Conditioned place aversion induced by intragastric administration of ethanol in rats

Most experiments investigating ethanol-induced place conditioning in rats have produced conditioned place aversion (CPA). In one of the few reports of ethanol-induced conditioned place preference (CPP) in rats, selectively bred alcohol-preferring (msP) rats showed CPP in a biased procedure when ethanol was administered via intragastric (IG) catheter but not when ethanol was administered via intraperitoneal injection or by gavage. This finding suggests the importance of both route of administration and genetic variables to the outcome of place conditioning studies. We conducted three experiments examining place conditioning induced by IG ethanol in genetically heterogeneous rats to test the generality of the earlier finding. We employed an unbiased procedure that is more sensitive to detecting preference changes in either direction (preference or aversion). Ethanol-naive (Experiment 1) and ethanol-experienced Sprague-Dawley rats (Experiment 2) showed robust CPA. In Experiment 3, infusion rate was varied to see if the CPA observed in Experiments 1 and 2 was a result of the rapidity of the transition from the sober to the intoxicated states. Both groups showed strong CPA. Overall, the present findings are consistent with previous findings of CPA in heterogeneous rats, suggesting that the aversive postabsorptive effects of ethanol produce CPA.     

Flavor preferences conditioned by intragastric ethanol with limited access training

In a prior study, ad libitum fed rats learned a strong preference (90%) for a flavored saccharin solution (conditioned stimulus, CS+) paired with concurrent intragastric (IG) infusions of 5% ethanol over another flavor (CS-) paired with water infusions in unlimited access sessions (22 h/day). The present study expanded the investigation of ethanol-conditioned preferences to limited access sessions (30 min/day). Experiment 1 revealed that ad lib or food-restricted rats failed to develop a CS+ preference using the same CS solutions (0.05% Kool-Aid+0.2% saccharin) and IG infusions that were effective with long-term training. Experiments 2 and 3 mimicked the parameters from a report of successful ethanol conditioning in deprived rats: ethanol (0.5 g/kg) or water was infused intragastrically 5 min before access to sweetened CS solutions flavored with HCl or NaCl. Rats learned to prefer the ethanol-paired CS+ when the flavors were mixed with 5% sucrose but not when mixed with 0.2% saccharin. Experiment 4 revealed that 5% sucrose solutions flavored with 0.25% Kool Aid also supported flavor preference conditioning by IG ethanol (0.5 g/kg). CS+ preferences were obtained in rats trained with ethanol infused 5 min before or concurrent with CS+ intake, but not in rats trained with ethanol infused 30 min before CS+ intake. These data confirm that flavor preferences can be conditioned by IG ethanol using a limited access procedure. However, in contrast to 22 h/day training, 30 min/day training requires more intense CS flavors and a nutritive sweetener. The preference reinforcing actions of ethanol may develop slowly and are thus most effective with long training sessions or when intense CS flavors are used in short training sessions.     

FOS expression induced by an ethanol-paired conditioned stimulus

To identify brain areas involved in ethanol-induced Pavlovian conditioning, brains of male DBA/2J mice were immunohistochemically analyzed for FOS expression after exposure to a conditioned stimulus (CS) previously paired with ethanol (2 g/kg) in two experiments. Mice were trained with a procedure that normally produces place preference (Before: ethanol before the CS) or one that normally produces place aversion (After: ethanol after the CS). Control groups received unpaired ethanol injections in the home cage (Delay) or saline only (Na?ve). On the test day, mice were exposed to the 5-min CS 90 min before sacrifice. Before groups showed a conditioned increase in activity, whereas the After group showed a conditioned decrease in activity. FOS expression after a drug-free CS exposure was significantly higher in Before-group mice than in control mice in the bed nucleus of the stria terminalis (Experiment 1) and anterior ventral tegmental area (Experiments 1-2). Conditioned FOS responses were also seen in areas of the extended amygdala and hippocampus (Experiment 2). However, no conditioned FOS changes were seen in any brain area examined in After-group mice. Overall, these data suggest an important role for the mesolimbic dopamine pathway, extended amygdala and hippocampus in ethanol-induced conditioning.     

