Oral Ethanol-Reinforced Responding in Rhesus Monkeys: Effects of Opioid Antagonists Selective for the μ-,κ-, or δ-Receptor

To determine the mechanism by which naltrexone (NTX) reduces oral ethanol-reinforced responding, opioid antagonists that show μ-, κ-, or δ-selectivity were evaluated. Rhesus monkeys ( n = 6) were given opportunities to respond and receive ethanol (1 % or 2%) or water during daily 3-hr drinking sessions. Before some drinking sessions, the monkeys received intramuscular injections of saline or the following drugs: the μ-selective irreversible antagonist clocinnamox (CCAM), the κ-selective long-lasting antagonist nor-binaltorphimine (nor-BNI), or the δ-selective antagonist naltrindole. Also, NTX was administered along with either CCAM or nor-BNI. When given alone, CCAM (0.1 mg/kg) had no effect on ethanol-reinforced responding. When NTX (0.32 mg/kg) was given with CCAM, responding maintained by ethanol was decreased. Nor-BNI (3 mg/kg) reduced ethanol-reinforced responding only on the day of injection. On subsequent days, when other studies report continued κ-antagonism, responding maintained by ethanol returned to control levels. Also, NTX (0.32 mg/kg), administered in the presence of nor-BNI, was still able to reduce ethanol-reinforced responding. Naltrindole failed to alter responding maintained by ethanol. Because selective antagonism at the μ-, κ-, or δ-receptor did not reduce ethanol-reinforced responding, NTX's ability to reduce ethanol consumption may not be mediated by these previously characterized opioid receptors. NTX may exert its effects through an uncharacterized opioid binding site or through a nonopioid mechanism.     

Periodic Naltrexone and Propensity To Take Alcoholic Beverage

For over 2 months, 45 rats were maintained on a daily regimen involving 2 hr a day of access to both water and a palatable alcoholic beverage. At first, they took little ethanol. As days progressed, they eventually took over 2 g/kg of ethanol during the 2 hr. Previous research indicates that, without intervention, they would maintain this level of intake indefinitely. All rats were taken off the daily regimen for 30 days and then returned to it, i.e., rats received 30 days of "abstinence." For 35 days following abstinence, one-third of the subjects received placebos daily, one-third received naltrexone (NTX), 10 mg/ kg, daily, and the one-third received NTX on days l-5, ll-15,21-25, and 3135 and placebos on the other days. Abstinence reduced all rats' intakes of alcohol compared with pre-abstinence levels. Rats that received only placebos quickly returned to taking alcohol at pre-abstinence levels. Rats that received NTX daily increased their intakes up to the level normally expected for receiving NTX and no abstinence. Because rats receiving daily NTX always drank a fraction of the alcohol consumed by those receiving placebos, NTX's effects did not diminish. As rats sampled alcoholic beverage, however, the effects of abstinence did diminish. The rats of periodic NTX drank as rats getting NTX when they were given NTX and as rats getting placebos when they were given placebos. Furthermore, the rats of periodic NTX showed no carry-over effects from periods of NTX to no NTX. Abstinence and NTX together, apparently, reduce propensity to take alcoholic beverage more than either alone.     

Effects of MDL 72222, a serotonin3 antagonist, on operant responding for ethanol by alcohol-preferring P rats

BACKGROUND: Previous studies indicated that, under continuous access conditions, the 5-HT3 antagonist MDL 72222 (MDL) effectively reduced ethanol drinking of alcohol-preferring P rats. However, MDL was without effect when similar doses were tested under scheduled access conditions, unless the ethanol access period was randomly presented. This study examined the effects of MDL on operant responding for ethanol and water by adult male alcohol-preferring P rats. METHODS: During the dark cycle, subjects in the first experiment were trained to respond concurrently for 15% ethanol and water on a fixed-ratio 5 (FR-5) and FR-1 schedule of reinforcement, respectively. Approximately 30 min before the 4-hr operant session, rats were injected subcutaneously (sc) with saline or MDL (1, 3, or 5 mg/kg). A second experiment tested the effects of 1 mg/kg MDL on operant responding for 15% ethanol in 1-hr sessions when operant access was given at a fixed time each day (fixed scheduled access, FSA group) or at variable time periods throughout the dark cycle (variable scheduled access, VSA group). RESULTS: In the first experiment, only the 5 mg/kg dose of MDL decreased responding for ethanol (approximately 20%) during the first 30 min of the 4-hr session. This dose also reduced total 4-hr responding for ethanol and water. In the second experiment, the 1 mg/kg dose of MDL had no effect on operant responding by the FSA group, but significantly reduced ethanol responding by the VSA group. CONCLUSIONS: These results suggest that 5-HT3 receptors may be involved in mediating the reinforcing effects of ethanol, and that temporal-environmental cues associated with the presentation of ethanol may be one factor involved in reducing the effectiveness of 5-HT3 antagonists to attenuate ethanol intake.     

