Ethanol induces conditioned place preference in genetically selected alcohol-preferring rats

The study of the biological mechanisms of ethanol reward has greatly suffered from problems to obtain ethanol-induced conditioned place preference (CPP) in rats. In the present study, CPP was obtained in genetically selected Marchigian Sardinian alcohol-preferring (msP) rats, derived from Sardinian alcohol-preferring rats, following intragastric (IG) ethanol administration by means of a permanent IG catheter, but not after intraperitoneal (IP) injection or IG gavage. Rats with permanent IG catheter, received IG administration of 0.35, 0.7, 1.5 or 2.8 g/kg ethanol, as a 10% v/v solution. In ethanol-experienced rats 0.7 or 1.5, but not 0.35 or 2.8 g/kg ethanol significantly increased in comparison to controls the time spent in the ethanol-associated previously non-preferred compartment, which became preferred in the post-conditioning test. In ethanol-naive rats, only 0.7 g/kg ethanol significantly increased the time spent in the ethanol-associated compartment. On the other hand, no effect was observed in alcohol-experienced rats following IG gavage, or IP injection of 0.35, 0.7 or 1.5 g/kg ethanol. The present results provide evidence that ethanol possesses postingestive rewarding properties in msP rats, and that it can reliably induce CPP in them, provided that an appropriate method of administration is adopted. Received: 26 October 1997/Final version: 26 May 1998     

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.     

Antidepressant-like effect of ethanol revealed in the forced swimming test in Sardinian alcohol-preferring rats

  Rationale: A large body of evidence indicates high comorbidity between depression and alcohol abuse. The self-medication hypothesis proposes that depressed subjects may abuse ethanol because it reduces the symptoms of depression. The present study evaluated whether ethanol may exert an antidepressant-like action in genetically selected alcohol-preferring rats, either Sardinian alcohol-preferring (sP) or Marchigian Sardinian alcohol-preferring (msP) rats, and for comparison in Sardinian alcohol-non-preferring (sNP) rats. Methods: The forced swimming test (FST) was used to evaluate the antidepressant-like action of ethanol; in this test the effect of ethanol ingestion on the immobility time was determined. Results: Ethanol-naive sP rats exhibited a longer period of immobility in comparison to sNP rats. Both in ethanol-naive sP and msP rats, voluntary ethanol drinking reduced the immobility time. A similar effect was obtained when repeated (five or nine) intragastric administrations of 0.7 g/kg ethanol were given during the 24 h prior to the test in msP and in sP, but not in sNP rats. Desipramine, like ethanol, sharply reduced immobility at doses of 5 or 20 mg/kg, given 3 times in the 24 h before the test in msP rats. The reduced immobility induced by ethanol in msP rats was apparently not the consequence of a general motor activation, because 9 IG administrations of ethanol, 0.7 g/kg, failed to alter locomotor activity in the open field test. Moreover, blood alcohol levels and rectal temperature of msP, sP and sNP after IG ethanol administration were not statistically different. Conclusions: The present results provide evidence for an antidepressant-like action of ethanol in sP and msP rats and suggest that this action may contribute to sustain their high ethanol drinking. Received: 3 August 1998 / Final version: 22 December 1998     

Differences in response to the aversive properties of ethanol in rats selectively bred for oral ethanol preference

A conditioned taste aversion (CTA) paradigm was used to determine whether aversion to the pharmacological effects of ethanol, apart from orosensory cues, can contribute to genetic differences in voluntary ethanol consumption. Four doses of ethanol, administered IP, were paired with the consumption of a 0.1% saccharin solution in rats from the alcohol-preferring (P) and alcohol-nonpreferring (NP) lines. Repeated pairing of saccharin and ethanol in a dose of 1.0 g/kg produced stronger and more prolonged aversion to saccharin in NP rats, compared with P rats, at comparable blood ethanol levels. A low dose of ethanol (0.25 g/kg) produced transient conditioned facilitation of saccharin consumption in P rats, but not in NP rats, at comparable blood ethanol levels. The results suggest that rats of the NP line find the postingestional effects of high-dose ethanol more aversive, and low-dose ethanol less reinforcing, than do rats of the P line. Genetic differences in voluntary ethanol consumption may be due, in part, to differences in aversion to the postingestional effects of ethanol.     