The effects of pimozide on the establishment of conditioned reinforcement as a function of the amount of conditioning

In an attempt to understand some inconsistent findings, the present experiment investigated the effects of pimozide, a dopamine (DA) receptor blocker, on the establishment of conditioned reinforcement as a function of the amount of conditioning. In Experiment 1, rats received three phases of training in a two-lever box. The pre-exposure phase measured the operant rates of pressing the levers; one produced a 3-s tone and the other turned the lights off for 3 s. In the conditioning phase, with the levers absent, the light-off stimulus was paired with food for two or four sessions. The test phase again measured the rate of pressing the levers. Conditioned reinforcement was shown by a relative increase in responding on the light lever during the test. Of the groups receiving four conditioning sessions, pimozide (0.5, 1.0, 2.0 and 4.0 mg/kg) produced a dose-dependent attenuation of conditioned reinforcement, those rats treated with 4.0 mg/kg failing to demonstrate a significant effect. When 2 conditioning days were employed, pimozide treatment also produced a dose-dependent attenuation; however, in these less conditioned animals 2.0 mg/kg blocked the effect. The possibility that pimozide produced a conditioned taste aversion to the food was ruled out in Experiment 2. These data suggest that DA transmission may be necessary for the establishment of conditioned reinforcement and that the effects of receptor blockade may be related to the amount of conditioning.     

Morphine conditioned place preference in the hamster

Two experiments investigated the ability of morphine to produce a conditioned place preference in the hamster. In Experiment 1, it was found that a 15 mg/kg dose of morphine produced a conditioned place preference after eight conditioning trials. In addition, naloxone (0.4 mg/kg blocked the development of the morphine-conditioned place preference and itself produced a conditioned place aversion after four conditioning trials. In Experiment 2, the effects of four doses of morphine (0, 2.5, 5 and 15 mg/kg) on the acquisition of a conditioned place preference were studied. Only the 15 mg/kg dose produced a significant place preference. Compared to similar findings in the rat, the present results indicate that a relatively high dose of morphine is required to produce a conditioned place preference in the hamster.     

Classically conditioned motor effects do not occur with cocaine in an unbiased conditioned place preferences procedure

Classical conditioning and behavioural sensitisation of motor activity induced with cocaine (10mg/kg, i.p.) were examined using an unbiased two-compartment conditioned place preference (CPP) procedure. Habituation of the rats to the testing environment prior to training was varied (i.e. either the rats were habituated to the environment for three 30min sessions or they were not) in order to examine a possible influence of latent inhibition on conditioned locomotion or behavioural sensitisation. Furthermore, rats were either trained with an explicit CS+ (cocaine-paired compartment) and CS- (vehicle-paired compartment), or else they were trained with no barrier between the compartments (effectively a single-compartment procedure with no explicit CS-) in order to examine a possible influence of stimulus change (training rats while confined to one compartment, but testing with no barrier between compartments). On a drug-free test day with free access to both compartments, rats previously exposed to cocaine in one compartment (CS+) and vehicle in the second compartment (CS-) spent more time in the CS+ compartment (conditioned place preference). However, under no circumstance was the rate of motor activity higher in the CS+ compartment than in the CS- compartment, as would be expected if cocaine-induced motor activity was classically conditioned to contextual cues. Whether or not increased activity extinguished with repeated drug-free exposures to previously drug-paired contexts depended on habituation experience. In addition, both habituation and current access to compartments (free or restricted) determined the presence of post-extinction sensitisation to a challenge dose of cocaine (7.5mg/kg). Classical conditioning and non-associative sensitisation, independently or together, cannot account for this pattern of results.     

