Chronobiological regulation of alcohol intake.

Like other physiological functions, food intake and metabolism (including alcohol consumption) in humans and animal models may be regulated by circadian rhythm. For example, many studies of rodents have found that alcohol consumption in these nocturnal animals peaks during their active dark period. This alcohol consumption pattern can be influenced, however, by experimental manipulation. One factor that has been proposed to play a role in regulating circadian alcohol consumption pattern is the hormone melatonin, which is produced by the pineal gland. Research also indicates that the effects of lighting conditions on the alcohol consumption of animal models may be influenced by the differences among the strains of the laboratory animals used, variations in the type and administration schedule of the animals' alcohol-containing diet, disruptions of the normal circadian rhythm, concurrent use of other drugs, and properties of the light.

Thyrotropin releasing hormone decreases alcohol intake and preference in rats.

Thyrotropin releasing hormone (TRH) has been reported to reduce stress- and deprivation-induced eating, hypothetically by induction of satiation. Early work demonstrated thyroid extracts reduced alcohol intake, and recent research shows a TRH analog specifically inhibits alcohol preference. We determined whether parenteral administration of TRH reduces alcohol consumption and choice in a manner consistent with a satiation effect. Water-restricted ad lib fed female and male rats (n = 12) were given access to 5% w/v ethanol 0 or 30 minutes after intraperitoneal (i.p.) injection of TRH. TRH (20-40 mg/kg) inhibited alcohol intake only if injected immediately before alcohol access. Inhibition of alcohol intake was reliably accompanied by increased production of fecal boli but not by reliably decreased food intake. Rats given a choice of 2% w/v ethanol and water decreased alcohol preference after TRH (20 mg/kg) but did not reduce total fluid intake. Results are partially consistent with the hypothesis of TRH as one of several functional elements in the integrative neuropeptide control of alcohol consumption via short-term satiation.

A novel group of ovarian toxicants: the psoralens.

The psoralens are naturally occurring metabolites found in many crop plants; synthetic forms of 5-methoxypsoralen (bergapten) and 8-methoxypsoralen (xanthotoxin) are widely used in skin photochemotherapy. Our previous research documented that dietary bergapten and xanthotoxin reduced birthrates in female rats when males and females were exposed to these chemicals. The present study was designed to determine the cause of this reduced birthrate and whether this resulted from direct impact on the females. The study demonstrates that bergapten and xanthotoxin administered, either alone or in combination to female rats (mated to undosed males), significantly reduced the number of implantation sites, pups, and corpora lutea in dosed females compared with control animals. Additionally, full uterine weight and empty uterine weight were significantly reduced. These compounds also significantly reduced circulating estrogen levels in a dose-dependent manner. Interestingly, the psoralens significantly induced mRNAs of liver enzymes typically induced by polycyclic aromatic hydrocarbons, CYP1A1 and UGT1A6; the higher the dose, the greater the induction. UGT 2B1 mRNA, typically induced by phenobarbital-like compounds, was not significantly affected. Thus, enhanced oxidative metabolism and conjugation of estrogens in psoralen-treated animals may provide a partial explanation for the effects observed. These findings are also consistent with psoralen-induced reduction in ovarian follicular function and ovulation.

Interaction of CCK and 8-OH-DPAT in the satiation of alcohol intake.

Administration of the neuropeptide cholecystokinin (CCK) is known to reduce food and alcohol intake and preference. The food satiation effect of CCK is reportedly dependent on serotonergic neurotransmission. Administration of 8-OH-DPAT, a serotonin1A autoreceptor agonist, reduces the ability of CCK to inhibit feeding. We determined if CCK's alcohol satiation effect also depends on activity of serotonergic neurons by administering 8-OH-DPAT (120-240 microg/kg) to 23-h water-deprived female and male rats, followed 1 h later by i.p. injection of CCK (4 microg/kg) and 30-min access to 5% w/v ethanol. 8-OH-DPAT significantly (p < 0.05) interacted with CCK, and reduced CCK's ethanol satiation effect when given i.p. but increased CCK's effect when given s.c. Female rats showed this interaction of 8-OH-DPAT with CCK at a higher dose than males when given i.p., but females were more sensitive to s.c. 8-OH-DPAT's ability to reduce ethanol intake. Results are consistent with previous findings of dose-, sex-, and route-dependent biphasic effects of 8-OH-DPAT on feeding and ethanol intake. A partial dependence of CCK's alcohol satiation effect on serotonergic neurotransmission is revealed in this design.

