Vous êtes sur la page 1sur 7

Psychopharmacology (2004) 172:264270

DOI 10.1007/s00213-003-1647-z

ORIGINAL INVESTIGATION

Cristina Orsini Francesca Buchini


Pier Vincenzo Piazza Stefano Puglisi-Allegra
Simona Cabib

Susceptibility to amphetamine-induced place preference is predicted


by locomotor response to novelty and amphetamine in the mouse
Received: 28 February 2003 / Accepted: 4 September 2003 / Published online: 4 November 2003
 Springer-Verlag 2003

Abstract Rationale: It has been demonstrated that major


differences between mice of the C57BL/6J and DBA/2J
inbred strains for amphetamine-induced place conditioning (preference and avoidance, respectively) are evident
in standard housing conditions but abolished by temporary restricted feeding. This gene-experience model may
be usefully exploited to dissect behavioral phenotypes
related to place conditioning induced by addictive drugs.
Objectives: This study evaluated a number of behavioral
phenotypes related to amphetamine-induced place preference for strain differences (C57BL/6J vs DBA/2J)
susceptible to be abolished by temporary food restriction.
Methods: Mice of the two inbred strains were tested for:
(1) conditioned taste aversion and place preference
induced by amphetamine within the same dose-range;
(2) preference for a novel compartment 24 h after a single
exposure to only one of two compartments; (3) amphetamine-induced behavioral sensitization and conditioned
hyperactivity; and (4) locomotor activity during exploration of a novel environment. Results: The two strains
showed consistent taste aversion at doses of amphetamine
that promoted opposite strain-dependent place conditioning. Both strains spent more time exploring the novel
rather than the known compartment of the place conditioning apparatus. Instead, only mice of the C57 strain
showed amphetamine-induced behavioral sensitization
and conditioned hyperactivity. However, temporary food
restriction did not affect strain differences for these
phenotypes. Finally, C57 mice were more active than
C. Orsini S. Puglisi-Allegra S. Cabib
Fondazione Santa Lucia, IRCCS, Rome, Italy
C. Orsini F. Buchini S. Puglisi-Allegra S. Cabib ())
Department of Psychology, University La Sapienza,
via dei Marsi 78, 00185 Roma , Italy
e-mail: simona.cabib@uniroma1.it
Tel.: +39-06-49917526
Fax: +39-06-49917712
P. V. Piazza
Laboratoire de Psychophysiologie des Comportements,
INSERM U588, Bordeaux, France

DBA in a novel environment and restricted feeding


abolished this strain-dependent difference. Conclusions: These results relate individual differences for
amphetamine-induced place conditioning with locomotor
response to amphetamine and novelty.
Keywords Addictive drugs Conditioning Context
discrimination Exploratory behavior Individual
predisposition to disease Inbred strain Life experience
Mice

Introduction
There is growing agreement on the main role of geneenvironment interaction in the etiology of psychiatric
disorders (Tsuang 2000). Recently, clinical (Vanyukov
and Tarter 2000) and preclinical (Cabib et al. 2000)
studies have pointed to the importance of exploring
genotypeenvironment interaction in the liability to drug
abuse. This type of research would take great advantage
from identification of behavioral phenotypes suitable for
murine modeling.
Amphetamine-induced place conditioning (CPP)
shows dramatic strain- and environment-dependent variability in mice. It has been demonstrated that mice of the
C57BL/6J (C57) inbred strain, show consistent preference
for an environment previously paired with amphetamine
over a large range of doses. Instead, mice of the DBA/2J
(DBA) inbred strain consistently avoid the amphetaminepaired environment. However, a brief experience of
restricted feeding promotes preference for an amphetamine-paired context in the avoiding DBA mice while not
affecting the response of C57 mice (Cabib et al. 2000),
therefore abolishing the strain differences.
A number of studies have characterized mice of the
C57 strain as alcohol and drug preferring, while DBA
mice appear to be resistant to drug and alcohol selfadministration (Carney et al. 1991; Seale and Carney
1991; Belknap et al. 1993; Meliska et al. 1994).
Moreover, food restriction is known to increase drug-self

265

administration in rodents (Carrol and Meisch 1984).