The Reinforcing Effects of Ethanol Are Altered by the Endogenous Neurosteroid, Allopregnanolone

We examined the effect of systemic administration of the endog-enously occurring progesterone metabolite, allopregnanolone, on oral self-administration of ethanol by male rats. Rats were trained to perform an operant response for presentation of 0.1 ml of a solution of 10% ethanol in water using the sucrose fading technique. After acquisition of stable lever-press responding on a fixed-ratio 4 schedule, subjects received subcutaneous injections of 1,3, or 10 mg/kg of allopregnanolone, or vehicle, 20 min prior to the self-administration session. Pretreatment with 3 mg/kg, but not 1 or 10 mg/kg, increased the mean total number of lever press responses made to obtain ethanol, and therefore increased the mean total number of ethanol presentations. The number of responses and response rate were examined as a function of the number of “runs” within the 30-min session; a “run” was defined as a series of consecutive responses with an interresponse interval of <1 min. The increase in total responses after 3 mg/kg was due in part to an increased number of responses for the first run of the session, with no effect on response rates. However, the higher dose of 10 mg/kg decreased response rates within the first run. Thus, allopregnanolone alters ethanol-reinforced responding at concentrations lower than those that depress rates of responding. The effects of administration of the ben-zodiazepene, diazepam, were determined for comparison with those of the neurosteroid. The subcutaneous injection of 0.3, 1.0, or 3.0 mg/kg of diazepam did not produce any clear dose-dependent changes in measures of ethanol-reinforced operant responding, supporting the suggestion of differences in the contribution of the benzodiazepene and neurosteroid binding sites to GABA_A receptor function. The results indicate that exogenous administration of allopregnanolone dose-dependently alters ethanol-reinforced operant responding, and suggest that this endogenously occurring neurosteroid could mediate some of the reinforcing effects of ethanol.     

Responding for Oral Ethanol after Naloxone Treatment by Alcohol-Preferring AA Rats

The present study examined the effect of a relatively nonselective opioid antagonist, naloxone, on lever pressing for oral ethanol by the alcohol-preferring AA rats. The AAs, housed continually in operant chambers with free access to food and water, learned to respond for 10% oral ethanol during daily 60-min alcohol access periods indicated by a stimulus light. The rats developed stable ethanol responding, resulting in mean ethanol intakes of 1.2 g/kg/60 min and measurable blood alcohol levels. In the first experiment, single systemic injections of naloxone (0.05–2.5 mg/kg) had no effect on the initial rate of responding; dose-dependent decreases were observed later during the alcohol access. The second experiment examined the effects of repeated injections of 0.5 and 2.5 mg/kg naloxone on 5 consecutive days. Naloxone suppressed responding dose-relatedly over the treatment days. In contrast to the effects of single injections, repeated injections with 2.5 mg/kg naloxone produced progressive decreases within the first minutes of access. The results suggest that naloxone may attenuate the reinforcing actions of ethanol.     

Nitric oxide synthesis inhibition attenuates conditioned reinstatement of ethanol-seeking, but not the primary reinforcing effects of ethanol

Background: Nitric oxide (NO) signaling has been implicated in regulating aspects of the reinforcing and addictive actions of cocaine. These experiments were designed to examine whether NO-dependent neurotransmission also participates in mediating the addictive actions of another drug of abuse, ethanol, with emphasis on both the primary reinforcing effects of ethanol and the incentive motivational effects of ethanol-related contextual stimuli.
Methods: Male Wistar rats were operantly trained to orally self-administer 10% (w/v) ethanol in daily 30-min sessions and to associate distinct discriminative stimuli with the availability of ethanol (S⁺) versus nonreward (S⁻). Rats were treated with the NO synthase inhibitor N ^G-nitro-l-arginine methyl ester (l-NAME; 0, 10, or 40 mg/kg intraperitoneally) 30 min before self-administration tests that were conducted after establishment of stable levels of daily ethanol intake and conditioned reinstatement tests that were performed after extinction of ethanol-maintained operant responding.
Results: l-NAME did not alter the primary reinforcing effects of ethanol in self-administration tests. In contrast, l-NAME dose-dependently attenuated the recovery of extinguished responding induced by the ethanol S⁺ in the absence of ethanol availability during reinstatement tests.
Conclusions: These results suggest that the NO system does not play a role in behavior reinforced directly by ethanol. However, the results implicate NO-dependent neurotransmission in alcohol-seeking responses elicited by drug-related contextual stimuli.     