Reducing endocannabinoid metabolism with the fatty acid amide hydrolase inhibitor, URB597, fails to modify reinstatement of morphine-induced conditioned floor preference and naloxone-precipitated morphine withdrawal-induced conditioned floor avoidance

The potential of the fatty acid amide hydrolase (FAAH) inhibitor, URB597, to modify drug prime-induced reinstatement of morphine-induced conditioned floor preference or naloxone-precipitated morphine withdrawal-induced conditioned floor avoidance was evaluated. In Experiment 1, morphine-induced conditioned floor preference was established across 4 conditioning trials. Following extinction training (4 trials), rats were pretreated with URB597 or vehicle prior to a morphine prime or a saline prime. Morphine reinstated the previously extinguished floor preference, but URB597 did not modify the strength of the reinstated preference. In Experiment 2, naloxone-precipitated morphine withdrawal-induced conditioned floor avoidance was established across 2 conditioning trials. Following extinction training (14 trials), rats were pretreated with URB597 or vehicle prior to a saline prime or a morphine withdrawal prime. The morphine withdrawal prime reinstated the previously extinguished floor avoidance, but URB597 did not modify the strength of reinstated avoidance. These results suggest that under the conditions in which URB597 promotes extinction (e.g., Manwell et al. (2009)) it does not interfere with drug-induced reinstatement of either conditioned floor preference or avoidance. That is, although activation of the endocannabinoid (eCB) system promotes extinction of aversive learning, it may not prevent reinstatement of that aversion by re-exposure to the aversive treatment.     

Failure to establish a conditioned place preference with ethanol in rats

Previous studies have demonstrated that many drugs of abuse are able to produce a conditioned place preference in rats. We sought to determine if ethanol, injected in a wide range of doses, could also produce a conditioned place preference. Statistical analysis of our results indicated that the IP administration of the drug (50, 100, 150, 300, 600, 800, or 1000 mg/kg) failed to produce either a conditioned place preference or aversion compared to vehicle injected control rats. Under similar testing conditions a conditioned place preference was obtained with amphetamine (2 mg/kg) and this preference was not secondary to conditioned hyperactivity. In another experiment, rats were injected with ethanol through indwelling jugular cannulae at doses similar to those reported [24,26] to support (1, 2 mg/kg) or not to support (8 mg/kg) self-administration by rats. We also failed to obtain a conditioned place preference using these doses. Blood and brain ethanol levels, determined 1, 2 or 5 minutes after the administration of 2 mg/kg (IV) indicated very low ethanol levels. These results may suggest that rats do not self-administer ethanol for its intoxicating properties, and that the affective state produced by ethanol administration per se is not readily conditionable to environmental cues.     

Conditioned activation induced by ethanol: role in sensitization and conditioned place preference.

Previous studies of ethanol-induced activation and place preference conditioning have shown that repeated exposure to ethanol produces sensitization to ethanol's locomotor activating effect in mice. This experiment was designed to determine whether the behavioral sensitization to ethanol that occurs during place preference conditioning is due to development of a Pavlovian conditioned activity response. Mice (DBA/2J) in the experimental group (BEFORE) received four pairings of a distinctive floor stimulus with ethanol (2 g/kg, IP); a different floor stimulus was paired with saline (counterbalanced). Mice in two control groups were exposed equally to each floor stimulus and were handled and injected as often as experimental mice. One control group (AFTER) always received ethanol in the home cage 1 h after exposure to the floor stimulus, while the other control group (NO-DRUG) never received ethanol during conditioning. BEFORE group mice showed a significant conditioned place preference, whereas control mice did not. Activity tests after saline or ethanol indicated higher activity levels in BEFORE mice compared to control mice, regardless of floor stimulus. Moreover, BEFORE mice were more active on their CS+ floor than on their CS- floor during saline tests; activity was equally elevated on both floors during ethanol tests. These results support the hypothesis that sensitization to ethanol's activating effect is mediated by Pavlovian conditioning. Further, they suggest that place conditioning established-associative control by two kinds of stimuli; the specific tactile cues serving as CS+ and CS- and the general environmental cues common to both CS+ and CS- trials.(ABSTRACT TRUNCATED AT 250 WORDS)     