Finally, drug-induced CPP involves brain areas that are
activated by drug-associated stimuli in human addicts
(McBride et al. 1999; Sell et al. 2000; Hsu et al. 2002).
Therefore, the experience-dependent strain difference in
drug-induced place conditioning may represent a useful
model for the study of addictive mechanisms at a preclinical level. As a first step in the development of such
model, we evaluated behavioral phenotypes that might
share homologies with this response.
The processing of information related to context
identification and the sensitivity to the primary reinforcing properties of addictive substances are the phenotypes
that most likely play a role in individual susceptibility to
drug-induced place preference. Indeed, drug-induced CPP
is acquired through a Pavlovian conditioning process, by
which a neutral context repeatedly paired with the
unconditioned stimuli, acquires secondary reinforcing
properties. The positive incentive values of drugs of
abuse make the drug-paired but not the saline-paired
context able to elicit approach in the final discrimination
test. To the other end, drugs of abuse are known to
produce aversion (Cappell and LeBlanc 1977; Reicher
and Holman 1977; Goudie 1979), therefore the avoidance
of the amphetamine-paired context exhibited by mice of
the DBA strain may indicate higher sensitivity to the
negative reinforcing properties of addictive substances.
Moreover, mice of the C57 and DBA strains differ in their
performance in tests based on context discrimination (see
Rossi-Arnoud and Ammassari-Teule 1998 for review). In
particular, C57 mice are known to be good performers in
tasks based on spatial information, while DBA mice are
considered a model of a non-pathological deficit in tasks
based on spatial information, although their performance
is improved when provided with cues (Cabib et al. 2003).
Thus, in the first part of the present study, we
compared the two inbred strains for amphetamineinduced conditioned taste aversion (CTA), the standard
paradigm to evaluate aversive effects of addictive drugs
(Hunt and Amit 1987) and for discrimination of a novel
context in the same apparatus used in the CPP experiment. Significantly higher CTA scores in DBA mice were
considered to support the view of higher sensitivity to the
aversive effects of the psychostimulant in this strain of
mice. However, impaired discrimination of the novel
context in mice of the DBA strain would have indicated
that the testing condition was biased for spatial information.
In the second part of the study, we evaluated
behavioral sensitization and conditioned hyperactivity
induced by repeated administration of amphetamine.
Indeed, CPP training requires repeated administrations
in specific environments, and this procedure is known to
produce a progressive enhancement of the locomotor
effects of the drug (behavioral sensitization), and hyperactive response within the drug-paired environment in
drug-free state (conditioned hyperactivity, Stewart 1992;
Pierce and Kalivas 1997).

Since C57 and DBA mice show marked differences in


their susceptibility to develop behavioral sensitization and
conditioned hyperactivity (Puglisi-Allegra and Cabib
1997), we evaluated strain differences for both phenotypes following experience of the restricted feeding
protocol. The hypothesis to be tested was the elimination
of strain differences in previously food-restricted mice.
Finally, we considered levels of activity in a novel
environment since, although this phenotype correlates
with propensity to drug self-administration (Piazza et al.
1989), attempts at relating it to performance in CPP have
produced conflicting results (Erb and Parker 1994; Gong
et al. 1996; Kosten and Miserendino 1998). The homology between these two phenotypes was to be supported by
the finding of strain differences (C57>DBA) susceptible
to be abolished by the paradigm of restricted feeding that
eliminates strain differences for amphetamine-induced
CPP (Cabib et al. 2000).

Methods
Animals and housing
Male mice of the inbred C57BL/6JICo (C57) and DBA/2JICo
(DBA) strains (Charles River, Italy) were purchased at 6 weeks of
age and housed four to a cage on a 12-h/12-h light/dark cycle
(lights on 07001900 hours) for 3 weeks before behavioral testing.
All behavioral testing was performed during the second half of
the light phase in sound-attenuated cubicles, where temperature
was constant and a 30-W lamp was the only source of illumination.
Experiments were carried out in accordance with the Italian
Law (DL 116/92) on the use of animals in research.
Place conditioning
The CPP experiment was performed in boxes formed by two gray
Plexiglas chambers (151520 cm) and a central alley
(15520 cm). Two sliding doors (44 cm) connected the alley
to the chambers. In each chamber, two triangular parallelepipeds
(5520 cm) made of black Plexiglas and arranged to form
different patterns (always covering the same surface of the
chamber) were used as conditioned stimuli.
The training procedure was previously described in Cabib et al.
1996. Briefly, on day 1 (pretest), mice were free to explore the
entire apparatus for 15 min. During the following 8 days (conditioning phase), mice were confined daily for 40 min alternatively in
one of the two chambers. For each animal, during the conditioning
phase, one of the patterns was consistently paired with saline and
the other one with amphetamine. Pairings were balanced so that for
half of each experimental group amphetamine was paired with one
of the patterns and half of them with the other one. The first pairing
was with amphetamine for all subjects. Testing was conducted on
day 10 similarly to the pretest procedure. Test sessions were
videotaped and, later on, the time spent within each chamber of the
apparatus was quantified by an experimenter unaware of the
treatment conditions, using the Observer program (version 3.0,
System for Macintosh, Noldus, Wageningen 1997).
Taste conditioning
Training and testing for CTA were carried out in standard breeding
cages (262015 cm). The protocol chosen had previously
demonstrated a stringent parallelism between taste and place
aversion induced by naloxone in mice (Mucha and Walker 1987).