Chronic alcohol consumption in alcohol-preferring P rats attenuates subsequent conditioned taste aversion produced by ethanol injections

Rats of the P line were tested for the development of tolerance to the aversive effects of ethanol during 33 days of continuous availability of food, water and a 10% (v/v) ethanol solution. Beginning on the day following the removal of ethanol, five daily conditioned taste aversion (CTA) trials were administered to the ethanol-drinking P rats and an ethanol-naive control group. The CTA trials consisted of a 20-min access to a Polycose solution, followed by IP injection of saline, 0.5, 1.0, or 1.5 g ethanol/kg. The ethanol-drinking rats developed a preference for the Polycose solution when it was paired with 0.5 g ethanol injections, but the control rats did not. Both control and ethanol groups had similar CTAs at the 1.5 g dose. However, at the 1.0 g dose, the ethanol group had an attenuated CTA compared with the water control group. The results suggest that P rats develop tolerance to aversive effects of ethanol during chronic drinking. This tolerance could contribute to the high ethanol intake in these selectively-bred rats.     

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.     

Reduction of ethanol intake by chronic treatment with Hypericum perforatum, alone or combined with naltrexone in rats

Acute treatment with extracts of Hypericum perforatum, the common plant usually called St. John's Wort, reduces voluntary ethanol intake in Marchigian Sardinian alcohol-preferring (msP) rats and acts synergistically with opioid receptor antagonists to further attenuate ethanol consumption. The present study evaluated the effect of chronic (once a day for 12 days) intragastric administration of a CO2 Hypericum perforatum extract (HPCO2), given alone or combined with naltrexone (NTX), on ethanol intake offered 2h/day in msP rats. Chronic treatment with HPCO2 markedly reduced ethanol intake at the dose of 125, but not at 7 mg/kg; the effect of 125 mg/kg was observed since the first day of treatment and remained constant across the 12 days. The same dose of HPCO2 slightly reduced the simultaneous intake of food only on day 3 and day 11 of treatment. Treated rats promptly recovered baseline ethanol intake when treatment did not precede access to ethanol (on day 8) or after the end of treatment (day 13 and day 14), suggesting that HPCO2 administrations did not induce conditioned aversion to alcohol. Chronic intraperitoneal treatment with NTX reduced ethanol intake at 3, but not at 0.5mg/kg. The synergistic effect on ethanol intake of HPCO2 and NTX was evident also in conditions of chronic treatment. HPCO2, 7 mg/kg, and NTX, 0.5mg/kg, evoked a pronounced and statistically significant reduction of ethanol intake, while being inactive. The effect on ethanol intake of the combined treatment remained stable over the 12 days of treatment; food intake was slightly reduced only on day 3 and on day 7 in response to 125 mg/kg of HPCO2 combined with NTX 0.5mg/kg, but no difference in body weight between controls and treated rats was observed at the end of treatment. Following 12-day treatment with 125 mg/kg of HPCO2, no difference was observed in the responsivity of msP rats to the effect on ethanol intake of several doses of the extract. In conclusion, the present results provide evidence for a selective and pronounced effect of HPCO2, alone or combined with naltrexone, on ethanol intake in conditions of chronic treatment, without development of tolerance. These findings further support the view that clinical trials for extracts of Hypericum perforatum in the treatment of alcoholism should be considered.     

