ mice, and they
consumed a similar amount of ethanol until the end of the experiment.
There was no difference between the two genotypes in the amount of food
consumed, nor was there any genotype difference in the body weight
(data not shown).
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Figure 3.
Ethanol drinking behavior in a two-bottle choice
paradigm. A, Cnr1/ mice initially
showed a significantly higher preference for the ethanol (8%) solution
than Cnr1+/+ animals. B, The absolute
amount of ethanol consumed was also higher. Animals of both genotypes
established a similar stable level of ethanol intake after the first
week. *p 0.05; ANOVA.
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Stress is one of the most important factors known to trigger relapse
after a period of abstinence in human patients and in animal models of
drug addiction. In rodents, a brief exposure to a mild foot-shock
stressor can reinstate drug-seeking behavior or increase ethanol
preference. We therefore exposed mice that had access ad
libitum to an ethanol solution (8%) for >5 weeks and maintained
a stable ethanol intake to a mild 5 min foot shock. As expected,
wild-type Cnr1+/+ mice drank more ethanol
in the 24 hr period after receiving the foot shocks and also displayed
a significant increase in their preference for ethanol during this
period (Fig. 4). The stress-induced increase in ethanol preference was transient, because animals returned
to prestress levels within 96 hr. In contrast, however, ethanol
preference or absolute amount of ethanol consumed by
Cnr1/ mice were totally unaffected by
the foot-shock stressor.
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Figure 4.
Absence of stress-induced increase of ethanol
drinking in Cnr1/ mice. Animals had access
ad libitum to ethanol for a period of 5 weeks and
reached a stable plateau of ethanol consumption. A,
left and right, When exposed to a foot-shock
stress, Cnr1+/+ control animals displayed a
significant transient increase in ethanol preference and in the
absolute amount of ethanol consumed. In contrast,
Cnr1/ mice were not affected by the foot-shock
stressor. B, Jump responses elicited by the foot shocks
were similar in both genotypes. *p 0.05; ANOVA;
**p < 0.001.
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We considered the possibility that the level of stress produced by the
foot shock induced was different between the two genotypes. We
therefore evaluated the amplitude of jump responses after the administration of foot shocks. As shown in Figure 4B,
there was no difference in this parameter between
Cnr1+/+ and
Cnr1/ mice, indicating that the
immediate level of discomfort was similar. We also determined the level
of c-fos induction after the foot-shock stressor in
different brain regions (Fig. 5).
Cnr1+/+ and
Cnr1/ mice showed a robust
c-fos expression in the cortex, amygdala, hippocampus, and
paraventricular-thalamic nuclei in stressed, but not in control
animals. Together these results strongly indicate that the foot-shocks
produced similar levels of stress in
Cnr1/ and
Cnr1+/+ animals.
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Figure 5.
Stress-induced c-fos induction.
Representative sections through different brain regions are shown.
Cnr1+/+ and Cnr1/ mice showed
similar levels of c-fos induction in the cortex,
amygdala, hippocampus, and thalamus.
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Discussion |
Mice with specific gene deletions have been used recently to
investigate the role of the endocannabinoid system in drug
reinforcement and addiction. In this manuscript, we analyzed ethanol
responses in a mouse strain with a deletion in the cannabinoid
CB1 receptor gene Cnr1. Our results demonstrate a
crucial role of the CB1 receptor in the
physiological manifestation of ethanol dependence and in stress-induced
increase of ethanol preference.
The Cnr1 mutation was examined on a genetic C57BL/6J genetic
background. Mice with this genetic background are known for their voluntary consumption of alcoholic solutions (McClearn and Rodgers, 1961; Crabbe and Belknap, 1980). Unlike mice from many other genetic backgrounds, C57BL/6J animals will readily drink ethanol solutions when
these are presented together with regular tap water in a two-bottle
choice paradigm. Surprisingly, we found that
Cnr1/ mice had initially an even
higher preference for ethanol than C57BL/6J mice. This result was
unexpected, because previous pharmacological studies suggested that
blocking the CB1 receptor with the selective antagonist SR141716A reduced ethanol consumption. However, the interpretation of results obtained with this compound have been difficult, because SR141716A has a well known reverse agonist activity.
In addition, [3H]GTP S binding studies
on Cnr1/ brain tissues suggested that
SR141716A may have activity on a still unidentified "CB3" receptor
(Breivogel et al., 2001). Of course, it is also possible that the acute
blockade of the receptor produces different effects than the continuous
removal through the genetic ablation. In fact, a recent study has
revealed differential effects of SR141716A treatment of ethanol
preference, depending on the treatment regimen (Lallemand et al.,
2001). The increased preference of
Cnr1/ mice for ethanol was only
significant during the first days. After 1 week, the ethanol
consumption was virtually identical between
Cnr1+/+ and
Cnr1/ mice. Together, these results
indicate that the endocannabinoid system is not a critical mediator of
normal alcohol drinking behavior, although it may modulate behavioral
responses to the first contact with ethanol.