266
Drinking solutions were prepared daily in tap water and provided in
standard bottles equipped with a valve to prevent accidental
dropping/licking. The discriminative flavors were defined as salty
(12.5 mM monosodium glutamate + 128 mM NaCl) and acidic
(0.43 mM saccharine + 1.48 mM citric acid); these flavors are
known to be balanced in their preference (Mucha and Walker
1987). This CTA paradigm is based on a double choice in a no
deprived state.
Mice were first trained to drink in the test cage: after an
overnight water deprivation (about 15-hrs), animals were individually placed for thirty minutes into the test cage with a single bottle
filled with water and then returned to the home cage with free
access to water until the starting of the dark phase of the day/night
cycle. On the following day, the conditioning phase started and
mice received a total of six pairings in 2 days: three pairings of
amphetamine with one flavor and three pairings of saline with the
other. During the taste pairing session, each mouse was placed in a
test cage and a single bottle containing 60 ml flavored solution was
made available. Fifteen minutes later, mice were injected and then
returned to their home cages. Assignments of flavor/drug pairings
and order of pairings were counterbalanced. Ninety minutes after
the third pairing, mice had free access to water in their home cage
for 30 min. Pairing sessions were carried out between 0800 hours
and 1800 hours and interval time between sessions was not less
than 4 h and not longer than 14 h. Thus, every 4 h, the mice
received alternatively saline and amphetamine injections, for the
total of six trials. Ninety minutes after the last pairing, mice had
free access to water in their home cage until testing. Testing was
carried out the day after the conditioning phase, so those mice were
not tested in a water-deprived state. Mice were placed individually
into the test cage for 24 h. Each cage was provided with food and
two bottles containing the flavored solutions. Amount of liquid
consumption was measured.
Novelty discrimination
The apparatus was the same used for place conditioning. The
procedure consisted of training and test phase. On the training day,
mice were individually introduced in the central alley of the
apparatus and confined in the first compartment spontaneously
visited and left to explore it for 15 min. Twenty four hours later,
animals were once again introduced in the alley and left free to
explore the entire apparatus for a total of 20 min. Test sessions were
videotaped and, later on, time spent within each chamber of the
apparatus was quantified by an experimenter unaware of the
treatment conditions, using the Observer program (version 3.0,
System for Macintosh, Noldus, Wageningen 1997).
Restricted feeding procedure and locomotor activity
Food-restriction procedure consisted of single housing and a period
of partial food deprivation. The amount of food that was available
daily was progressively reduced and then adjusted to maintain the
loss of 20% of the initial body weight. After 12 days, food was
again available ad libitum. Testing started 2 days later. The
discrimination between the relative influence of individual housing
and restricted feeding on the observed effects was beyond the
purposes of this study. The main interest was to manipulate the
environment in the same way as reported in a study previously
conducted in this laboratory (Cabib et al. 2000).
Locomotor activity was measured in toggle floor boxes divided
into two compartments (2010 cm). Number of crossings between
the two compartments was recorded using a micro-switch connected to the tilting floor of the box.
Animals were tested daily for locomotor activity for four
consecutive days. On day 1, mice were first allowed to become
habituated to the test cage for 1 h. The number of crossings
recorded during this hour was used as locomotor response to
novelty. At the end of the habituation session, mice were placed
back in their home cages and left undisturbed for an additional