Naltrexone Blocks Acquisition of Voluntary Ethanol Intake in Rats

The effects of naltrexone (NTX) on the acquisition of ethanol drinking was assessed in rats. NTX (0, 2.5, 5.0, or 10.0 mg/kg) was administered to rats presented with an ascending series of ethanol concentrations (2%, 4%, 6%, and 8% v/v) and water. The 2.5 and 10 mg/kg doses of NTX attenuated the acquisition of voluntary drinking of 8% ethanol, but the 5.0 mg/kg dose of NTX had no effect on ethanol intake. The acquisition paradigm was repeated in experiment 2 with naïve animals that received 0, 5.0, or 7.5 mg/kg of NTX. Neither dose of NTX affected ethanol intake, preference for alcohol, or water intake. Total fluid intake was suppressed in the NTX groups, but only on the second presentations of the 2% and 6% concentrations of ethanol. We suggest that the 2.5 and 10 mg/kg doses of NTX may have attenuated the acquisition of ethanol drinking by at least two different behavioral mechanisms.     

Suppression of ethanol responding by centrally administered CTOP and naltrindole in AA and Wistar rats

BACKGROUND: Both mu- and delta-opioid receptors have been implicated in the reinforcing actions of ethanol. However, selective opioid receptor antagonists have not altered ethanol intake in all rodent strains consistently, which suggests that genotype may modulate their suppressive effects. Therefore, we tested the effects of the selective mu-antagonist D-Pen-Cys-Tyr-D-Trp-Orn-Thr-Pen-Thr-NH2 (CTOP) and the selective delta-antagonist naltrindole in both high-drinking AA (Alko, Alcohol) and heterogeneous Wistar rats. METHODS: AA and Wistar rats were trained to respond for ethanol (10% w/v) in a two-lever operant condition by using a saccharin fading procedure. After stable baseline responding was established, rats were implanted stereotaxically either with a guide cannula above the lateral ventricle or with bilateral cannulas above the nucleus accumbens, basolateral amygdala, or ventral tegmental area. After postoperative recovery, AA and Wistar animals were tested after intracerebroventricular microinjections of either CTOP (0-3 microg) or naltrindole (0-30 microg) or subcutaneous injections of naloxone (0-1 g/kg), which was used as a reference antagonist. Effects of intracerebral microinjections of CTOP and naltrindole (both 0-500 ng) were tested only in Wistar rats. RESULTS: Subcutaneous naloxone and intracerebroventricular CTOP and naltrindole suppressed ethanol self-administration in a similar manner in AA and Wistar rats. Cumulative response patterns indicated that naloxone and naltrindole had no effect on the initiation of responding but suppressed it later during the session, whereas CTOP also affected initiation. In Wistar rats, naltrindole microinjections into both the nucleus accumbens and basolateral amygdala decreased ethanol responding, whereas CTOP was effective only in the amygdala. Injections of these antagonists into the ventral tegmental area had little effect on ethanol intake. CONCLUSIONS: The results confirm previous results which showed that both mu- and delta-opioid receptors are involved in the regulation of ethanol self-administration and indicate that genetic differences between AA and Wistar rats produced by selection do not modify the effects of opioid antagonists. The nucleus accumbens and the basolateral amygdala may be important central sites for the mediation of their suppressive effects.     