A completely different picture emerged when mice, which had established
a stable plateau of ethanol drinking, were exposed to a mild
intermittent foot-shock stress. As expected, alcohol consumption
increased significantly in Cnr1+/+ mice
for a brief period after the stress, whereas
Cnr1/ mice showed no change in their
ethanol preference. We can exclude the possibility that the foot shock
was less stressful for Cnr1/ animals,
because (1) Cnr1/ and
Cnr1+/+ mice exhibited similar jumping
responses and (2) c-fos induction through the foot-shock
stress was similar in both genotypes. Furthermore, two
Cnr1/ animals, but none of the
Cnr1+/+ mice, died shortly after the
foot-shock stress. We often noticed an increased mortality in
Cnr1/ mice (Zimmer et al., 1999),
especially when circumstances dictated higher stress levels (e.g., tape
testing for pinworm infections and construction work in the animal
facility). The cause of the deaths of these animals remains unclear but
may involve epileptic seizures. Previous studies using an independently
derived Cnr1/ strain on an outbred
genetic background have also indicated that Cnr1/ mice are more emotional than
Cnr1+/+ animals (Martin et al., 2002),
although these animals exhibited reduced analgesia after a swim stress
(Valverde et al., 2000). In addition, the acquisition and consolidation
of conditioned auditory freezing behaviors after foot-shock stimulation
was unaltered in a third independent Cnr1 knock-out strain
(Marsicano et al., 2002).
Numerous clinical studies have revealed a general correlation between
stress and drug relapse. Stressful life events in childhood (Simantov
et al., 2000), daily job problems in adulthood (Delaney et al., 2002),
and experiences like the September 11th terror attack (Vlahov et al.,
2002) each may increase the risk for alcohol drinking and a
concomitantly increased risk for alcoholism. Reinstatement of
drug-seeking behaviors through intermittent foot-shock stress has been
demonstrated for a number of drugs, including heroin (Shaham and
Stewart, 1995), cocaine (Erb et al., 1996), nicotine (Buczek et al.,
1999), and alcohol (Le et al., 1998), suggesting a drug-independent
effect of the stressor. Although the molecular and cellular
events underlying stress-induced reinstatement are primarily obscure,
corticotropin-releasing factor, as well as dopamine-dependent
and dopamine-independent mechanisms, have been implicated (Self and
Nestler, 1998; Le and Shaham, 2002; Leri et al., 2002). There is almost
no previous evidence for a genetic predisposition to stress-induced
relapse, except for one study that demonstrated enhanced stress-induced
ethanol drinking in corticotrophin-releasing hormone-1
receptor-deficient mice (Sillaber et al., 2002).
Our second important finding is the demonstration that withdrawal
symptoms after the cessation of chronic ethanol administration were
completely absent in CB1 knock-out mice. To our
knowledge, this is the first direct evidence for an involvement of the
endocannabinoid system in ethanol withdrawal. Importantly, however, the
analysis of recombinant inbred strains for ethanol withdrawal
severity lead to the identification of a quantitative trait locus
(QTL) on chromosome 4 in close proximity to Cnr1 (Buck et al.,
1997), which may be independent from another distal locus on the same chromosome (Fehr et al., 2002). It is well known that
CB1 receptor densities and
CB1 receptor agonist-stimulated
[35S]GTPS binding differ
significantly between the parental alcohol-preferring C57BL/6 and
alcohol-avoiding DBA/2J strains (Hungund and Basavarajappa, 2000b;
Basavarajappa and Hungund, 2001). Therefore, it seems possible that the
Cnr1 locus accounts for this QTL. Interestingly, a recent clinical
study also associated a Cnr1 gene polymorphism with the severity of
withdrawal symptoms in humans (Schmidt et al., 2002).
Ethanol withdrawal symptoms may involve homeostatic changes of the
endocannabinoid system, including an increased synthesis of
endocannabinoids and a concomitant downregulation of
CB1 receptor binding sites (Basavarajappa and
Hungund, 2002; Gonzalez et al., 2002). The endocannabinoid system may
play a general role in the manifestation of physiological drug
dependence, because a significant reduction of withdrawal symptoms was
also observed in CB1-deficient mice after
morphine withdrawal (Ledent et al., 1999).
In summary, the endocannabinoid system does not seem to be crucial for
the rewarding effects of ethanol and the manifestation of normal
ethanol drinking behaviors. However, it appears to play an important
role in the manifestation of stress-induced alcohol drinking and
ethanol withdrawal. These results support the notion that the neuronal
mechanisms involved in drug reinforcement are dissociable from those
involved in withdrawal and reinstatement (Shalev et al., 2002). Indeed,
our results demonstrate that ethanol tolerance and physical dependence
can be separated. Thus, CB1 receptor antagonists
may be useful for the treatment of alcohol addiction.
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FOOTNOTES |
Received Nov. 5, 2002; revised Dec. 13, 2002; accepted Dec. 23, 2002.
This work was supported by grants from the Land Nordrhein Westphalen
(Innovationsprogramm Forschung), the Deutsche Forschungsgemeinschaft (Sonderforschungsbereich 400), and the German National Genome Network. We thank members of the laboratory for suggestions on the
experiments and this manuscript.
Correspondence should be addressed to Andreas Zimmer, Laboratory of
Molecular Neurobiology, Clinic of Psychiatry, University of Bonn,
Sigmund-Freud-Strasse 25, 53125 Bonn, Germany. E-mail: neuro{at}uni-bonn.de.
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TOP
ABSTRACT
Introduction