hour. Subsequently, mice were injected with amphetamine (1,


2 mg/kg) or saline and immediately placed into the test cage for 1 h.
This first test of the amphetamine-induced locomotor activity was
taken as a measure of the unconditioned effects of the drug. The
following 3 days, mice received an injection of the same dose of
amphetamine or saline and were immediately tested for locomotor
activity in the test cage, for a total of four challenges. On day 5, all
animals were injected with a saline challenge and tested as usual in
the test cage, and this measure was taken as conditioned hyperactivity.
Drug solutions
d-Amphetamine sulfate (Sigma) was dissolved in a vehicle solution
of 0.9% physiological saline, and injected intraperitoneally (i.p.) in
a volume of 10 ml/kg body weight.
Data analysis and statistics
For the first experiment, raw CPP and CTA data were transformed
into preference scores and used as dependent variables. Preference
scores were calculated as the following formulas: CPP = (amount of
time spent in the drug-paired compartment)/(amount of time spent
in both compartments)100; CTA = (consumed volume of the
drug-paired flavor/total fluid intake)100. Sixty mice (n=10 per
group) were used for amphetamine-induced place conditioning and
forty-eight mice (n=8 per group) were used for CTA. Experimental
groups consisted of free-fed C57 and DBA. Data from CPP and
CTA experiments were analyzed by a two-way ANOVA, factors
being: strains (C57 and DBA), and amphetamine dose (0, 1 and
2 mg/kg). Post-hoc analyses were carried out, when allowed, by
Duncans test.
In the second experiment time (seconds) spent in each
compartment of the CPP apparatus during the first 10 min of
testing was considered for either novelty discrimination or
amphetamine (2 mg/kg)-induced CPP. The decision of using the
first block of 10 min in both tests was based on the observation of a
time-dependent reduction of preference for the novel environment
in the second half of test in both strains. Data from the previous
experiment on amphetamine-induced CPP were used. Sixteen mice
(n=8 per group) were used for novelty discrimination. Data were
analyzed by two-way ANOVAs for repeated measures with strain
(C57 and DBA) as between factor and novelty (novel and known)
or amphetamine (Sal-paired and Amph-paired) as within factors.
Simple effects analyses within groups were performed when
allowed.
The third experiment involved 72 animals (n=6 per group).
Experimental groups consisted in free-fed (no prior food restriction
= F) C57 and DBA mice, and food-restricted (prior food restriction
= NF) C57 and DBA mice. Data from the behavioral sensitization
experiment were analyzed by ANOVA for repeated measures, with
time (days 1, 2, 3 and 4) as within factor, strain (C57 and DBA),
experimental condition (F and NF) and amphetamine dose (0, 1 and
2 mg/kg) as between factors. Data on locomotor activity during the
habituation phase were used for statistical analysis of locomotor
response to novelty. Forty-eight animals (n=12 per group) were
analyzed by a two-way ANOVA for repeated measures, with strain
(C57 and DBA) and experimental condition (F and NF) as between
subjects factors, time (six intervals of 10 min) as within subjects
factor.

Results
Dose-dependent effects of amphetamine on preference
scores in both CPP and CTA tests for C57 and DBA mice
are shown in Fig. 1. The ANOVA revealed only a
significant effect of amphetamine dose (F2,42=4.561,

267

Fig. 1 Preference scores (mean + SEM, see Methods for definitions) for amphetamine-paired compartment (place conditioning)
and taste (taste conditioning) in mice of the C57 and DBA strains.
*P<0.05 vs saline-treated mice (0) of the same strain (Duncans
test); P<0.05 vs mice of the other strain at the same dose
(Duncans test); #P<0.05 vs saline-treated mice (0) (Duncans test)

P<0.05) for taste conditioning. Comparisons between


means indicated significant amphetamine-induced avoidance for the drug-paired taste in both strains. For place
conditioning the ANOVA revealed a significant amphetamine dose  strain interaction (F2,54=8.075, P<0.001).
Post-hoc comparisons (Duncan test) did not reveal a
difference between strains when both compartments were
paired with saline. Furthermore, in this case, the preference scores were around 50, indicating that the conditioned cues used in this apparatus were not preferred or
avoided per se by mice of the two strains. By contrast,
significant differences between C57 and DBA mice were
observed when one of the compartments was paired with
either 1 mg or 2 mg amphetamine. Moreover, significant
preference for the drug-paired compartment was observed
in C57 mice, while a significant aversion was observed in
mice of the DBA strain (Duncan test).
Figure 2 shows the total time spent exploring the
amphetamine-paired compartment versus a saline-paired
one and a novel versus a known compartment by the two
strains. The ANOVA revealed a significant interaction
between the factors strain and amphetamine (F1,18=19.3,
P<0.0005) for CPP data and analyses of simple effects
indicated a significant preference for the amphetaminepaired compartment for C57 mice (F1,9=18.638, P<0.005)
and a significant avoidance of the amphetamine-paired
compartment for DBA mice (F1,9=7.218, P<0.05). By
contrast, the analysis of the novelty data showed only a
main effect of the factor novelty between compartment
(F1,14=18.773, P<0.001), indicating that both strains spent
more time within the novel compartment than the known
one.
Daily motor responses to amphetamine challenge (0, 1
and 2 mg/kg) in control (F) or prior food restricted (NF)
mice of the C57 and DBA strains are shown in Fig. 3. The
ANOVA revealed a significant interaction between the
factors time, strain and amphetamine dose (F6,180=5.57,
P<0.0001) and a significant interaction between time,
experimental condition and amphetamine dose
(F6,180=3.196, P<0.01). Post-hoc analysis after single
two-way ANOVAs for day 1 and day 4 revealed that on