Ethanol-induced conditioned place aversion in mice

Previous studies have shown that ethanol produces conditioned place preference (CPP) in mice when injections are given immediately before exposure to the conditioned stimulus (CS). Paradoxically, however, injection of ethanol immediately after the CS produces conditioned place aversion (CPA). Four experiments were conducted to characterize the parametric boundaries of CPA produced by post-CS ethanol exposure. Experiment 1 showed that CPA is positively related to ethanol dose, with significant CPA at 2 and 4 g / kg, but not at 1 g / kg. Experiment 2 revealed an inverse relationship between CPA and trial duration, i.e. significant CPA occurred when the trial duration was 5, 15 or 30 min, but not when it was 60 or 90 min. Experiment 3 indicated that ethanol pre-exposure (eight daily injections) significantly reduced subsequent development of CPA. Finally, experiment 4 showed that repeated exposure to the CS alone (six 30 min exposures to each CS) after CS-ethanol pairings produced complete extinction of CPA. The same extinction procedure also completely eliminated CPP induced by pre-CS injections of ethanol. Overall, these studies demonstrate that CPA induced by post-CS ethanol injection is influenced by many of the same variables that affect CPP produced by pre-CS ethanol injection in mice. However, these findings do not resolve the issue of whether the 'before-versus-after' effect in ethanol place conditioning is better explained by assuming ethanol produces only rewarding effects or by assuming that ethanol produces both rewarding and aversive effects.     

Infant rats exhibit aversive learning mediated by ethanol's orosensory effects but are positively reinforced by ethanol's post-ingestive effects

Previous work suggest aversive and appetitive hedonic effects of intraorally delivered EtOH in pre-weanling rats. Pups are reluctant to perform an operant response when reinforced with intraoral EtOH infusions, a result suggesting aversive orosensory properties of EtOH. Yet, post-absorptive effects of ethanol seem capable of supporting appetitive conditioning. Two experiments were conducted to test this phenomenon. Both included a pre-exposure phase (postnatal day 13, PD13) comprising intraoral stimulation with water or EtOH. In Experiment 1, pups were given pairings between a tactile conditioned stimulus (CS) and intraoral infusions of EtOH or water. A subsequent tactile preference test revealed that pups spent significantly less on the EtOH-related CS relative to time spent on the alternative CS. In Experiment 2 pups were exposed to a texture CS (sandpaper) while intraorally infused with EtOH or during a later EtOH post-infusion interval. A tactile locational test conducted on PD16 indicated that EtOH-pre-exposed animals that experienced sandpaper paired with EtOH's post-absorptive effects exhibited a significant preference for the CS, even relative to a control group that experienced non-reinforced exposure to the tactile CS during conditioning. These results confirm that intraoral ethanol acts as an aversive tastant. A brief pre-exposure to EtOH allows later expression of appetitive learning mediated by the drug's post-ingestive effects.     

Aversion instead of preference learning indicated by nicotine place conditioning in rats

Although nicotine is a drug of abuse for millions of smokers, it has been difficult to demonstrate clearly the motivational properties of nicotine with rats using the conditioned place preference (CPP) paradigm. The first experiment attempted to replicate CPPs reported by other researchers using nicotine doses of 0.4, 0.8, and 1.2 mg/kg. There was a trend for all three doses to produce aversions, but it was significant only for the 0.8 mg/kg dose. Exposures to the CS alone extinguished aversions, but a priming dose (0.2 mg/kg) of nicotine given after extinction produced aversions only in animals exposed to 1.2 mg/kg. Experiment 2 tested whether preexposure to morphine or nicotine would sensitize animals to nicotine's reinforcing effects. In this experiment, rats were exposed to either six nicotine (0.6 mg/kg) or morphine (1.0 mg/kg) dosings prior to preference conditioning. Neither preferences nor aversions were observed in any group following subsequent conditioning with 0.6 mg/kg nicotine. The results suggest that previous observations of preference effects may have been due to specific procedural factors or may have depended on negative reinforcement due to stress reduction.     