Non-specific enhancement of ethanol-induced taste aversion by naloxone

The conditioned taste aversion paradigm (CTA) was used to examine the effects of naloxone on ethanol-induced aversion towards a saccharine solution (3 conditioning and 11 extinction trials). Six groups of rats received conditioning trials consisting of two IP injections after saccharine presentation of different combinations of either ethanol (E: 1.75 g/kg), LiCl (L: 12 mEq/kg, 0.1 M), naloxone (N: 10 mg/kg) or saline (S); S-S, S-N, E-S, E-N, L-S and L-N. Naloxone by itself produced no aversion to the saccharin flavor. Based on the onset and extinction of aversion, naloxone significantly enhanced ethanol but also LiCl-induced CTA. The comparative data argues in favor of different mechanisms of action (1) between the aversive central effects of ethanol and morphine and (2) between ethanol's acute behavioral effects and negatively reinforcing properties. Enhancement of ethanol and LiCl-induced CTA by naloxone is compatible with hypernociceptive action of the opiate-antagonist and with the pain-modulating role of opiates in the CNS.     

Cross-tolerance between delta9-tetrahydrocannabinol and ethanol: the role of drug disposition.

Acute challenge doses of delta9-tetrahydrocannabinol (THC), 10.1 mg/kg, administered intragastrically by gavage (IG), or ethanol, 1.24 g/kg, IP, reduced the rotarod performance of female rats by 50%. Daily treatment of the animals with THC, 10.1 mg/kg, IG, or ethanol, 4 g/kg, IG, resulted in tolerance development to the impairing effects of the challenge doses of each drug on rotarod performance. THC-tolerant animals were cross-tolerant to the challenge dose of ethanol, but ethanol-tolerant rats did not show complete cross-tolerance to the challenge dose of THC. THC-tolerant animals initially had higher blood levels of 14C-THC than controls after IG drug administration. Following IV injection, the rates of 14C-THC disappearance were equivalent in the latter groups. 14C-THC disappearance was not altered in ethanol-tolerant animals. The rates of ethanol disappearance were not significantly modified in THC- or ethanol-tolerant animals. In conclusion, THC-tolerant female rats demonstrated cross-tolerance to ethanol as shown previously for males. Furthermore, the development of tolerance and cross-tolerance was not a function of changes in drug disappearance.     

Behavioral and enzymatic interactions between benzyl alcohol and ethanol.

Acute IP injection of benzyl alcohol but not benzaldehyde (0.5 g/kg) caused aversion to voluntary drinking of 5% ethanol solution by male rats with preference to ethanol. Benzyl alcohol noncompetitively inhibited hepatic alcohol dehydrogenase of rats maintained for a short term on 5% ethanol compared to control. The results suggest an adverse interaction between benzyl alcohol and ethanol underlying the observed aversion to ethanol.     

Nicotine-induced conditioned taste aversion in the rat: effects of ethanol

It has been shown that small doses of ethanol antagonise the discriminative stimulus properties of nicotine in the rat. The aim of the present study was to evaluate whether ethanol could antagonise the aversive stimulus effects of nicotine. Wistar rats were trained to associate nicotine injections with a novel tasting fluid (0.1% saccharin) in the conditioned taste aversion procedure. Nicotine (0.3 mg/kg, s.c.) was injected 5 min after the end of a 20-min exposure to the saccharin solution. Ethanol (0.25-0.5 g/kg, i.p.) was administered 5 or 50 min before nicotine. In general, ethanol did not inhibit nicotine-induced conditioned taste aversion. Contrary to the findings in drug discrimination studies, a slight but significant enhancement of nicotine-induced taste aversion conditioning was observed after ethanol pre-treatment. Blood ethanol levels were measured in a separate group of rats. Maximal blood ethanol levels after i.p. administration of 0.25 or 0.5 g/kg ethanol exceeded 20 and 80 mg%, respectively. Concluding, the present results may indicate that ethanol does not attenuate nicotine-induced conditioned taste aversion in the rat.     