Psoralen-induced growth inhibition in Wistar rats.

The linear furanocoumarins (or psoralens) are naturally occurring plant biosynthetic metabolites that have been used since ancient times in skin photochemotherapy. However, medicinal use of the psoralens has been linked with increased incidence of skin cancer in humans. To understand some of the mechanism of this toxicity, we tested increasing concentrations of the psoralens bergapten (5-methoxypsoralen) and xanthotoxin (8-methoxypsoralen) for toxicity against Wistar rats. As dietary concentrations of each compound increased, weight gain in both male and female rats decreased. Feeding on diet containing these chemicals also decreased rat birth rate, but did not significantly affect individual litter weight or date of birth. Xanthotoxin appeared to be more toxic than bergapten.

Angiotensin II reduces alcohol intake and choice in water- or food-restricted rats.

Two experiments were conducted to assess the effects of administration of the neuropeptide and hormone angiotensin II (AII) on ethanol intake and choice. First, 18 male Wistar rats were water deprived for 23 h and given access to 5% w/v ethanol for 30 min, followed by 30 min of access to water; food was ad lib. Following adaptation to this schedule, rats were randomly assigned to receive an IP injection of 0, 100, or 200 micrograms/kg of AII at either -30 or 0 min prior to ethanol access. Each AII injection decreased ethanol intake only if injected immediately before access; water and food intake were unaffected. Secondly, rats were given food daily at 2% of body weight with ad lib water and randomly assigned to receive either only water or 4% w/v ethanol ad lib on alternate days. Following adaptation, rats were randomly assigned to receive IP saline or 200 micrograms/kg of AII prior to presentation of a choice of ethanol or water for 1 h. AII reduced ethanol intake and increased water intake at 0-30 min after injection. Results confirm previous reports of inhibition of alcohol consumption by peripheral AII, and indicate a temporal constraint on AII's effect, which is consistent with a role as a short-term satiety factor.

Conditioned taste aversions induced by alcohol and lithium in rats selectively bred for ethanol neurosensitivity.

Rats that were selectively bred for differences in alcohol-induced sleep time (alcohol neurosensitivity) were tested for differences in formation and extinction of alcohol- and LiCl-induced conditioned taste aversions. Male rats bred for high, control, or low alcohol sensitivity (HAS, CAS, and LAS rats, respectively) were deprived of water and given daily 30 min access to water for a baseline period of 7 days. Rats were then given a novel 0.125% sodium saccharin solution, followed by an intraperitoneal injection of either saline, 2 g/kg of ethanol (at 10% w/v), or 50.9 mg/kg of LiCl (0.15 M) on 3 conditioning days. Each saccharin exposure was followed by a recovery day of access to water. The ethanol-induced saccharin aversion extinguished more rapidly in LAS rats than in CAS or HAS rats (p < 0.05), but LiCl conditioned equivalent aversions in each group. Also, ethanol injection results in large differences in observed resting behavior in these rats (HAS > CAS > LAS), but LiCl injection produced no reliable group differences in resting. The weaker alcohol-induced taste aversion in LAS rats accords with their previously measured higher oral consumption of alcohol (Kulkosky et al., Alcoholism 17:545-551, 1993) and the idea that alcohol intake is limited by an expectancy of postingestive consequences. The weaker ethanol-induced aversion in LAS rats reflects selective breeding of an alcohol-specific trait and not a general difference in aversive conditioning or chemical neurosensitivity.

Desmethionine-bombesin receptor antagonist blocks bombesin-induced inhibition of alcohol intake.