Apparatus bias and place conditioning with ethanol in mice

RATIONALE: Although the distinction between "biased" and "unbiased" is generally recognized as an important methodological issue in place conditioning, previous studies have not adequately addressed the distinction between a biased/unbiased apparatus and a biased/unbiased stimulus assignment procedure. Moreover, a review of the recent literature indicates that many reports (70% of 76 papers published in 2001) fail to provide adequate information about apparatus bias. This issue is important because the mechanisms underlying a drug's effect in the place-conditioning procedure may differ depending on whether the apparatus is biased or unbiased. OBJECTIVES: The present studies were designed to assess the impact of apparatus bias and stimulus assignment procedure on ethanol-induced place conditioning in mice (DBA/2 J). A secondary goal was to compare various dependent variables commonly used to index conditioned place preference. METHODS: Apparatus bias was manipulated by varying the combination of tactile (floor) cues available during preference tests. Experiment 1 used an unbiased apparatus in which the stimulus alternatives were equally preferred during a pre-test as indicated by the group average. Experiment 2 used a biased apparatus in which one of the stimuli was strongly preferred by most mice (mean % time on cue = 67%) during the pre-test. In both studies, the stimulus paired with drug (CS+) was assigned randomly (i.e., an "unbiased" stimulus assignment procedure). Experimental mice received four pairings of CS+ with ethanol (2 g/kg, i.p.) and four pairings of the alternative stimulus (CS-) with saline; control mice received saline on both types of trial. Each experiment concluded with a 60-min choice test. RESULTS: With the unbiased apparatus (experiment 1), significant place conditioning was obtained regardless of whether drug was paired with the subject's initially preferred or non-preferred stimulus. However, with the biased apparatus (experiment 2), place conditioning was apparent only when ethanol was paired with the initially non-preferred cue, and not when it was paired with the initially preferred cue. These conclusions held regardless of which dependent variable was used to index place conditioning, but only if the counterbalancing factor was included in statistical analyses. CONCLUSIONS:These studies indicate that apparatus bias plays a major role in determining whether biased assignment of an ethanol-paired stimulus affects ability to demonstrate conditioned place preference. Ethanol's ability to produce conditioned place preference in an unbiased apparatus, regardless of the direction of the initial cue bias, supports previous studies that interpret such findings as evidence of a primary rewarding drug effect. Moreover, these studies suggest that the asymmetrical outcome observed in the biased apparatus is most likely due to a measurement problem (e.g., ceiling effect) rather than to an interaction between the drug's effect and an unconditioned motivational response (e.g., "anxiety") to the initially non-preferred stimulus. More generally, these findings illustrate the importance of providing clear information on apparatus bias in all place-conditioning studies.     

Localization of genes influencing ethanol-induced conditioned place preference and locomotor activity in BXD recombinant inbred mice

Genetic differences in ethanol's ability to induce conditioned place preference were studied in 20 BXD Recombinant Inbred (RI) mouse strains and in the C57BL/6J and DBA/2J progenitor strains. Male mice from each strain were exposed to a Pavlovian conditioning procedure in which a distinctive floor stimulus (CS+) was paired four times with ethanol (2 g/kg). A different floor stimulus (CS-) was paired with saline. Control mice were injected only with saline. Floor preference testing without ethanol revealed significant genetic differences in conditioned place preference, with some strains spending nearly 80% time on the ethanolpaired floor while others spent only 50% (i.e., no preference). Control mice showed genetic differences in unconditioned preference for the floor cues, but unconditioned preference was not genetically correlated with conditioned preference. There were also substantial genetic differences in ethanol-stimulated activity, but contrary to psychomotor stimulant theory, ethanol-induced activity on conditioning trials was not positively correlated with strength of conditioned place preference. However, there was a significant negative genetic correlation (r=–0.42) between test session activity and preference. Quantitative trait loci (QTL) analyses showed strong associations (P<0.01) between conditioned place preference and marker loci on chromosomes 4, 8, 9, 18 and 19. Weaker associations (0.01<P<0.05) were identified on several other chromosomes. Analysis also yielded several significant QTL for unconditioned preference, ethanol-stimulated activity, and sensitization. Overall, these data support the conclusion that genotype influences ethanol-induced conditioned place preference, presumably via genetic differences in sensitivity to ethanol's rewarding effects. Moreover, several chromosomal regions containing candidate genes of potential relevance to ethanol-induced conditioned place preference have been identified.     

