Nicotinic receptor ligands reduce ethanol intake by high alcohol-drinking HAD-2 rats

Neuronal nicotinic acetylcholine receptors (nAChRs) are implicated in the reinforcing effects of many drugs of abuse, including ethanol. The present study examined the efficacy of cytisine, a nAChR partial agonist, and lobeline, a putative nAChR antagonist, on the maintenance of ethanol drinking by HAD-2 rats. Adult male HAD-2 rats were given access to ethanol (15 and 30%, with ad libitum access to water and food) 22 h/day for 12 weeks, beginning at 60 days of age, after which cytisine (0.0, 0.5, and 1.5 mg/kg) was tested for 3 consecutive days. The rats were given an 18-day washout period and were then tested with lobeline (0.0, 1.0, and 5.0 mg/kg) for 3 consecutive days. Ethanol intake was measured at 1, 4, and 22 h postinjection. Rats were injected intraperitoneally just before lights out (1200 h). There was a significant main effect of cytisine treatment on the second test day, with the 1.5 mg/kg dose significantly reducing ethanol intake at the 1- and 4-h time-points, relative to saline, and the 0.5 mg/kg dose inducing a significant reduction at the 4-h time-point. Conversely, lobeline treatment resulted in significant main effects of treatment for all three time-points within each test day, with the 5.0 mg/kg dose significantly reducing ethanol intake, relative to saline, at each time-point within each test day. These findings provide further evidence that activity at the nAChR influences ethanol intake and is a promising target for pharmacotherapy development for the treatment of alcohol dependence and relapse.     

Patterns of ethanol and saccharin intake in P rats under limited-access conditions.

Patterns of drinking and responding for ethanol (EtOH) and saccharin (SACC) were examined in the alcohol-preferring P rat using various limited-access paradigms. Adult female P rats (n = 10-20) were given 2-h access to EtOH (10-13% v/v) and SACC (0.0125% g/v) concurrently each day, or each solution individually on alternate days. Total 2-h SACC intake was significantly greater than EtOH under both concurrent (12+/-2 vs. 7+/-0.8 ml, p<0.05) and alternate-day access (18+/-1.6 vs. 10+/-0.5 ml) conditions. Under both conditions, however, EtOH intake (over 55% of the total) in the first 15 min was significantly greater than that of SACC (<25% of total). In an operant paradigm, total responding for EtOH (124+/-29) and SACC (114+/-7) under 2-h alternate-day conditions did not differ, but 65% of total EtOH responding occurred during the first 20 min versus less than 45% for SACC (p<0.05). Increasing response requirements (FR-1 to FR-5) did not significantly alter the total number of EtOH reinforcements, but decreased the total number of SACC reinforcements by approximately 50% (p<0.05). Increasing the EtOH concentration from 15% to 35% decreased the number of reinforcements approximately 50% but did not decrease the estimated g/kg EtOH intake. Increasing the SACC concentration from 0.0125% to 0.05%, however, nearly doubled the number of reinforcements. The greater preference for EtOH versus SACC during the initial part of the access period, together with the maintenance of EtOH intake in g/kg when the response requirements and the EtOH concentration were increased, suggests that EtOH intake is motivated by pharmacological consequences. Therefore, different motivational factors appear to underlie EtOH and SACC intake of the P rat. Furthermore, the pattern of EtOH intake and responding displayed by the P rat may be the result of a "bout-" or "binge-" like loss of control under restricted EtOH access conditions.     

Effects of 7-keto dehydroepiandrosterone on voluntary ethanol intake in male rats