Effect of amygdaloid lesions on ethanol intake in rats

The effect of electrolytic lesions of the amygdala on ethanol intake in ethanol naïve rats has been studied. Rats with basolateral nuclei and lateral nuclei lesions showed a reduced neophobic response to an ethanol solution. However, the ethanol intake was too small in normal and lesioned rats to augment aversion through conditioning. Oral intake of ethanol supplemented by intraperitoneal ethanol injection to reach 2 g/kg indeed enhanced the initial sensory aversion to ethanol. This induced aversion was attenuated after basolateral lesions. An initial aversion to a mixed ethanol-sucrose solution was abolished after basolateral lesions, while the lateral lesions induced an initial preference for this solution. The initial oral intake of ethanol-sucrose in normal rats was again too small to induce the conditioned taste aversion (C.T.A.). Despite the high oral intake of this solution, rats with basolateral lesions did not show a conditioned aversion while laterally lesioned rats exhibited a strong conditioned aversion to the ethanol-sucrose mixture. The results which confirm the suppression of the C.T.A. by basolateral amygdala lesions are discussed in relation to the role of toxicophobia in ethanol intake by rats.     

Reinforcement with intragastric infusions of ethanol: blocking effect of FLA 57

Suppression of oral intake of ethanol by FLA 57 has been reported for rats and was attributed to an inhibition of dopamine beta-hydroxylase. We have demonstrated the ability of FLA 57 (50 mg/kg, IP) to suppress bar-pressing for intragastric (IG) delivery of doses of ethanol (25 mg/kg). This indicates that the effect on oral intake of ethanol may not be attributed to a taste factor, e.g., a decreased palatability of the ethanol solution. The same dose of FLA 57 did not suppress responding for IG doses of sweet milk. Thus, there was not an impairment of appetitive behavior in general through some nonspecific depressant or toxic action. Furthermore, the primary reinforcing action of ethanol, when used to establish a buzzer as a conditioned reinforcer through repeated pairings, was blocked if FLA 57 was given before pairings. This was evidenced by a failure of such rats to bar-press above the baseline level in a later test of conditioned reinforcement, which contrasted with the increased responding seen for rats receiving saline instead of FLA 57 before ethanol. These data support the previous findings on oral ethanol and confirm that FLA 57 can impair the mechanism by which ethanol produces positive reinforcement in rats.     

Attenuation and cross-attenuation in taste aversion learning in the rat: studies with ionizing radiation, lithium chloride and ethanol

The preexposure paradigm was utilized to evaluate the similarity of ionizing radiation, lithium chloride and ethanol as unconditioned stimuli for the acquisition of a conditioned taste aversion. Three unpaired preexposures to lithium chloride (3.0 mEq/kg, IP) blocked the acquisition of a taste aversion when a novel sucrose solution was paired with either the injection of the same dose of lithium chloride or exposure to ionizing radiation (100 rad). Similar pretreatment with radiation blocked the acquisition of a radiation-induced aversion, but had no effect on taste aversions produced by lithium chloride (3.0 or 1.5 mEq/kg). Preexposure to ethanol (4 g/kg, PO) disrupted the acquisition of an ethanol-induced taste aversion, but not radiation- or lithium chloride-induced aversions. In contrast, preexposure to either radiation or lithium chloride attenuated an ethanol-induced taste aversion in intact rats, but not in rats with lesions of the area postrema. The results are discussed in terms of relationships between these three unconditioned stimuli and in terms of implications of these results for understanding the nature of the proximal unconditioned stimulus in taste aversion learning.     

Effects of route of administration of ethanol on high-speed reaction time in young and old rats