[D-Phe6,Des-Met14]bombesin(6-14), ethyl amide (D-BN) is a specific, competitive receptor antagonist of bombesin, a neuropeptide that inhibits alcohol and food intake. We tested the effects of IP injected D-BN (4-400 micrograms/kg) on bombesin-induced (4 micrograms/kg) reduction of caloric intake. In the first experiment, ad lib-fed female and male rats (Ns = 18) were deprived of water for 23 h, injected with peptides or saline in randomized sequences of doses, and immediately given access to 5% w/v ethanol for 30 min, followed by 30 min of water. In a second experiment, male rats (N = 10) were injected with the antagonist at 10 or 20 min prior to bombesin injection and alcohol access, and behaviors were observed and quantified once a minute with an instantaneous time-sampling technique. D-BN injection blocked the bombesin-induced reduction in alcohol intake (> or = 40 micrograms/kg) and food intake (> or = 200 micrograms/kg). When injected 20 min prior to access, D-BN alone (200 micrograms/kg) initially elevated alcohol drinking and later increased feeding behaviors and decreased resting, relative to saline injection. Results indicate bombesin-induced reduction of alcohol intake depends on a specific peptidergic receptor process, and endogenous bombesin-like peptide could act physiologically to elicit satiation with ethanol and food.

MK-329 blocks the inhibition of alcohol intake by CCK-8.

Peripheral administration of sulfated cholecystokinin octapeptide (CCK-8) potently reduces alcohol intake, preference, and blood levels in rats. MK-329 (L-364,718 or Devazepide) acts at peripheral cholecystokinin (CCKA) receptors to antagonize CCK-8's physiological and behavioral effects, such as pancreatic stimulation and inhibition of feeding. We determined whether CCKA receptor blockade would also prevent CCK-8's alcohol satiety effect. Water-deprived female and male rats (n = 7 for each) received randomized combinations of intraperitoneal injections of MK-329 (0, 100, 200, or 400 micrograms/kg) followed by CCK-8 (0 or 4 micrograms/kg). Rats were then given access to 5% w/v ethanol for 30 min, followed by 30-min access to water, with food ad lib. MK-329 at all doses significantly (p < 0.05) reduced the suppression of alcohol intake and food intake by CCK-8. MK-329 alone increased alcohol intake at 400 micrograms/kg, and increased food intake, in females and males at 100 and 200 micrograms/kg, respectively. We concluded that CCK-8's alcohol and food satiation effects depend on specific, peripheral CCKA receptors, and satiation of alcohol consumption and drinking-associated feeding reflect an endogenous functional interaction of CCK-8 with CCKA receptors.

Cholecystokinin and bombesin inhibit ethanol and food intake in rats selectively bred for ethanol sensitivity.

Cholecystokinin octapeptide (CCK-8) and bombesin tetradecapeptide (BBS-14) are brain-gut neuropeptides shown to inhibit intake and choice of alcohol solutions and foods in a variety of species. Recently, Draski and colleagues selectively bred strains descended from N/Nih outbred Norway rats that differ in sleep time after injection of ethanol. The intake of 5% w/v ethanol, food, and water was measured in these rats with high, low, and control alcohol sensitivity (HAS, LAS, and CAS), after intraperitoneal injection of randomized sequences of doses of CCK-8 or BBS-14 (0-8 micrograms/kg). During baseline adaptation to water deprivation-induced consumption of alcohol, LAS rats consumed reliably more ethanol than HAS or CAS rats. Injection of CCK-8 or BBS-14 significantly and equivalently suppressed intake of ethanol and food at 30 min after presentation in each group of rats. Water intake and food intake at 30-60 min following alcohol access was not affected by prior injection of either neuropeptide. Large differences in alcohol neurosensitivity (HAS > CAS > LAS) were observed in these rats' resting behavior for 1 hr after intraperitoneal injection of 1 g/kg of ethanol. These selectively bred alcohol neurosensitivity differences cannot be explained by corresponding differences in sensitivity to the inhibitory behavioral effects of CCK-8 or BBS-14. However, differences in alcohol intake and resting behavior do correspond to artificially selected sensitivities to ethanol's hypnotic effect.

Bombesin reduces alcohol choice in nutritive expectancy and limited-access procedures.