Administration of dehydroepiandrosterone (DHEA), a neurosteroid that can negatively modulate the GABA_A receptor, has been shown to decrease voluntary intake of ethanol in rats. In vivo, DHEA can be metabolized to a variety of metabolites, including 3β-acetoxyandrost-5-ene-7,17-dione (7-keto DHEA), a metabolite without the prohormonal effects of DHEA. This study compared the effectiveness of 7-keto DHEA with DHEA for reducing ethanol intake in the same group of rats. The subjects, previously trained to drink ethanol using a saccharin-fading procedure, had access to ethanol for 30 min daily and the amount consumed was recorded. Subjects were administered 10 and 56 mg/kg of DHEA or 7-keto DHEA intraperitoneally 15 min before drinking sessions. Subjects received each particular dose daily until one of two criteria was met, that is, either ethanol intake did not differ by more than 20% of the mean for 3 consecutive days or for a maximum of 8 days. Both 10 and 56 mg/kg of 7-keto DHEA significantly reduced the dose of ethanol consumed. Although 10 mg/kg of 7-keto DHEA produced decreases similar to those found with DHEA, the 56-mg/kg dose of 7-keto DHEA was significantly more effective at decreasing the dose of ethanol consumed than the same dose of DHEA. These results show that 7-keto DHEA is comparable with, or possibly more effective than, DHEA at decreasing ethanol consumption in rats, and that 7-keto DHEA is a compound deserving further investigation as a possible clinical treatment for alcohol abuse without the prohormonal effects of DHEA.     

Glycyl-glutamine reduces ethanol intake at three reward sites in P rats.

beta-endorphin, implicated in modulation of ethyl alcohol reward, has neuron terminals in several reward sites. Alcohol consumption was reduced after ventricular or site-specific injections into the nucleus accumbens of an opioid-derived dipeptide, glycyl-glutamine. The current study examined the effects of this dipeptide after site-specific injections into additional reward sites. Alcohol-preferring (P) rats, stereotaxically implanted with bilateral guide cannulae into the nucleus accumbens, ventral tegmental area, and the central nucleus of the amygdala were given 30% alcohol and water in a 24h voluntary two-bottle choice paradigm. Upon achieving stable baseline intakes, glycyl-glutamine (GQ) doses were injected bilaterally, and the alcohol and water intakes and body weight recorded for the response and recovery. The data show reduced alcohol intake by 32-49.5% after 100-pmol glycyl-glutamine into reward sites (nucleus accumbens, ventral tegmental area, and central nucleus of the amygdala), but not after injections into control sites dorsal to reward sites. The order of sensitivity to the 1-fmol dose was amygdala > or = ventral tegmental area > accumbens. GQ was effective in reducing ethanol intake at reported beta-endorphin terminal regions in each of the three reward sites tested. The effective doses were similar to reported endogenous GQ levels, consistent with the notion that it may function as part of an endogenous counter regulatory mechanism and represent a "stop drinking" signal in the high drinking, P rats at these three reward sites.     

Monoamine uptake inhibitors attenuate ethanol intake in alcohol-preferring (P) rats

The P line of alcohol-preferring rats drink pharmacologically significant amounts of ethanol when given free choice between a 10 percent ethanol solution and water. Serotonin (5-HT) uptake inhibitors and desipramine, a norepinephrine (NE) uptake inhibitor, were found to significantly reduce their ethanol consumption for up to 24 hours after intraperitoneal injection. To determine if this effect of 5-HT uptake inhibitors could be altered by receptor antagonists, some of which are short acting, P rats were trained to drink ethanol by free choice during scheduled availability, with ethanol being presented one hour every four hours during the light cycle. The majority of the ethanol was consumed during the first hour of availability, and the ethanol intake was significantly reduced by the 5-HT uptake inhibitors, fluoxetine and fluvoxamine. Pretreatment with antagonists for 5-HT1, 5-HT2 and alpha- and beta-NE receptor systems failed to alter the fluvoxamine attenuation of ethanol intake. The mechanism by which 5-HT uptake inhibitors alter ethanol preference remains unclear.     

Effects of scheduled access on ethanol intake by the alcohol-preferring (P) line of rats