Agmatine attenuates methamphetamine-induced conditioned place preference in rats

The polyamine agmatine modulates a variety of behavioral effects including the abuse-related effects of opioids and has been proposed as a potential medication candidate for the treatment of opioid abuse. However, little is known of the effects of agmatine on the abuse-related effects of other drugs of abuse. This study examined the effects of agmatine on the rewarding effects of methamphetamine in rats using a conditioned place preference paradigm. Methamphetamine (0.1-1.0mg/kg) dose-dependently increased the time spent in methamphetamine-paired side (place preference). Agmatine, at doses that did not produce place preference or aversion (10-32mg/kg), significantly decreased the development of methamphetamine-induced place preference when agmatine was administered in combination with methamphetamine during place conditioning. Agmatine also significantly decreased the expression of methamphetamine-induced place preference when an acute injection of agmatine was given immediately before test session. These doses of agmatine do not alter the motor activity in rats, suggesting that the observed attenuation of methamphetamine-induced place preference was not due to general behavioral disruption. Together, these data suggests that agmatine attenuates the rewarding effects of methamphetamine and may be able to modulate the abuse liability of methamphetamine.     

Dissociation of Novelty- and Cocaine-Conditioned Locomotor Activity From Cocaine Place Conditioning

High locomotor response to novelty is associated with ease of drug self-administration but does not predict greater place-conditioning effects of drugs. Yet, the latter reflects context conditioning and high responders (HR), compared to low responders (LR), show greater conditioned locomotor effects. Conditioned locomotor effects may occur in place conditioning, perhaps confounding its measure. To examine whether conditioned locomotor effects occur in place conditioning, the present study classified rats as HR vs. LR by using approximately the two extreme 15% percentiles of the distributions. The place conditioning and locomotor sensitizing effects of cocaine were tested. In Experiment 1, HR rats exhibited more crossings between compartments but did not differ from LR rats in cocaine place conditioning. Further, both groups showed increased crossings at test compared to baseline, indicative of a conditioned locomotor effect. In Experiment 2, HR rats showed greater acute locomotor activation to cocaine, whereas LR rats tend to show greater locomotor sensitization. Finally, in Experiment 3, HR rats showed habituation in locomotor responses, whereas LR rats did not. Results of these studies suggest that inherent and conditioned locomotor activity levels are dissociated from place-conditioning effects.     

dextro-Morphine attenuates the morphine-produced conditioned place preference via the sigma(1) receptor activation in the rat

An unbiased conditioned place preference paradigm was used to evaluate the effect of dextro-morphine on the morphine-produced reward in male CD rats. Morphine sulfate (1-10 mg/kg) given intraperitoneally dose-dependently produced the conditioned place preference. Pretreatment with dextro-morphine at a dose from 0.1 to 3 μg/kg given subcutaneously dose-dependently attenuated the morphine-produced conditioned place preference. However, dextro-morphine at a higher dose 100 μg/kg did not affect the morphine-produced conditioned place preference. Thus, dextro-morphine pretreatment induces a U-shaped dose-response curve for attenuating the morphine-produced conditioned place preference. The attenuation of the morphine-produced conditioned place preference was reversed by the pretreatment with the sigma₁ receptor antagonist BD1047 (N-[2-(3,4-Dichlorophenyl)ethyl]-N-methyl-2-(dimethylamino)ethylamine dihydrobromide. dextro-Morphine or BD1047 given alone did not affect the baseline place conditioning. It is concluded that dextro-morphine attenuated the morphine-produced conditioned place preference via the sigma₁ receptor activation.     

Selective D1 and D2 dopamine agonists produce opposing effects in place conditioning but not in conditioned taste aversion learning