Bombesin is a bioactive tetradecapeptide found in nerves of the brain and gut and previously shown to inhibit intake of ethanol in forced-choice, one-bottle tests in water-deprived rats. In the present experiments, intraperitoneal bombesin (4-100 micrograms/kg) reduced selection of alcohol in two-bottle choice tests with water. In an application of the "nutritive expectancy" procedure, weight-reduced rats received access to 4% w/v ethanol and water. Bombesin injection suppressed the intake of ethanol, but not of water, in rats with prior ethanol experience. In an application of the "limited access" procedure, nondeprived rats received access to 6% w/v ethanol and water in nonhome cages during either the light or the dark phase of a 12:12 h lighting cycle. Bombesin injection lowered the intake of ethanol, but not of water, in both phases of the lighting cycle and in both sexes. Water deprivation is not necessary for bombesin to inhibit alcohol intake and this effect cannot be explained by hypodipsia, nonspecific debilitation, or conditioned aversion. Endogenous bombesin-like neuropeptides may specifically reduce choice of alcohol by signaling satiation with ethanol.

Effects of peripheral neurotensin on behavior of the rat.

Neurotensin (NT), a tridecapeptide found in brain and gut neurons, inhibits feeding and grooming, increases drinking, and enhances ethanol-induced sedation in rats after central injection. We tested the behavioral effects of IP injection of NT (0.1-100 micrograms/kg) in water-deprived rats given access to 5 or 10% ethanol for 30 min, followed by 30-min access to water. Behaviors during alcohol access were quantified with an instantaneous time-sampling observational technique. Food intake and observed feeding and grooming behaviors were significantly inhibited by large doses of NT (10-100 micrograms/kg) and water intake and resting behavior were increased. When the "limited access procedure" was used to induce ethanol selection in nondeprived rats, NT did not affect ethanol or water intake. Peripheral NT affects intake of food and water and observed feeding, grooming, and resting after peripheral injection in deprived rats, but does not affect ethanol consumption. These actions suggest physiological roles for endogenous neurotensin and its receptors in regulation of specific behaviors.

Effects of cholecystokinin on morphine-elicited hyperactivity in hamsters.

The effects of the octapeptide cholecystokinin (CCK) on hamster locomotor activity were investigated in three experiments. In Experiment 1, the effect of CCK (25, 50, 75 micrograms/kg) on morphine (2.5 mg/kg)-elicited hyperactivity was studied. Results indicated that CCK antagonized morphine-elicited hyperactivity and that CCK alone elicited hypoactivity. There were no effects of dose of CCK. In Experiment 2, the effects of intraperitoneal (IP) and subcutaneous (SC) routes of administration of CCK (25 micrograms/kg) on locomotor activity were studied. Compared to saline controls, CCK induced hypoactivity that was of greater magnitude and of longer duration when administered IP than SC. Experiment 3 was designed to replicate the route of administration effect observed in Experiment 2 and to determine whether sensitization to CCK-induced hypoactivity develops over the course of a few injections. Results indicated that CCK-induced hypoactivity was greater after IP than SC administration but that sensitization was not detectable. It is concluded that CCK antagonizes morphine-elicited hyperactivity in the hamster by acting, in part, independently of morphine to produce opposite behavioral effects.

Cholecystokinin-induced satiation with ethanol: effects of lighting cycle and limited access procedures.