The effects of scheduling the availability of ethanol on its voluntary consumption by the selectively bred alcohol-preferring P rats were examined under three conditions: unrestricted 24 hr/day access (Condition A), access limited to a continuous 4 hr/day (Condition B), and access limited to 1 hr every 3 hr, 4 times/day (Condition C). Food and water were always available. Daily alcohol intakes (mean +/- SEM) with Conditions A, B and C were 6.9 +/- 0.2, 2.1 +/- 0.2 and 4.4 +/- 0.2 g/kg, respectively, while the intake per hour of availability increased from 0.3 +/- 0.03 under Condition A to 1.1 +/- 0.4 g/kg under condition C. The amount of ethanol consumed per drinking episode under Conditions A, B and C were 1.1 +/- 0.1, 2.1 +/- 0.2 and 1.1 +/- 0.03 g/kg, respectively. Mean blood alcohol concentrations (BACs), determined periodically during the dark cycle of Condition A and five minutes after drinking episodes under Conditions B and C, were 59 +/- 10, 61 +/- 7 and 62 +/- 7 mg%, respectively. When unlimited access was reinstated after Condition C, daily alcohol consumption returned to a level similar to that under the initial Condition A (7.2 +/- 0.5 g/kg). When the ethanol concentration was increased from 5 to 20% (v/v) under Condition C, the amount of ethanol consumed per episode at 5% was significantly less than at the 10, 15 and 20% concentrations, and the volume consumed was significantly lower at the 20% concentration than at the 5, 10 and 15% concentrations.(ABSTRACT TRUNCATED AT 250 WORDS)     

HAD and LAD rats respond differently to stimulating effect but not discriminative effects of ethanol.

The drug discrimination paradigm (DD) was used to evaluate behavioral differences of rats selectively bred for differential ethanol drinking preferences. Seventh-generation high alcohol-drinking (HAD) and low alcohol-drinking (LAD) rats were trained to discriminate between ethanol (0.5 g/kg, IP) and saline vehicle, following a 2-min presession interval (PI), using an FR-10 schedule of reinforcement. The HAD line was more responsive than the LAD line to the stimulating effect of ethanol as measured by total response rates. ED50 values of 0.239 and 0.244 g/kg for the HAD and LAD lines, respectively, do not reflect any difference in the discriminative effects of ethanol. Response rates during DD indicated a dissociation of rate-increasing effects and discriminative performance following ethanol. In addition to differential drinking preference, these data suggest that selective breeding for the HAD and LAD animals also involves the stimulant action of ethanol but not on the discriminative effects.     

Sugar-dependent rats show enhanced intake of unsweetened ethanol.

Rats show signs of dependence on sugar when it is available intermittently, including bingeing, withdrawal, and cross-sensitization with amphetamine. In the current study, we sought to determine whether sugar-dependent rats would show increased intake of unsweetened ethanol and, conversely, whether intermittent access to ethanol would augment sugar consumption. In Experiment 1, with intermittent versus ad libitum access to ethanol, Sprague-Dawley rats were given escalating concentrations of ethanol (1%, 2%, 4%, 7%, and 9%) over the course of 20 days. Rats in the intermittent ethanol access group, with 12-h daily access, consumed more 4%, 7%, and 9% ethanol during the first hour of access, and more 9% ethanol daily, than did rats in the ad libitum ethanol access group. In Experiment 2, with ethanol as a gateway to sugar intake, the rats from Experiment 1 were switched to 10% sucrose with 12-h daily access for 1 week. Rats in the intermittent ethanol access group consumed significantly more sugar than was consumed by rats in a control group with no prior ethanol experience. In Experiment 3, with sugar as a gateway to ethanol to determine whether sugar dependence leads to increased ethanol intake, four groups were maintained for 21 days according to the following designations: intermittent access to sugar and chow, ad libitum access to sugar and chow, intermittent access to chow, or ad libitum access to chow. Four days later, all groups were switched to intermittent ethanol access, as described in Experiment 1. The group with intermittent access to sugar and chow consumed the most 9% ethanol, supporting the suggestion that sugar dependence alters a rat's proclivity to drink ethanol. These results may relate to the co-morbidity between binge-eating disorders and alcohol intake and the tendency of people abstaining from alcohol to consume excessive amounts of sugar. In conclusion, bingeing on either ethanol or sugar fosters intake of the other.     