The blood ethanol concentrations (BECs) and reactive capacity of young (8 months) and old (24 months) male Fischer 344 rats were compared at 5, 10, 20, 45, 65, and 90 min following the administration of ethanol (EtOH). The time-dependent effects of intragastric intubation (IG; 3 g/kg) and intraperitoneal injection (IP; 1.5 g/kg) of EtOH (20% w/v) were determined. Subsequent to IG delivery, BECs rose most rapidly within the first 20 min, but did not reach peak levels until 90 min for both young (240 mg/dl) and old rats (250 mg/dl). Following IP injections, BECs escalated within 5 min to 250 mg/dl in the young, to 175 mg/dl in the old, and declined gradually to a stabilized value of 150 mg/dl (young) and 130 mg/dl (old). The old rats never achieved the high BECs seen in the young. Reactive capacity, a measure of auditory/visual reaction time, was inversely related to BECs. As BECs (IP) declined, performance improved at a similar rate in both age groups, although the old rats' performance was more impaired than that of the young. However, BEC per se was not an adequate predictor of reactive capacity. When EtOH was delivered by IG so that BECs remained high for long periods of time, reactive capacity was far less impaired compared with IP delivery in which comparable BECs were present for only a few minutes. The possibility was noted that behavioral tolerance may have developed during the 90-min post-EtOH period, and that the IP delivery method may disrupt behavior in ways independent of brain ethanol levels.     

Motivational properties of ethanol in mice selectively bred for ethanol-induced locomotor differences

Ethanol-induced locomotor stimulation has been proposed to be positively correlated with the rewarding effects of ethanol (Wise and Bozarth 1987). The present experiments provided a test of this hypothesis using a genetic model. Three behavioral indices of the motivational effects of ethanol (drinking, taste conditioning, place conditioning) were examined in mice from two independent FAST lines, selectively bred for sensitivity to ethanol-induced locomotor stimulation, and mice from two independent SLOW lines, selectively bred for insensitivity to ethanol-induced locomotor stimulation. In a single-bottle procedure, mice were allowed access to drinking tubes containing ethanol in a concentration (1–12% v/v) that increased over 24 consecutive days. FAST mice consumed greater amounts of ethanol solution. In a two-bottle procedure, mice were allowed access to tubes containing water or various concentrations of ethanol (2–8% v/v) over 6 days. FAST mice generally showed greater preference for ethanol solutions than SLOW mice. In a conditioned taste aversion procedure, mice received access to saccharin solution followed by injection of 2.5 g/kg ethanol (IP). SLOW mice developed aversion to the saccharin flavor more readily than FAST mice. In a series of place conditioning experiments, tactile stimuli were paired with various doses of ethanol (0.8–2.0 g/kg). During conditioning, FAST mice showed locomotor stimulation after 1.0, 1.2 and 2.0 g/kg ethanol while SLOW mice did not. During testing, mice conditioned with 1.2 g/kg and 2.0 g/kg ethanol showed conditioned place preference, but there were no line differences in magnitude of preference. These results indicate that genetic selection for sensitivity to ethanol-stimulated activity has resulted in genetic differences in ethanol drinking and ethanol-induced conditioned taste aversion but not ethanol-induced conditioned place preference. Overall, these data provide mixed support for the psychomotor stimulant theory of addiction.     

Monoamine and metabolite levels in CNS regions of the P line of alcohol-preferring rats after acute and chronic ethanol treatment

Levels of norepinephrine (NE), dopamine (DA), 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA), serotonin (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) were determined in 8 brain regions of the P line of alcohol-preferring rats following: (a) an IP injection of 2.5 g ethanol/kg body wt; (b) 8 and 15 weeks of chronic free-choice drinking of 10% ethanol; (c) 15 weeks of chronic free-choice drinking of 10% ethanol and 24 hours of withdrawal; and (d) 7 weeks of forced administration of 5% ethanol in liquid diet. One hour after IP injection of 2.5 g ethanol/kg body wt, the levels of DOPAC and HVA increased 209-45% in the cerebral cortex (CTX) and striatum (STR). A 209% lower content of NE in the CTX of the ethanol group was the only other statistically significant difference observed. Chronic free-choice drinking of 10% ethanol for 8 weeks (6.5 +/- 0.4 g ethanol/kg/day) or 15 weeks (7.8 +/- 0.2 g ethanol/kg/day) and the chronic forced administration of ethanol in liquid diets (up to 13.2 +/- 0.2 g ethanol/kg/day) did not produce any consistent pattern of alterations in the levels of the monoamines or their metabolites in the 8 CNS regions. After 15 weeks of chronic free-choice drinking of 10% ethanol, withdrawal from alcohol also did not produce alterations in the content of the monoamines or their metabolites. These data indicate that acute administration of hypnotic doses of ethanol increases the metabolism of specific dopaminergic neurons in the CNS of the P rat, but monoamine levels and metabolism are not altered after chronic (7-15 weeks) alcohol consumption.     