Fig. 2 Time spent (seconds: mean + SEM) in the two compartments of the place conditioning apparatus in the 10-min test time
window for amphetamine-induced conditioning (left) and novelty
discrimination (right). *P<0.05 vs Sal-paired (simple effect)

Fig. 3 Daily locomotor response (mean crossings + SEM) to


different doses of amphetamine (0, 1, 2 mg/kg) by free-fed (F) or
previously food restricted (NF) mice of the C57 and DBA strains.
*P<0.05 vs Sal on the same day (Duncans test)

day 1, C57 mice showed a significant effect of the highest


dose of amphetamine in both experimental conditions (F
and NF), while the same dose increased the activity only
in food-deprived DBA. On day 4, a significant increase in
locomotor activity was observed after both doses of
amphetamine for free-fed C57, and for the previously
food-restricted C57 only at the highest dose. None of the
DBA groups showed an increased activity on day 4.
Finally, a significant increase of the locomotor response
to amphetamine between day 1 and day 4 (F1,5=27.36,
P<0.005) was observed in no prior food-restricted C57
mice challenged with the highest dose of amphetamine.
Figure 4 shows data on the conditioning effects of the
dose of 2 mg/kg of amphetamine as locomotor response to
a saline challenge in the test cage, after repeated
amphetamine or saline pairings. The statistical analysis
revealed significant main effects of strain (F1,40=37.65,
P<0.001), experimental condition (F1,40=6.956, P<0.05)
and amphetamine dose (F1,40=21.31, P<0.001). A significant interaction between the factors strain and amphetamine dose (F1,40=7.356, P<0.01) was also found. Posthoc analyses of data indicated that only mice of the C57
strain previously treated with 2 mg/kg amphetamine were
more active than the saline-pretreated mice regardless of
the experimental condition. Thus, only mice of the C57
strain showed amphetamine-induced conditioned hyper-

268

ed a significant difference between free-fed C57 and DBA


mice but no difference following food restriction.

Discussion

Fig. 4 Locomotor response to vehicle challenge (saline 10 ml/kg)


in free-fed (F) and previously food restricted (NF) mice of the C57
and DBA strains following repeated Saline (0) or amphetamine
(2 mg/kg, 2) pairings with the test cage. Data are expressed as total
crossings (mean + SEM). *P<0.05 vs 0 (Duncans test)

Fig. 5 Time course of locomotor activity, expressed as number of


crossings (mean + SEM), in a novel environment by free-fed (F)
and previously food restricted (NF) mice of the C57 and DBA
strains. In the top figure: total crossings (mean + SEM) over 1 h of
test. *P<0.05 vs C57 (Duncans test). P<0.05 vs the F group of the
same strain (Duncans test). #P<0.05 vs all others at the same time
point (Duncans test). * (top) P<0.005, strain difference (see
Results for details)

activity and this strain difference was unaffected by the


experience of food restriction.
Data on locomotor activity in a novel environment are
shown in Fig. 5. A two-way ANOVA for repeated
measures revealed a significant main effect of the factors
strain (F1,43=7.25, P<0.05), experimental condition
(F1,43=19.52, P<0.0001) and time (F5,215=77.68,
P<0.0001); and significant interactions time  strain
(F5,215=4.40, P<0.001), time  experimental condition
(F5,215=3.10, P<0.05), and global interaction between
strain, experimental condition and time (F5,215=2.839,
P<0.05). One-way analysis of total activity (60 min)
indicated a significant difference among groups
(F3,43=4.94, P<0.005). Post-hoc analyses (Fig. 5) indicat-