Soymilk products affect ethanol absorption and metabolism in rats during acute and chronic ethanol intake

In this study we evaluated the effects of soy products on ethanol metabolism during periods of acute and chronic consumption in rats. Gastric ethanol content and blood ethanol and acetaldehyde concentrations were investigated after the oral administration of ethanol (34 mmol/kg) plus soy products such as soymilk (SM) or fermented soymilk (FSM). The gastric ethanol concentration of the FSM group was greater than that of the control group, whereas portal and aortal blood ethanol concentrations of the FSM group were lower than in controls. The aortal acetaldehyde concentration in the FSM group was lower than that of the control group. The direct effect of isoflavones on liver function was investigated by using hepatocytes isolated from untreated rats. Genistein (5 micromol/L) decreased ethanol (P = 0.045) and tended to decrease acetaldehyde (P = 0.10) concentrations in the culture filtrate. Some variables of ethanol metabolism in the liver were investigated after chronic ethanol exposure for 25 d. Rats consumed a 5% ethanol fluid plus the SM diet, the FSM diet or a control diet. Microsomal ethanol oxidizing activity was significantly lower in the FSM group than the control group. Furthermore, cytosolic glutathione S-transferase activity was higher in the SM and FSM groups than in the control group. Acetaldehyde dehydrogenase activity (low K(m)) in the FSM group (P = 0.15), but not in the SM group (P = 0.31), tended to be greater than in the control group. The amount of thiobarbituric acid reacting substances in the liver of the SM and FSM groups tended to be less than that of the control group (P = 0.18 and 0.10, respectively). These results demonstrate that soymilk products inhibit ethanol absorption and enhance ethanol metabolism in rats.     

MDMA (ecstasy) substitutes for the ethanol discriminative cue in HAD but not LAD rats

Selectively bred high- and low-alcohol-drinking (HAD/LAD) rats were trained to discriminate the interoceptive stimuli produced by IP-administered 600 mg/kg ethanol (10% w/v) in a two-lever, food-motivated operant task. Once criterion discrimination was attained, animals were tested with 3.0, 1.5, 1.0, and 0.5 mg/kg MDMA. Although no differences in alcohol discrimination were observed between the HAD and LAD animals, the HAD line was significantly more sensitive than the LAD line to the effects of MDMA. These results provide additional information to the growing body of evidence suggesting serotonergic mediation of some of the behavioral effects of ethanol.     

Suppression of ethanol intake in ethanol-preferring rats by 1,4-butanediol

The oral administration of 1,4-butanediol (1,4-BD) at doses ranging from 100 to 300 mg/kg, twice daily, produced a dose-dependent reduction (40 to 85%) in the voluntary ethanol intake in rats selectively bred for high preference for ethanol. Treatment with 1,4-BD did not reduce total fluid intake. Repeated 1,4-BD administration (300 mg/kg twice daily for 7 days) suppressed ethanol intake almost completely. After suspension of 1,4-BD treatment, the inhibitory effect on ethanol intake remained significantly low for 2 days. 1,4-BD failed to inhibit aldehyde dehydrogenase to a concentration of 10 mM in rat liver homogenate.     

Hyperlipemia and early pancreatic injury induced by ethanol intake in rats

The pathogenesis of alcoholic pancreatitis is unknown, and even though hyperlipemia has been hypothesized to be a risk factor for alcoholic pancreatitis, no studies directly investigating whether there is a relationship between the two have ever been reported. Therefore, to determine if a relationship exists between hyperlipemia and alcoholic pancreatitis, especially the early stage of alcoholic pancreatic injury, we administered a regular liquid Lieber-DeCarli diet, with and without ethanol as 35% of total calories, to rats for 2 wk. Thereafter we measured their plasma lipid concentrations, pancreatic zymogen granule fragility, and plasma lipase activity and subsequently investigated the correlations between these parameters. Significant increases in plasma triglyceride, total cholesterol, phospholipid, nonesterified fatty acid, pancreatic zymogen granule fragility, and plasma lipase activity were observed in the ethanol liquid diet group, compared with the values of the control liquid diet group, and pancreatic zymogen granule fragility was correlated with plasma triglyceride (r=0.62), total cholesterol (r=0.77), phospholipid (r=0.76), nonesterified fatty acid concentrations (r=0.62), and lipase activity (r=0.63). These results show a possible relationship between hyperlipemia and the early stage of alcoholic pancreatic injury, and they may support the hypothesis that hyperlipemia contributes to the etiology of alcoholic pancreatitis.     