Effects of Morphine and Naloxone on Ethanol- and Sucrose-Reinforced Responding in Nondeprived Rats

In the following series of experiments, effects of morphine (0.1, 0.3, 1.0, 3.0, and 10.0 mg/kg) and naloxone (0.1, 0.3, and 1.0 mg/kg) were assessed in nondeprived rats trained to leverpress with 10% ethanol, sweetened ethanol, or 5% sucrose and water as the reinforcers. Morphine, at doses of 0.1, 0.3, and 1.0 mg/kg had little effect on responding with ethanol or sweetened ethanol available on a fixed ratio 4 (FR4) schedule of reinforcement, but at the 3.0 mg/kg dose, morphine suppressed responding to near zero. Similar results were obtained when 10% ethanol and water were available on a concurrent FR4 FR4 schedule of reinforcement. When 5% sucrose and water were available concurrently, morphine suppressed responding at 3.0 and 10 mg/kg. Naloxone (0.1, 0.3, and 1.0 mg/kg) decreased responding for ethanol, sweetened ethanol, and sucrose solutions in a dose-dependent manner. Naloxone decreased total number of responses/session by shortening the duration of responding without affecting momentary rate. Overall, the data suggest that the endogenous opioid system plays a role in the ability of ethanol to reinforce operant behavior. However, this role does not appear to be specific to ethanol because similar results were observed with sucrose reinforcement. Failure to find enhanced ethanol intakes following morphine injections in the operant situation suggests that the method used to measure ethanol self-administration makes a difference in assessing the effects of drugs on ethanol intake.     

Effects of L-Type Voltage-Sensitive Calcium Channel Modulators on the Discriminative Stimulus Effects of Ethanol in Rats

The purpose of the present investigation was to determine whether administration of dihydropyridine-sensitive calcium channels plays a role in modulating the discriminative stimulus effects of ethanol. A food-reinforced operant methodology was used to train adult male Long-Evans rats to discriminate either 1.0 g/kg of ethanol from water or 2.0 g/kg of ethanol from water. After training, two sets of experiments were conducted. First, a time course procedure was implemented whereby a single intraperitoneal dose of either nimodipine (3, 10, 30 mg/kg), nifedipine (3, 10, 30 mg/kg), or isradipine (1, 3, 10, 17 mg/kg) was administered, and test sessions were conducted 10, 20, 30, 60, and 90 min postinjection. Complete substitution (80% or greater ethanol-appropriate responding) for ethanol by these dihydropyridine compounds varied among subjects with dose and pretreatment time. Overall, isradipine substituted for the discriminative stimulus effects of ethanol in the greatest percentage of animals in both training groups. However, substitution varied with dose. Nifedipine dose dependently substituted for ethanol in half of the animals trained with 1.0 g/kg of ethanol but was less effective in animals trained with 2.0 g/kg of ethanol. For the second set of experiments, a single dose of nimodipine, nifedipine, isradipine, or (-)-BAY k 8644 was administered before determination of the cumulative ethanol dose response. Nifedipine produced a significant leftward shift and (-)-BAY k 8644 produced a significant rightward shift in the ethanol dose-response curve in animals trained to discriminate 2.0 g/kg of ethanol from water. These results indicate that the administration of VGCC modulators plays an indirect role in the discriminative stimulus effects of ethanol.     

Effects of Pregnanolone and Dehydroepiandrosterone on Ethanol Intake in Rats Administered Ethanol or Saline during Adolescence

Background: Adolescent alcohol use may contribute to long‐term changes in the receptors and neuroactive steroids that may mediate its effects and to subsequent alcohol abuse and dependence as an adult. Therefore, in this study, ethanol preference and intake as an adult were examined after adolescent ethanol or saline administration. In addition, ethanol intake in the same groups was examined after administration of 2 neuroactive steroids with modulatory effects at GABA_A receptors. Methods: Two groups of male Long‐Evans rats were administered 15 intraperitoneal (i.p.) injections of either ethanol (2 g/kg, 20% v/v) or saline between postnatal days 35 and 63. Starting on postnatal day 75, both groups were trained to consume 10% ethanol using a saccharin‐fading procedure, and ethanol intake and preference were measured after a series of manipulations involving food deprivation, changes in the duration of access to ethanol, and changes in the concentrations of ethanol presented. Following these manipulations, pregnanolone (1 to 10 mg/kg) and dehydroepiandrosterone (DHEA, 1 to 100 mg/kg) were administered prior to preference sessions with an 18% ethanol solution. Results: Adult ethanol preference and intake did not differ significantly in subjects treated with either saline or ethanol as adolescents during training, the substitution of other ethanol concentrations (3.2 to 32%), ad‐lib feeding, or moderate food deprivation. Pregnanolone administration altered the intake of both adolescent‐treated groups after the first injection of 3.2 mg/kg and after repeated injections with 10 mg/kg, a dose that produced sedation. In contrast, multiple doses of DHEA consistently decreased intake of an 18% ethanol concentration in both groups after repeated injections and 3 doses of DHEA (10, 32, and 56 mg/kg) administered with various ethanol concentrations dose‐dependently shifted the ethanol‐concentration curves for the volume and dosage of ethanol consumed downward. Conclusions: These results indicate that chronic intermittent ethanol (CIE) administration of 2 g/kg during adolescence did not alter preference or overall consumption of ethanol in outbred rats trained to drink ethanol as an adult under the conditions tested, and that DHEA may be more effective than pregnanolone at significantly decreasing ethanol consumption.     