The present results suggest the possible homology


between sensitivity to the unconditioned locomotor
effects of amphetamine, the level of locomotor activation
in a novel environment, and amphetamine-induced place
conditioning. Indeed, locomotor response to amphetamine
and novelty were the only phenotypes, among those
investigated, that were susceptible to the gene-experience
interaction that influences the amphetamine-induced CPP.
The results obtained with our CTA paradigm do not
support the hypothesis of strain differences in sensitivity
to the aversive effects of amphetamine. Indeed, the
psychostimulant promoted taste aversion in both strains of
mice at the same doses inducing opposite strain-dependent place conditioning. It should be pointed out that the
data obtained in C57 mice are in agreement with the
observation that amphetamine is able to induce CTA
within the same dose range at which serves as positive
reinforcer (Reicher and Holman 1977).
CTA is a standard measure of the aversive effects of
addictive and non-addictive drugs, while CPP is used to
reveal positive incentive effects of addictive drugs (Hunt
and Amit 1987). Moreover, although the two paradigms
differ for a number of methodological aspects that do not
allow ruling out the possibilities of differences in
sensitivity to the aversive effects of amphetamine in the
two strains, the CTA protocol used had previously
demonstrated a stringent parallelism between taste and
place aversion induced by naloxone in mice (Mucha and
Walker 1987).
C57 and DBA mice show different performances in
discriminative tasks, based on the kind of informative
stimuli (spatial versus non-spatial) (Rossi-Arnoud and
Ammassari-Teule 1998; Cabib et al. 2003). The differences in the strategies used to learn about contexts might
have been involved in the strain-dependent effects of
amphetamine on place conditioning. To test this possibility, we performed a novelty discrimination task on the
two compartments of the CPP apparatus in drug-naive
animals. The results demonstrated that both strains of
mice are able to discriminate differences between the
contexts used for place conditioning and to reveal this
discrimination by preference.
Since there were no strain differences for either
amphetamine-induced CTA or for discrimination of a
novel compartment, we did not test these phenotypes for
the influence of the food restriction.
Strain differences for context-dependent sensitization
and conditioned hyperactivity induced by amphetamine
were unaffected by previous restricted feeding. It is worth
noting that food restriction enhanced the locomotor
response to amphetamine in DBA mice, as previously
described (Cabib et al. 2000), without affecting the
inability of the drug to promote behavioral sensitization in

269

this strain. These results add support to a lack of


relationship between behavioral sensitization and unconditioned locomotor effects of psychostimulant (see
Puglisi-Allegra and Cabib 1997 for review).
Finally, our data show strain differences that are
susceptible to be abolished by the experience of food
restriction for locomotor response to novelty. Indeed,
mice of the C57 strain showed higher levels of locomotor
activity in a novel environment than mice of the DBA
strain. Food restriction eliminated this strain difference by
enhancing the response in DBA strain more than in C57.
These data point to a strong relationship between
amphetamine-induced place conditioning and noveltyinduced locomotion. It should be pointed out that
previous studies have consistently failed to find a positive
correlation between levels of activity in a novel environment and the intensity of drug-induced CPP (Erb and
Parker 1994; Gong et al. 1996; Kosten and Miserendino
1998). The present data are not in conflict with these
reports. Indeed, we found a relationship between the
direction of drug-induced place conditioning (avoidance
versus preference) and the level of locomotion in a novel
environment.
The procedure used in these experiments does not
allow dissection of the relative contribution of individual
housing and restricted feeding to the observed effects.
Although individual housing does not affect strain
differences for the locomotor stimulating effects of
amphetamine (Cabib and Bonaventura 1997), it has major
strain-dependent influences on social behavior of male
mice (Puglisi-Allegra and Cabib 1985; Zocchi et al.
1994). However, these influences need at least 4 weeks of
individual housing to appear (Puglisi-Allegra and Cabib
1985; Zocchi et al. 1994). Moreover, repeated (2 h daily
for nine consecutive days, test on day 10) restraint stress
has been shown to eliminate strain differences for
amphetamine-induced locomotion in grouped mice (Badiani et al. 1992). Taken together, these considerations
suggest that the behavioral effects of our experimental
procedure are dependent on the stressful valence of the
experience of food restriction. Such conclusion is in line
with the observation that behavioral sensitization induced
by food restriction is dependent on corticosterone secretion (Deroche et al. 1995).
The selective effects of food restriction on strain
differences is not surprising. Indeed the experimental
procedure used in this study has been shown to reduce
(Cabib et al. 2000 and present results), promote (Cabib
and Bonaventura 1997) or enhance (Alcaro et al. 2000)
strain differences depending on the behavioral phenotypes
investigated. These considerations suggest that interactions between environmental and genetic variables are
extremely complex, and they support the idea that
manipulation of such variables may represent an effective
strategy for identifying shared homologies between
different phenotypes.
Thus, the only phenotypes that are susceptible to the
gene-experience interaction observed for amphetamineinduced CPP are the sensitivity to the unconditioned