Cholecystokinin (CCK) is a brain-gut neuropeptide and hormone previously shown to inhibit alcohol intake in water- or food-deprived rats. The effects of CCK and the phase of lighting cycle on alcohol intake in rats were investigated in a comparison of limited access and water-restriction procedures. The limited access procedure (LAP) is a recently developed technique for inducing free-choice alcohol consumption in nondeprived animals. Two groups of 12 male rats each were maintained in either normal or reversed 12:12 L:D lighting cycles and simultaneously given 40 minutes' access to 6% w/v ethanol and water in nonhome cages. After adaptation to this procedure, CCK octapeptide (0.5-16 micrograms/kg) was injected IP prior to access to fluids. During LAP, CCK reduced alcohol intake and increased water intake more potently in the dark phase. These effects of CCK were more reliable when the design was replicated, which suggests the importance of acquired expectancies for the development of CCK's actions. CCK more effectively reduced alcohol intake in LAP, than in a 23.3-h water-deprivation procedure for inducing alcohol intake in a 2-bottle choice test with water. However, CCK was less so effective in LAP, than in the water-deprivation procedure when alcohol was presented alone in a 1-bottle test. The alcohol satiation effect of CCK is independent of prior deprivation and not an artifact of thirst reduction, debilitation, or conditioned aversion, because CCK strongly increased water intake in the limited access procedure, and ethanol preference remained robust after experience with CCK. CCK may operate endogenously as a specific factor in satiation with ethanol.

Effects of caffeine and bombesin on ethanol and food intake.

The methylxanthine caffeine and ethyl alcohol are widely used and powerful psychotropic drugs, but their interactions are not well understood. Bombesin is a brain-gut neuropeptide which is thought to function as a neurochemical factor in the inhibitory control of voluntary alcohol ingestion. We assessed the effects of combinations of intraperitoneal (i.p.) doses of caffeine (CAF, 0.1-50 mg/kg) and bombesin (BBS, 1-10 micrograms/kg) on 5% w/v ethanol solution and food intake in deprived rats. Deprived male and female Wistar rats received access to 5% ethanol or Purina chow for 30 minutes after i.p. injections. In single doses, CAF and BBS significantly decreased both ethanol and food consumption, at 50 mg/kg and 10 micrograms/kg, respectively. CAF and BBS combinations produced infra-additive, or less-than-expected inhibitory effects on ethanol intake, but simple additive inhibitory effects on food intake. This experimental evidence suggests a reciprocal blocking of effects of CAF and BBS on ethanol intake but not food intake. Caffeine, when interacting with bombesin, increases alcohol consumption beyond expected values. Caffeine could affect the operation of endogenous satiety signals for alcohol consumption.

Vasoactive intestinal peptide: behavioral effects in the rat and hamster.

The behavioral effects of intracerebroventricular (ICV) injection of the brain-gut peptide vasoactive intestinal peptide (VIP) were quantified with a behavioral sampling technique in home-caged, nondeprived, male and female albino rats and golden hamsters. ICV VIP sex-dependently decreased observed resting behavior during 1 hr after injections in both rats and hamsters at 0.1-10.0 micrograms. Grooming behavior was increased in hamsters, and rearing and standing behaviors were increased in rats, sex-dependently at VIP doses that decreased resting. Drinking behavior was suppressed in rats by VIP at 10.0 micrograms. Intraperitoneal (IP) VIP (100.0 micrograms/kg) increased 5% ethanol intake and decreased eating behavior in fluid-deprived male rats. The increase in ethanol intake produced by IP VIP was prevented by IP cholecystokinin octapeptide (CCK, 4.0 micrograms/kg). VIP potently controls resting and ingestive behaviors, suggesting a role for this neuropeptide, along with CCK, in the feedback regulation of rodent behavior.

Cholecystokinin and satiation with alcohol.

Release of the brain-gut peptide cholecystokinin (CCK) is stimulated by intragastric instillation of ethanol, and peripheral administration of CCK inhibits ethanol consumption. To assess the temporal specificity of the inhibitory effect of CCK on alcohol intake, water-deprived rats were given 5% ethanol at 20, 10 or 0 min after intraperitoneal injections of CCK octapeptide. Delaying access to ethanol for 20 min prevented a significant effect of CCK on intake. CCK's temporally constrained inhibitory action on alcohol consumption is consistent with an ethanol satiation effect. To test the motivational specificity of CCK's effect on fluid intake, rats were allowed a 2-bottle choice of 2% ethanol and water after CCK injections. Ethanol solution intake was suppressed by CCK, and total water intake was unaffected. The putative alcohol satiation action of CCK is appropriately specific to ethanol solution in free-choice tests. Hungry, but not fluid-deprived rats that were either ethanol experienced or naive received a 2-bottle choice of 4% ethanol or water after CCK or saline injections. CCK again specifically inhibited ethanol intake, but this effect required prior ethanol experience. Doses of CCK and naloxone, an opioid receptor blocker, combined to inhibit ethanol intake in an infra-dose-additive manner in water-deprived rats. CCK may act endogenously, in part on opioid receptor-mediated processes, as a preabsorptive satiety signal of ethanol. The full expression of this action appears to depend on prior conditioning of nutritive expectancy of the postingestive effects of alcohol.