Selected opioids, ethanol and intake of ethanol

Rats were given an opportunity to drink tap water or a sweetened ethanol solution once a day. Across initial days of opportunity, rats increased their intake of the ethanol solution. Prior to some days' sessions with presented fluids, rats received either an injection of placebo (the carrier of drugs) or doses of ethylketocyclozocine, diprenorphine, or ethanol. Diprenorphine increased rats' intake of the ethanol solution compared to placebo. The other agents did not reliably modify intakes. These findings support a conclusion that selected activity in opioid systems of brain increase the propensity to drink alcoholic beverages.     

Experimentally induced glucose intolerance increases oral ethanol intake in rats

A number of studies have shown a relationship between glucose tolerance and ethanol intake. The present study uses a relatively simple procedure to induce glucose intolerance to test whether this condition is sufficient to produce an increase in chronic ethanol intake in male rats. Subjects were divided equally into four groups where they were given access to one of four solutions: peppermint-flavored sucrose (40%), peppermint-flavored saccharin (0.1%), peppermint in water (0.1%), or water alone presented three times a week, for a period of 11 weeks. After 12 weeks all animals were subjected to an oral glucose tolerance test which revealed that the chronically prepared sucrose animals had become glucose intolerant. At the start of week 13 all animals were given access to a 6% ethanol solution flavored with peppermint in place of the previous solutions for a period of 11 weeks. Sucrose animals displayed an immediate preference for ethanol and consumed approximately three times more ethanol than the remaining groups. The results of this study indicate that rats that are made glucose intolerant by long term access to a high concentration of sucrose, when given the opportunity, will subsequently choose to drink more ethanol than control animals.     

Selected opioids modify intake of sweetened ethanol solution among female rats

Water-deprived female rats were given a daily, 1.5-hr opportunity to take either a sweetened ethanol solution or water. Across days, they increased their intake of ethanol solution and had stable intakes of about 2 g of pure ethanol/kg after 3 weeks. Morphine (1.0 mg/kg) alone, and in combination with diprenorphine (25 micrograms/kg), increased intake of ethanol solution among females similar to the increased intake seen with males under similar procedures. Fentanyl dose-relatedly increased intake of ethanol. The data strengthen the idea that one or more of the endogenous opioid systems, but not all, are involved with instances of "excessive" intake of alcoholic beverages.     

Low doses of naltrexone reduce palatability and consumption of ethanol in outbred rats.

The opioid antagonist naltrexone, at doses of 1 and 3 mg/kg, has been shown to decrease the palatability and consumption of 10% ethanol in rats. However, a dose of 0.5 mg/kg of naltrexone has produced equivocal results. The purpose of the present study was to clarify the effects of low doses of naltrexone (0.0 [control], 0.25, 0.50, 0.75, and 1.0 mg/kg) on palatability and consumption of 10% ethanol. Sixty-four, male, Long-Evans hooded rats were divided into five groups matched for ethanol consumption. Each rat was injected over four consecutive days with one of five doses of naltrexone exactly 30 min before taste reactivity testing with 10% ethanol. When reactivity testing was completed, rats were acclimated to drink during a period of restricted access to fluid under conditions of mild fluid deprivation. Then, on four consecutive test days, rats were injected with naltrexone 10 min before 10% ethanol was made available for 30 min. Although each dose of naltrexone decreased ingestive responding to 10% ethanol over four days, this effect was not statistically reliable. However, all doses of naltrexone produced significant increases in aversive responding to the ethanol solution. Naltrexone, at the three highest doses, produced significant decreases in consumption of 10% ethanol. These results were consistent with the interpretation that naltrexone, even at low doses, reliably reduces palatability and consumption of 10% ethanol.     

Circadian activity rhythms and voluntary ethanol intake in male and female ethanol-preferring rats: Effects of long-term ethanol access