locomotor effects of the psychostimulant (Cabib et al.


2000) and the level of locomotor activation in a novel
environment. These results suggest a possible homology
between these phenotypes and amphetamine-induced
place conditioning. In line with this hypothesis, noveltyand amphetamine-induced activity share with drug-induced CPP a common neural substrate, the mesolimbic
dopamine transmission (Wise and Bozarth 1987; Hooks et
al. 1991; Bardo et al. 1996; Piazza and LeMoal 1996;
McBride et al. 1999). Moreover, mesolimbic dopamine
transmission shows both strain differences and susceptibility to food restriction. Thus, C57 are characterized by
higher mesoaccumbens dopamine release in response to
amphetamine than DBA mice (Zocchi et al. 1998), and
recent results indicate that food restriction promotes
sensitization-like changes in the mesoaccumbens dopamine system of DBA mice (Cabib et al. 2002).
Acknowledgements This research was supported by Ministero
della Ricerca Scientifica e Tecnologica (COFIN: 20012002),
Ateneo 60% (20002001) and Facolt (2001).

References
Alcaro A, Cabib S, Ventura R, Puglisi-Allegra S (2002) Genotypeand experience-dependent susceptibility to depressive-like
responses in the forced-swimming test. Psychopharmacology
164:138143
Badiani A, Cabib S, Puglisi-Allegra S (1992) Chronic stress
induces strain-dependent sensitization to the behavioral effects
of amphetamine in the mouse. Pharmacol Biochem Behav
43:5360
Bardo MT, Donohew RL, Harrington NG (1996) Psychobiology of
novelty-seeking and drug-seeking behavior. Behav Brain Res
77:2343
Belknap JK, Crabbe JC, Young ER (1993) Voluntary consumption
of ethanol in 15 inbred mouse strains. Psychopharmacology
112:503510
Cabib S, Bonaventura N (1997) Parallel strain-dependent susceptibility to environmentally-induced stereotypes and stressinduced behavioral sensitization in mice. Physiol Behav
61:499506
Cabib S, Puglisi-Allegra S, Genua C, Simon H, Le Moal M, Piazza
PV (1996) Dose-dependent aversive and rewarding effects of
amphetamine as revealed by a new place conditioning apparatus. Psychopharmacology 125:9296
Cabib S, Orsini C, Le Moal M, Piazza PV (2000) Abolition and
reversal of strain differences in behavioral responses to drug of
abuse after a brief experience. Science 289:463465
Cabib S, VenturaR, Puglisi-Allegra S (2002) Opposite imbalances
between mesocortical and mesoaccumbens dopamine responses
to stress by the same genotype depending on living conditions.
Behav Brain Res 129:179185
Cabib S, Pascucci T, Ventura R, Romano V, Puglisi-Allegra S
(2003) The behavioral profile of severe mental retardation in a
genetic mouse model of phenylketonuria. Behav Genetics
33:301310
Cappell H, LeBlanc AE (1977) Parametric investigations of the
effects of prior exposure to amphetamine and morphine on
conditioned gustatory aversion. Psychopharmacology 51:265
271
Carney JM, Landrum RW, Cheng MS, Seale TW (1991) Establishment of chronic intravenous drug self-administration in the
C57BL/6 mouse. Neuroreport 2:477480
Carrol ME, Meisch RA (1984) Increased drug-reinforced behavior
due to food deprivation. Adv Behav Pharmacol 4:4788