Pregnenolone and ganaxolone reduce operant ethanol self-administration in alcohol-preferring p rats

Background: Neuroactive steroids modulate ethanol intake in several self‐administration models with variable effects. The purpose of this work was to examine the effects of the long‐acting synthetic GABAergic neurosteroid ganaxolone and the endogenous neurosteroid pregnenolone, a precursor of all GABAergic neuroactive steroids, on the maintenance of ethanol self‐administration in an animal model of elevated drinking—the alcohol‐preferring (P) rats. Methods: P rats were trained to self‐administer ethanol (15% v/v) versus water on a concurrent schedule of reinforcement, and the effects of ganaxolone (0 to 30 mg/kg, subcutaneous [SC]) and pregnenolone (0 to 75 mg/kg, intraperitoneal [IP]) were evaluated on the maintenance of ethanol self‐administration. After completion of self‐administration testing, doses of the neuroactive steroids that altered ethanol self‐administration were assessed on spontaneous locomotor activity. Finally, the effect of pregnenolone administration on cerebral cortical levels of the GABAergic neuroactive steroid (3α,5α)‐3‐hydroxypregnan‐20‐one (allopregnanolone, 3α,5α‐THP) was determined in both ethanol‐experienced and ethanol‐inexperienced P rats because pregnenolone is a precursor of these steroids. Results: Ganaxolone produced a dose‐dependent biphasic effect on ethanol reinforcement, as the lowest dose (1 mg/kg) increased and the highest dose (30 mg/kg) decreased ethanol‐reinforced responding. However, the highest ganaxolone dose also produced a nonspecific reduction in locomotor activity. Pregnenolone treatment significantly reduced ethanol self‐administration (50 and 75 mg/kg), without altering locomotor activity. Pregnenolone (50 mg/kg) produced a significant increase in cerebral cortical allopregnanolone levels. This increase was observed in the self‐administration trained animals, but not in ethanol‐naïve P rats. Conclusions: These results indicate that pregnenolone dose‐dependently reduces operant ethanol self‐administration in P rats without locomotor impairment, suggesting that it may have potential as a novel therapeutic for reducing chronic alcohol drinking in individuals that abuse alcohol.     

Combination of Naltrexone and Fluoxetine on Rats'Propensity to Take Alcoholic Beverage

Naltrexone (NTX) and fluoxetine (FLU) are useful for treating alcoholism and depression, respectively. Furthermore, these afflictions co-vary. Given the possibility that people might be prescribed NTX and FLU concurrently, we assessed the effects of these two agents on rats'propensity to drink an alcoholic beverage. Rats were given 66 days of access to a sweetened alcoholic beverage and water for 2 hr daily. At first, they took little ethanol, but after 20 days, they took on average 2.0 to 2.5 g/kg of ethanol, daily during the 2-hr session. They also took sufficient water to maintain their health. After 30 days, they were divided into four groups to receive, 30 min before the drinking session, 1 of 4 different kinds of injections. For the next 20 days, one group received placebo daily. Another group received 5 mg/kg of NTX dairy and another 5 mg/kg of FLU dairy. The fourth group received both 5 mg/kg of NTX and 5 mg/kg of FLU dairy. After 20 days, the doses of NTX and FLU were doubled across an additional 10 days. Both NTX and FLU reduced rats'intake of alcoholic beverage. The combinations of NTX and FLU, however, were no more effective in reducing rats'intake of alcoholic beverage than either alone. Also, the small dose of NTX seemed to lose its effectiveness with repeated administrations. A second experiment con-firmed the conclusion that small doses of NTX lose their effectiveness in suppressing intake of alcoholic beverage across repeated administrations. In summary, data provide no support for the idea that FLU and NTX would act synergistically to reduce propensity to take alcoholic beverages.     