The neurotransmitter, dopamine (DA), has been implicated in place conditioning but the role of D1 and D2 receptors has not been investigated. In Experiment 1, the effects of SKF 38393 (0, 0.01, 0.1, 1.0, 10.0 mg/kg) and quinpirole (0, 0.01, 0.1, 1.0, 2.0, 4.0 mg/kg), preferential D1 and D2 receptor agonists, respectively, were evaluated and compared to (+)-amphetamine (0, 0.01, 0.1, 1.0, 2.0, 4.0 mg/kg). The experiment consisted of three phases. During the preexposure phase, rats explored two distinctive end compartments adjoined by a small tunnel. The time spent in each compartment was recorded. During the 8-day conditioning phase, rats were treated with drug and confined to one compartment for 30 min. On alternate days, rats received saline and were placed in the opposite compartment. Test days occurred over the remaining three days during which drug-free animals explored both compartments. Rats conditioned with (+)-amphetamine demonstrated a dose-dependent increase in time spent in the drug-paired environment from preexposure to test indicating the establishment of a conditioned place preference. Treatment with quinpirole also resulted in a conditioned place preference, however, only an intermediate dose was effective. In contrast, SKF 38393 produced a dose-dependent decrease in time spent on the drug-paired side suggesting the establishment of a place aversion. The idea that D1 receptors may be exclusively involved in mediating the aversive properties of psychomotor stimulants was tested in Experiment 2 employing a conditioned taste aversion paradigm. The results did not support this notion; it was found that both quinpirole and SKF 38393 produced a conditioned taste aversion.(ABSTRACT TRUNCATED AT 250 WORDS)     

Fenamate-induced enhancement of heterologously expressed HERG currents in Xenopus oocytes

In the present study, the effects of intra-nucleus accumbens injection of L-arginine, a nitric oxide (NO) precursor, and N(G)-nitro-L-arginine methyl ester (L-NAME), a nitric oxide synthase (NOS) inhibitor, on morphine-induced conditioned place preference in male Wistar rats were investigated. Our data showed that subcutaneous (s.c.) injection of morphine sulphate (0.5-10 mg/kg) significantly increased the time spent in the drug-paired compartment in a dose-dependent manner. Intra-accumbens administration of L-arginine (0.03 and 0.05 microg/rat) with an ineffective dose of morphine (0.5 mg/kg) elicited significant conditioned place preference, while intra-accumbens administration of L-NAME (0.3, 0.1 and 1 microg/rat) decreased the acquisition of conditioned place preference induced by morphine (7.5 mg/kg). The response to different doses of L-arginine was decreased by L-NAME (0.03 microg/rat). L-Arginine and L-NAME by themselves did not elicit any effect on place conditioning. Intra-accumbens administration of L-arginine but not L-NAME significantly decreased the expression of morphine (7.5 mg/kg)-induced place preference. The attenuation of already established morphine-induced place preference on the test day by L-arginine was inhibited by L-NAME. The results indicate that NO may be involved in the acquisition and expression of morphine-induced place preference.     

The mGluR5 Antagonist MTEP Dissociates the Acquisition of Predictive and Incentive Motivational Properties of Reward-Paired Stimuli in Mice

An environmental stimulus paired with reward (a conditioned stimulus; CS) can acquire predictive properties that signal reward availability and may also acquire incentive motivational properties that enable the CS to influence appetitive behaviors. The neural mechanisms involved in the acquisition and expression of these CS properties are not fully understood. The metabotropic glutamate receptor, mGluR5, contributes to synaptic plasticity underlying learning and memory processes. We examined the role of mGluR5 in the acquisition and expression of learning that enables a CS to predict reward (goal-tracking) and acquire incentive properties (conditioned reinforcement). Mice were injected with vehicle or the mGluR5 antagonist, MTEP (3 or 10 mg/kg), before each Pavlovian conditioning session in which a stimulus (CS+) was paired with food delivery. Subsequently, in the absence of the primary food reward, we determined whether the CS+ could reinforce a novel instrumental response (conditioned reinforcement) and direct behavior toward the place of reward delivery (goal-tracking). MTEP did not affect performance during the conditioning phase, or the ability of the CS+ to elicit a goal-tracking response. In contrast, 10 mg/kg MTEP given before each conditioning session prevented the subsequent expression of conditioned reinforcement. This dose of MTEP did not affect conditioned reinforcement when administered before the test, in mice that had received vehicle before conditioning sessions. Thus, mGluR5 has a critical role in the acquisition of incentive properties by a CS, but is not required for the expression of incentive learning, or for the CS to acquire predictive properties that signal reward availability.