Alpha-theta brainwave training and beta-endorphin levels in alcoholics.

An alpha-theta brainwave biofeedfack training program was applied as a novel treatment technique for chronic alcoholics. Following a temperature biofeedback pretraining phase, experimental subjects completed 15 30-min sessions of alpha-theta biofeedback training. Compared to a nonalcoholic control group and a traditionally treated alcoholic control group, alcoholics receiving brainwave training (BWT) showed significant increases in percentages of EEG record in alpha and theta rhythms, and increased alpha rhythm amplitudes. Alcoholics receiving BWT showed a gradual increase in alpha and theta brain rhythms across the 15 experimental sessions. These experimentally treated alcoholics showed sharp reductions in self-assessed depression (Beck's Depression Inventory) compared to the control groups. Alcoholics receiving standard medical treatment (abstinence, group psychotherapy, antidepressants) showed a significant elevation in serum beta-endorphin levels at the conclusion of the experiment. This neuropeptide is an index of stress and a stimulant of caloric (e.g., ethanol) intake. Application of brainwave treatment, a relaxation therapy, appears to counteract the increase in circulating beta-endorphin levels seen in the control group of alcoholics. 13-month follow-up data indicate sustained prevention of relapse in alcoholics that completed alpha-theta brainwave training.

Neuropeptide Y: behavioral effects in the golden hamster.

Neuropeptide Y (NPY) is found abundantly in nervous tissues of vertebrate species including the golden hamster. Centrally-administered NPY has been reported to elicit ingestive behaviors in the rat, squirrel, pig, mouse, and chick. To assess NPY's behavioral effects in a New World rodent that does not increase food intake after deprivation, NPY was injected intracerebroventricularly (10.0-0.04 micrograms/5 microliter) in home-caged golden hamsters with ad lib access to food, water and 5% w/v ethanol solution. Food and fluid intakes, and behavior displays were monitored after NPY injection. NPY promptly increased short-term food intake and observed feeding behaviors at 10.0, 3.3, 1.1, and 0.37 micrograms NPY, but there was no effect on 24 hr food intake. Water and ethanol intakes were increased only at 10.0 and 0.37 micrograms NPY, respectively. Resting behaviors decreased at NPY doses that increased feeding, but there were no consistent effects of NPY on any other category of behavior. Results demonstrate that NPY potently stimulates short-term food intake and decreases resting behavior in the golden hamster. The lack of compensatory food intake in deprived hamsters cannot be explained as an insensitivity to the putative orexigenic function of endogenous neuropeptide Y.

Effect of bombesin on behaviors associated with ethanol satiation and blood ethanol levels.

The behavioral specificity and physiological significance of bombesin-induced inhibition of ethanol intake were assessed in water-deprived rats. The behavioral display accompanying suppression of 5% ethanol intake by bombesin tetradecapeptide (BBS-14, 1-4 micrograms/kg) was measured with an instantaneous time-sampling technique. Blood ethanol levels were measured after peripheral BBS-14 and bombesin nonapeptide (BBS-9) administration, and after either oral self-administration or peripheral injection of ethanol. The display accompanying BBS-14-reduced ethanol consumption differed from control in that less drinking and feeding behaviors were observed and resting increased, dose-dependently. The typical behavioral sequence of ethanol satiation was observed in all conditions. Both BBS-14 and -9 reduced blood ethanol levels when oral intake was suppressed, and BBS-14 did not affect blood ethanol levels or elimination rate when ethanol was injected. The results are compatible with an hypothesis of a functional role for endogenous bombesin-like peptides and receptors in a neuropeptide control of ethanol intake and energy balance.