Ethanol self-administration is regulated by CB1 receptors in the nucleus accumbens and ventral tegmental area in alcohol-preferring AA rats

Background: Endogenous cannabinoids and their receptors, CB1 receptors in particular, have been implicated in mediation of ethanol reinforcement. Previously, suppression of ethanol drinking by CB1 antagonists has been demonstrated in many experimental paradigms. However, the exact mechanism by which CB1 antagonists modulate ethanol drinking remains elusive. In the present study, we assessed the role of CB1 receptors within the key regions of the mesolimbic dopamine pathway, the nucleus accumbens (NAcc) and ventral tegmental area (VTA), in regulation of ethanol self‐administration. Methods: Adult male alcohol‐prefer AA rats were trained to self‐administer either 10% (w/v) ethanol or 0.1% (w/v) saccharin under an FR1 schedule during daily 30‐minute sessions. Following stable baseline responding, rats were tested after systemic administration of the CB1 antagonist SR141716A (0 to 10 mg/kg) and the agonist WIN55,212‐2 (0 to 2 mg/kg). Separate groups of rats were implanted with bilateral cannulas aimed at the NAcc or VTA, and tested after microinjections of SR141716A (0 to 3 μg) and WIN55,212‐2 (0 to 5 μg) into the NAcc or VTA. The highest intracerebral doses were tested also in rats responding for a 0.1% saccharin solution. Results: SR141617A dose‐dependently suppressed ethanol responding after systemic administration. Microinjections of SR141617A both into NAcc and VTA attenuated ethanol responding. In addition, intra‐NAcc injections of SR141617A suppressed saccharin intake. Although low doses of systemically given WIN55,212‐2 increased ethanol responding, no effects were seen after WIN55,212‐2 microinjections into NAcc or VTA. Conclusions: Bidirectional changes in ethanol self‐administration by the systematically administered CB1 agonist and antagonist show that ethanol reinforcement is controlled by CB1 receptors in alcohol‐preferring AA rats. Replication of the suppressive effects by CB1 antagonism in the NAcc and VTA suggests that endocannabinoids and their receptors mediate ethanol reinforcement through interaction with the mesolimbic dopamine pathway.     

Sex differences in non-reinforced responding for cocaine

Prior studies report no sex differences in cocaine consumption during maintenance of self-administration. We find female rats show poorer lever discrimination during acquisition of self-administration. Now, we test whether female rats show greater non-reinforced or ineffective responding (presses during infusion and time-out periods as well as inactive lever presses) than male rats during maintenance of cocaine self-administration (.0625-1.0 mg/kg/infusion) in Experiment 1. Persistence of responding during extinction when saline-replaced cocaine was also examined. Whether response differences reflect sex differences in movements under a non-drug condition was tested in Experiment 2. Because cocaine may affect lever press rates differentially between sexes, we examined the effects of cocaine (.3-30 mg/kg; IP) on responding for food in Experiment 3. Cocaine consumption does not differ between female and male rats. However, females respond more during infusion and time-out periods and during extinction than males. There is no sex difference in movements and high cocaine doses decrease responding for food more in female vs. male rats. That females engage in more ineffective responding may represent heightened "craving" and cannot be explained by increased movements or cocaine-stimulated increases in lever pressing. In contrast, responding for cocaine in males appears driven by drug delivery.     

Effects of benzodiazepines on acute and chronic ethanol-induced nociception in rats

BACKGROUND: Acute and chronic ethanol produces antinociception, and ethanol withdrawal induces hyperalgesia. METHODS: A radiant heat tail-flick assay was used to assess the effects of benzodiazepine ligands on ethanol-induced changes in nociception in rats. Acute activity of cumulative doses of ethanol (0.5-2.0 g/kg) and diazepam (0.1-10 mg/kg), a benzodiazepine agonist, was tested alone and after pretreatment with flumazenil (1.0-10 mg/kg), a benzodiazepine antagonist. Chronic effects of ethanol were tested in three groups of rats that received a liquid diet for 10 days. One group received ethanol alone; one group received ethanol and twice-daily injections of flumazenil (10 mg/kg); and one received a dextrin control diet. Acute withdrawal was tested at 12 hr after removal of the liquid diet. Effects of cumulative doses of diazepam (1.0-10 mg/kg) were tested during withdrawal (12 hr) in the ethanol-alone group. RESULTS: Acute doses of ethanol produced a small but significant degree of antinociception, which was fully suppressed by flumazenil. Acute doses of diazepam did not produce antinociception. Chronic exposure to ethanol produced antinociception on days 2 through 8. Tolerance developed by day 10, and hyperalgesia was seen 12 hr after removal of ethanol. Administration of diazepam or ethanol during withdrawal reversed the hyperalgesia induced by ethanol withdrawal. However, flumazenil (10-50 mg/kg) failed to reverse the antihyperalgesic effect of either diazepam or ethanol. No antinociception was seen in either the ethanol/flumazenil or dextrin control groups. CONCLUSIONS: These results suggest that the antinociceptive effects of both acute and chronic ethanol are at least partially mediated by GABA receptors, and that diazepam's antihyperalgesic effects may not be mediated by the GABA acid receptor.     