Chronic alcohol (ethanol) intake alters fundamental properties of the circadian clock. While previous studies have reported significant alterations in free-running circadian period during chronic ethanol access, these effects are typically subtle and appear to require high levels of intake. In the present study we examined the effects of long-term voluntary ethanol intake on ethanol consumption and free-running circadian period in male and female, selectively bred ethanol-preferring P and HAD2 rats. In light of previous reports that intermittent access can result in escalated ethanol intake, an initial 2-week water-only baseline was followed by either continuous or intermittent ethanol access (i.e., alternating 15-day epochs of ethanol access and ethanol deprivation) in separate groups of rats. Thus, animals were exposed to either 135 days of continuous ethanol access or to five 15-day access periods alternating with four 15-day periods of ethanol deprivation. Animals were maintained individually in running-wheel cages under continuous darkness throughout the experiment to allow monitoring of free-running activity and drinking rhythms, and 10% (v/v) ethanol and plain water were available continuously via separate drinking tubes during ethanol access. While there were no initial sex differences in ethanol drinking, ethanol preference increased progressively in male P and HAD2 rats under both continuous and intermittent-access conditions, and eventually exceeded that seen in females. Free-running period shortened during the initial ethanol-access epoch in all groups, but the persistence of this effect showed complex dependence on sex, breeding line, and ethanol-access schedule. Finally, while females of both breeding lines displayed higher levels of locomotor activity than males, there was little evidence for modulation of activity level by ethanol access. These results are consistent with previous findings that chronic ethanol intake alters free-running circadian period, and show further that the development of chronobiological tolerance to ethanol may vary by sex and genotype.     

A relation between maze performance and increased ethanol intake in Long-Evans rats.

In addition to the neurochemical and genetic basis of high ethanol consumption, there has been renewed interest in studying the role of behavioral variables and their relation to ethanol consumption. The present study was designed to assess whether a relation exists between a behavioral variable such as maze learning ability and ethanol consumption. Sixty, male, Long-Evans rats, exposed to food and water ad libitum, were given a daily trial in a complex, 16-arm T-maze for 19 consecutive days. The number of errors and time to complete the maze were monitored. Individual maze variable scores were transformed and then combined to allot each animal with an index of overall maze performance, with a smaller maze index score denoting good performance. After completion of maze testing, animals were given alternate-day, free-choice presentations of water and ascending doses of ethanol solutions (2%-10%), followed by a 10-day maintenance period with 10% ethanol. Animals were subsequently separated into two groups of high and low drinkers to examine any relation between differential ethanol intake and maze performance. A significant negative correlation between maze index and ethanol intake for the high drinkers group indicated that a smaller maze index was related to increased ethanol intake. No significant correlation was obtained for the low drinkers group. These results seem to indicate that individual variation in learning ability seems to be related to increased ethanol intake. Thus, innate learning processes may be a relevant trait when one attempts to understand the behaviors related to ethanol intake and preference.     

Predictors of ethanol consumption in adult Sprague-Dawley rats: relation to hypothalamic peptides that stimulate ethanol intake

To investigate mechanisms in outbred animals that increase the propensity to consume ethanol, it is important to identify and characterize these animals before or at early stages in their exposure to ethanol. In the present study, different measures were examined in adult Sprague-Dawley rats to determine whether they can predict long-term propensity to overconsume ethanol. Before consuming 9% ethanol with a two-bottle choice paradigm, rats were examined with the commonly used behavioral measures of novelty-induced locomotor activity and anxiety, as assessed during 15 min in an open-field activity chamber. Two additional measures, intake of a low 2% ethanol concentration or circulating triglyceride (TG) levels after a meal, were also examined with respect to their ability to predict chronic 9% ethanol consumption. The results revealed significant positive correlations across individual rats between the amount of 9% ethanol ultimately consumed and three of these different measures, with high scores for activity, 2% ethanol intake, and TGs identifying rats that consume 150% more ethanol than rats with low scores. Measurements of hypothalamic peptides that stimulate ethanol intake suggest that they contribute early to the greater ethanol consumption predicted by these high scores. Rats with high 2% ethanol intake or high TGs, two measures found to be closely related, had significantly elevated expression of enkephalin (ENK) and galanin (GAL) in the hypothalamic paraventricular nucleus (PVN) but no change in neuropeptide Y (NPY) in the arcuate nucleus (ARC). This is in contrast to rats with high activity scores, which in addition to elevated PVN ENK expression showed enhanced NPY in the ARC but no change in GAL. Elevated ENK is a common characteristic related to all three predictors of chronic ethanol intake, whereas the other peptides differentiate these predictors, with GAL enhanced with high 2% ethanol intake and TG measures but NPY related to activity.