Injection timing determines whether intragastric ethanol produces conditioned place preference or aversion in mice

Previous studies have shown that mice develop conditioned place preference (CPP) when ethanol is administered by intraperitoneal (ip) or intravenous (iv) injection. The present studies examined CPP in mice using the intragastric (ig) route of administration. Inbred mice were surgically implanted with chronic intragastric cannulae and exposed to an unbiased place conditioning procedure in which infusion of ethanol (2 or 4 g/kg) was paired with a conditioned stimulus (CS+). A different CS was paired with water. In Experiments 1-2, ethanol was infused just before exposure to CS+. Contrary to previous studies involving intraperitoneal injection, infusion of 4 g/kg ig ethanol produced a significant conditioned place aversion (CPA). However, when a 5-min delay was inserted between infusion and CS exposure (Experiments 3-4), the same dose produced CPP. These outcomes are not consistent with expectations derived from a recent study in selectively bred rats, suggesting that sensitivity to ethanol reward is enhanced by intragastric administration. However, the finding that intragastric ethanol can produce either CPP or CPA depending on dose and injection timing is consistent with previous intraperitoneal ethanol studies in mice. Although the parameters differ for each route of administration, it appears that the same underlying processes can be invoked to explain how manipulation of injection timing affects the direction of ethanol-induced place conditioning. More specifically, in both cases, CPA can be attributed to an initial, short-lived aversive effect, whereas CPP can be attributed to a delayed rewarding effect of ethanol.     

A homotaurine derivative reduces the voluntary intake of ethanol by rats: are cerebral GABA receptors involved?

The effects of some derivatives of homotaurine (3 APS), the well known GABA agonist, were tested on the voluntary intake of ethanol by rats. Spontaneously ethanol drinking rats (DR) were selected and had a constant voluntary intake of ethanol by rats. Spontaneously ethanol drinking rats (DR) were selected and had a constant voluntary intake of a 12% ethanol solution (VIE) during 14 days (about 5 g/kg body weight daily). Calcium acetylhomotaurine (0.26 and 0.52 mmol/kg daily IP) significantly reduced VIE and this was inhibited by the GABA antagonist bicuculline (2 mg/kg IP). The conditioned aversion test to saccharin was negative. Bicuculline alone did not affect VIE. Other homotaurine (3-APS) derivatives: sodium acetyl homotaurine (Na AOTA), homotaurine (OTA), sodium acetyltaurine (Na A TA) and calcium chloride (CaCl2) did not affect VIE. These data suggest that the gabaergic system could be implicated in VIE. MERAM Lab. patent.     

Ontogenetic difference in ethanol reinforcing properties: the role of the opioid system

Previous data indicate that ethanol intoxication (3 g/kg, intragastric) on postnatal day (PD) 7 and 8 increases ethanol acceptance, but on PD 10 and 11 generates an aversion in infant rats. We investigated the participation of the opioid system in these effects. Subcutaneous administration of naloxone (1 or 10 mg/kg) followed by ethanol intoxication on PD 7 and 8 prevented the increased ethanol intake effect observed in the younger pups, but when ethanol intoxication occurred on PD 10 and 11, naloxone treatment did not affect the aversion observed at this age. An aversion to ethanol was evidenced in the younger pups administered ethanol and naloxone, but only when exposed to ethanol odor during ethanol intoxication. Results indicate that the increased ethanol acceptance induced by ethanol intoxication in the younger pups is mediated by the opioid system, and that ethanol may also induce conditioned aversions at this early age.