Role of acetaldehyde in the discriminative stimulus effects of ethanol

BACKGROUND: Acetaldehyde has been suggested to mediate some of the effects of ethanol. Acetaldehyde can be produced by the enzyme catalase within the brain after ethanol administration. The catalase inhibitor 3-amino-1,2,4-triazole (AT) reduces the production of acetaldehyde, and AT administration can reduce a number of ethanol-induced behavioral effects; this suggests the involvement of acetaldehyde in these behaviors. However, a role for acetaldehyde in mediating the discriminative stimulus effects of ethanol remains unclear. METHODS: The contribution of acetaldehyde to the discriminative stimulus effects of ethanol was investigated by use of a two-lever drug discrimination paradigm with food reinforcement. Male Long-Evans rats were trained to discriminate water from either 1.0 or 2.0 g/kg ethanol. Stimulus substitution tests were conducted with ethanol (0-2.5 g/kg by gavage) and acetaldehyde (0-300 mg/kg intraperitoneally). A cumulative dose-response procedure was then used to investigate the effects of pretreatments with AT (0.5 and 1.0 g/kg intraperitoneally) on ethanol discrimination. RESULTS: Acetaldehyde up to doses that decreased response rates (300 mg/kg) did not substitute for the discriminative stimulus effects of 1.0 or 2.0 g/kg ethanol. In addition, AT pretreatment did not affect the dose-response curves for ethanol discrimination. CONCLUSIONS: These results show that exogenous acetaldehyde administration does not produce discriminative stimulus effects that are similar to those of ethanol. Also, pretreatment with the catalase inhibitor did not affect the dose-response curve for ethanol discrimination, and this suggests that endogenously produced acetaldehyde does not contribute to the discriminative stimulus effects of ethanol. Together these results suggest that acetaldehyde does not mediate the discriminative stimulus effects of 1.0 to 2.0 g/kg ethanol.     

Interaction between family history of alcoholism and Locus of Control in the opioid regulation of impulsive responding under the influence of alcohol

Background: Naltrexone (NTX) is an opioid antagonist indicated for the treatment of alcoholism, which is not universally effective. Thus, identifying individual predictors of NTX’s behavioral effects is critical to optimizing its therapeutic use. Moreover, given the high rate of relapse during treatment for alcoholism, understanding NTX’s behavioral effects when combined with moderate ethanol intake is important. Our previous study of abstinent alcoholics and control subjects showed that a more internal Locus of Control score predicted increased impulsive choice on NTX ( Mitchell et al., 2007 , Neuropsychopharmacology 32:439–449). Here, we tested whether this predictive relationship remains in the context of moderate alcohol intake. Methods: In this study, we tested the effect of acute NTX (50 mg) on impulsive choice, motor inhibition, and attentional bias after ingestion of moderate ethanol (～0.3 g/kg, n = 30 subjects). Subjects included those recruited from a pool of ～1,200 UC Berkeley undergraduates on the basis of scores on the Barratt Impulsiveness Scale (BIS). Results: Impulsive choice was positively correlated with breath alcohol concentration in placebo sessions. Locus of Control was again the sole predictor of NTX’s effect on decision making among subjects with a family history of alcoholism. We also found a weak interaction between BIS scores and NTX’s effect on impulsive choice. Conclusions: Our results reinforce the predictive relationship between Locus of Control and NTX’s effect on decision making in those with a family history of alcoholism, suggesting a possible biological basis to this relationship.     