270
Deroche V, Marinelli M, Maccari S, Le Moal M, Simon H, Piazza
PV (1995) Stress-induced sensitization and glucocorticoids. I.
Sensitization of dopamine-dependent locomotor effects of
amphetamine and morphine depends on stress-induced corticosterone secretion. J Neurosci 15:781758
Erb SM, Parker LA (1994) Individual differences in noveltyinduced activity do not predict strength of amphetamineinduced place conditioning. Pharmacol Biochem Behav
48:581586
Gong W, Neill DB, Justice JB Jr (1996) Locomotor response to
novelty does not predict cocaine place preference conditioning
in rats. Pharmacol Biochem Behav 53:191196
Goudie AJ (1979) Aversive stimulus properties of drugs. Neuropharmacology 18:971979
Hooks MS, Jones GH, Smith AD, Neill DB, Justice JB (1991)
Response to novelty predicts the locomotor and nucleus
accumbens dopamine response to cocaine. Synapse 91:121128
Hunt T, Amit Z (1987) Conditioned taste aversion induced by selfadministered drugs: paradox revisited. Neurosci Behav Rev
11:107130
Hsu EH, Schroeder JP, Packard MG (2002) The amygdala mediates
memory consolidation for an amphetamine-conditioned place
preference. Behav Brain Res 129:93100
Kosten TA, Miserendino MJ (1998) Dissociation of novelty- and
cocaine-conditioned locomotor activity from cocaine place
conditioning. Pharmacol Biochem Behav 60:785791
McBride WJ, Murphy JM, Ikemoto S (1999) Localization of brain
reinforcement mechanism: intracranial self-administration and
intracranial place-conditioning studies. Behav Brain Res
101:129152
Meliska CJ, Bartke A, McGlacken G, Jensen RA (1994) Ethanol,
nicotine, amphetamine consumption and preferences in C57BL/
6 and DBA/2 mice. Pharmacol Biochem Behav 50:619626
Mucha RF, Walker MJK (1987) Aversive property of opioid
receptor blockade in drug-naive mice. Psychopharmacolgoy
93:483488
Piazza PV, Le Moal M (1996) Pathophysiological basis of
vulnerability to drug abuse: role of an interaction between
stress, glucocorticoids, and dopaminergic neurons. Annu Rev
Pharmacol Toxicol 36:359378
Piazza PV, Deminiere JM, Le Moal M, Simon H (1989) Factors
that predict individual vulnerability to amphetamine selfadministration. Science. 245:15111513

Pierce RC, Kalivas PW (1997) A circuitry model of the expression


of behavioral sensitization to amphetamine-like psychostimulants. Brain Res Rev 25:192216
Puglisi-Allegra S, Cabib S (1985) The effect of age on two kinds of
aggressive behavior in inbred strains of mice. Dev Psychobiol
19:477482
Puglisi-Allegra S, Cabib S (1997) Psychopharmacology of dopamine: the contribution of comparative studies in inbred mice.
Prog Neurobiol 51:637661
Reicher M, Holman EW (1977) Location preference and flavour
aversion reinforced by amphetamine in rats. Anim Learn Behav
5:343346
Rossi-Arnoud C, Ammassari-Teule M (1998) What do comparative
studies of inbred mice add to current investigations on the
neural basis of spatial behaviors? Exp Brain Res 123:3644
Seale TW, Carney JM (1991) Genetic determinants of susceptibility
to the rewarding and other behavioral actions of cocaine. J
Addict Dis 10:141162
Sell LA, Morris JS, Bearn J, Frackowiak RSJ, Friston KJ, Dolan RJ
(2000) Neural responses associated with cue evoked emotional
states and heroin in opiate addicts. Drug Alcohol Depend
60:207216
Stewart J (1992) Conditioned stimulus control of the expression of
sensitization of the behavioral activating effects of opiate and
stimulant drugs. In: Gormezano I, Wasserman EA (eds)
Learning and memory: behavioral and biological substrates.
Erlbaum, Hillsdale
Tsuang M (2000) Schizophrenia: genes and environment. Biol
Psychiatry 47:210220
Vanyukov MM, Tarter RE (2000) Genetic studies of substance
abuse. Drug Alcohol Depend 59:101123
Wise RA, Bozarth MA (1987) A psychomotor stimulant theory of
addiction. Psychol Rev 94:469492
Zocchi A, Cabib S, Puglisi-Allegra S (1994) Opposite straindependent differences for intermale aggressive behavior elicited by individual housing and housing with a female in the
mouse. Aggress Behav 20:305314
Zocchi A, Orsini C, Cabib S, Puglisi-Allegra S (1998) Parallel
strain-dependent effect of amphetamine on locomotor activity
and dopamine release in the nucleus accumbens: an in vivo
study in mice. Neuroscience 82:521528