The gamma-aminobutyric acid-B receptor agonist baclofen attenuates responding for ethanol in ethanol-dependent rats

BACKGROUND: Gamma-aminobutyric acid-B (GABA(B)) receptor agonists have been shown to suppress operant self-administration of ethanol in nondependent rats. However, little work has focused on the effects of GABA(B) receptor agonists on self-administration of ethanol in dependent animals. METHODS: In the present experiment, the GABA(B) receptor agonist baclofen was tested for the ability to modulate both fixed- (FR) and progressive-ratio (PR) responding for ethanol in rats while nondependent and subsequently after ethanol dependence induction. Following the acquisition and stabilization of baseline operant ethanol self-administration and after dependence induction, baclofen [0.0, 0.5, 1, 2, and 4 mg/kg, intraperitoneal (IP)] was tested on FR-1 responding for ethanol. The ability of baclofen (2.0 mg/kg) to affect responding under a PR schedule of reinforcement was also evaluated. Dependence was induced in the animals by subjecting them to a 1-month intermittent vapor-exposure period in which animals were exposed to ethanol vapor for 14 h/d. Following the 1-month period, the vapor-exposed animals resumed FR-1 and PR baclofen drug testing (doses as described above) in the operant chambers at a time point corresponding to the animals being 6 hours into withdrawal (i.e., 6 hours after the ethanol vapor had been discontinued for that day). RESULTS: Baclofen (0.0, 0.5, 1, 2, and 4 mg/kg, IP) dose-dependently decreased ethanol self-administration in both nondependent and dependent rats on a FR schedule of reinforcement. However, the dose of baclofen that significantly reduced responding for ethanol was shifted to the left in the ethanol vapor-exposed animals, indicating an increased sensitivity to baclofen in animals that were chronically exposed to ethanol. When tested using a PR schedule of reinforcement, there was a significant increase in the breakpoint for the vapor-exposed animals (i.e., the animals were willing to work more in a dependent state). Baclofen (2.0 mg/kg, IP) suppressed intake for both nondependent and dependent animals. CONCLUSIONS: Ethanol dependence produced increased self-administration of ethanol as reflected in increased ethanol intake and increased responding on a PR schedule of reinforcement. As baclofen suppressed ethanol self-administration and showed evidence of increased potency in dependent animals, the present experiment suggests that the GABA(B) receptor could be a potential pharmacotherapeutic target for the treatment of chronic alcoholism.     

The Alcohol Deprivation Effect in C57BL/6J Mice is Observed Using Operant Self-Administration Procedures and is Modulated by CRF-1 Receptor Signaling

Background: The alcohol deprivation effect (ADE) is characterized by transient excessive alcohol consumption upon reinstatement of ethanol following a period of ethanol deprivation. While this phenomenon has been observed in rats using both bottle drinking (consummatory behavior) and operant self‐administration (consummatory and appetitive “ethanol‐seeking” behavior) procedures, ADE studies in mice have primarily relied on bottle drinking measures. Furthermore, the neurochemical pathways that modulate the ADE are not well understood. Therefore, we determined whether the ADE can be observed in C57BL/6J mice using operant self‐administration procedures and if expression of the ADE is modulated by the corticotropin releasing factor‐1 (CRF‐1) receptor. Methods: C57BL/6J mice were trained in a 2‐hour operant self‐administration paradigm to lever press for 10% ethanol or water on separate response keys. Between operant sessions, mice had access to ethanol in their homecage. Once stable responding occurred, mice were deprived of ethanol for 4 days and were then retested with ethanol in the operant paradigm for 3 consecutive days. Next, to assess the role of the CRF‐1 receptor, mice were given intraperitoneal (i.p.) injection (0, 10, or 20 mg/kg) of the CRF‐1 receptor antagonist CP‐154,526 30 minutes before ADE testing. Additional experiments assessed (i) ADE responding in which the alternate response lever was inactive, (ii) the effects of CP‐154,526 on self‐administration of a 1% sucrose solution following 4 days of deprivation, and (iii) ADE responding in which mice did not received i.p. injections throughout the experiment. Results: Mice exhibited a significant increase in postdeprivation lever responding for ethanol with either a water reinforced or inactive alternate lever. Interestingly, i.p. injection of a 10 mg/kg dose of CP‐154,526 protected against the ADE while not affecting lever responding for a sucrose solution. Finally, baseline and deprivation‐induced increases of ethanol reinforced lever responding were greater in mice not given i.p. injections. Conclusions: The ADE in C57BL/6J mice can be modeled using the operant self‐administration paradigm and increased ethanol self‐administration associated with the ADE is modulated by CRF‐1 receptor signaling.