Effect of neuropeptide Y microinjected into the hypothalamus on ethanol consumption.

Guide cannula were implanted in rats aimed at the paraventricular nucleus (PVN) of the hypothalamus for microinjection of neuropeptide Y (NPY), D-NPY27-36, or vehicle. In the Wistar rat, there was no significant effect on the consumption of ethanol. In Myers' high ethanol preferring (mHEP) rats, D-NPY27-36 caused a significant 54% decrease in ethanol consumption from baseline, but the response was not different from vehicle. NPY-induced feeding in satiated Wistar rats, was blocked by a Y1 receptor antagonist, D-NPY27-3). D-NPY27-36 decreased 78% feeding in food-deprived rats. Thus, neither the Wistar nor the mHEP rat perceives ethanol as a source of calories comparable to food.

Differences in brain area concentrations of dopamine and serotonin in Myers' High Ethanol Preferring (mHEP) and outbred rats.

Both male and female mHEP rats consume excessive amounts of ethanol and thus offer a rational model for examining biochemical and behavioral differences with non-drinking rat lines. Differences in basal concentrations of 5-hydroxytryptamine (5-HT) and dopamine (DA) correlate with the consumption of ethanol in some ethanol-preferring rat lines. The concentrations of 5-HT and DA were examined by HPLC in five brain areas (prefrontal cortex, hippocampus, nucleus accumbens, striatum and hypothalamus) of ethanol-näive rats and after the oral administration of 0.25 or 1.0 g ethanol/kg in the male and female mHEP rat, the male Wistar rat, and the female Sprague-Dawley rat. The mHEP and control rats that received ethanol were screened for drinking in a 10-day "step-up" 3% to 30% ethanol solutions beginning at postnatal days 40 and 80, and then tested at 150 days of age. The levels of DOPAC in females were lower in the hippocampus of both naïve mHEP and ethanol-treated Sprague-Dawley rats. In striatum, the concentrations of 5-HT and DA were elevated in both mHEP and ethanol-treated Sprague-Dawley female rats. The concentrations of 5-HT and its metabolite, 5-HIAA, were lower in the nucleus accumbens of the ethanol-näive female mHEP rat relative to the female outbred control. In the male rats, the levels of DA, HVA and DOPAC, as well as 5-HT and 5-HIAA were reduced in the hypothalamus of both ethanol-näive mHEP rats and Wistar rats receiving ethanol by gavage. These data demonstrate differences in neurotransmitter activity between the selectively bred mHEP rat and the outbred rat strains. There are few common features found in both the male and the female mHEP rat when compared to their respective controls. Differences in neurotransmitter function in these brain areas may account for some of the behavioral differences previously demonstrated between the two sexes of the mHEP rat.

Tiospirone and the reinforcing effects of cocaine in the conditioned place preference paradigm in rats.

Tiospirone (TSP) is an atypical antipsychotic drug. It has 5HT-2 antagonistic properties as well as affinity for D2, 5HT-1a, 5HT-6 and sigma receptors. Behavioural studies in our laboratory, which used a 24h free access to food and fluids paradigm, showed a decreased alcohol and increased food intake after twice-daily administration of TSP; the maximal effect was obtained at a dose of 0.48 mg kg(-1). This study used the conditioned place preference paradigm to determine the effect of TSP on the reinforcing properties of cocaine. Intraperitoneal administration of 5.0 mg kg(-1) cocaine, but not saline, increased the time rats spent in the drug-paired compartment of a three-compartment shuttle box by 104.9%. Two doses of TSP, 0.143 and 0.48 mgkg(-1), were tested subcutaneously 60 min before saline or cocaine administration during the conditioning phase only. A dose-response effect was observed with a significant reduction in the time rats spent in the cocaine-paired compartment on the drug-free test day produced by the dose of 0.48 mg kg(-1) (an increase of only 38.1% when post-conditioned times were compared with preconditioned times). These findings suggest that TSP reduces the reinforcing properties of cocaine exhibited in the conditioned place preference paradigm.

Effects of pregnancy and progesterone on the consumption of ethanol by the high ethanol preferring (HEP) rat.

A significant fraction of women continue to drink heavily during pregnancy, which is associated with the fetal alcohol syndrome, alcohol-related birth defects, alcohol-related neurodevelopmental disorder, and spontaneous abortion. The objective of this study was to determine whether the selectively bred genetic drinking Myers High Ethanol Preferring (HEP) rat would continue to drink through pregnancy. Rats from the F7 generation were screened by a 10-day 3-30% (v/v) ethanol concentration 'step up' procedure in order to determine the concentration which resulted in maximal drinking with an ethanol solution to total fluid ratio closest to 0.5. After baseline drinking of the preferred concentrations was established, female HEP rats were randomly selected for mating and their ethanol bottles were removed. Upon finding a 'sperm plug', male rats were removed and the ethanol was returned. A second group received injections of progesterone in sesame oil beginning with a 1.0 mg/kg/day dose which was increased to 3.0 mg/kg/day on gravid days (GD) 5-20. Vaginal smears confirmed that progesterone rendered the rats anoestrous. Neither pregnancy nor progesterone changed either the amount or proportion of ethanol consumed compared to the baseline period. The rats drank an average of 8.4 g/kg daily throughout pregnancy. A sharp drop in food intake was noted the day after mating. Beginning on GD 13, it was observed that pregnant rats showed a marked increase in the variance for proportion of ethanol consumed and body weight. Subsequently, only one of the eight impregnated rats successfully delivered a litter. The ethanol solution was removed and these rats mated again: seven of the eight rats delivered litters. These two findings suggest that the pregnant females must have begun to lose their litters on or after GD 13. Further, pregnancy does not affect the consumption of ethanol in the HEP rat. In addition, due to the fact that drinking by HEP rats during pregnancy leads to such a high rate of resorption of the fetus, this hybrid strain may also constitute a useful model for the study of alcohol-induced spontaneous abortion.

The cardiovascular effects of amperozide: interactions with cocaine.

Amperozide is a 5-HT2A receptor antagonist that significantly reduces the acquisition and expression, by rats, of a cocaine conditioned place preference. In order to rule out the possibility that amperozide affects a cocaine conditioned place preference due to effects on blood pressure or heart rate, the cardiovascular effects of amperozide were investigated. Alternating cumulative doses of amperozide (0.5, 1.0, 1.5 and 2.5 mg kg(-1)) or saline and phenylephrine (8 microg kg(-1)) were administered through the femoral vein of awake freely-moving Sprague-Dawley rats and blood pressure and heart rate were recorded from the femoral artery. A single dose of cocaine (5.0 mg kg(-1)) was administered after all the amperozide or saline doses were given. Amperozide (0.5, 1.0, 1.5 and 2.5 mg kg(-1)) did not have any significant effect on blood pressure compared to the saline control treatment to the same animals. However, 0.5 mg kg(-1) amperozide significantly decreased heart rate at 5 and 10 min. after administration. but higher doses did not further depress heart rate. Amperozide did not affect the increased blood pressure and decreased heart rate caused by phenylephrine. an alpha1-adrenoceptor agonist. In addition, amperozide did not affect the cardiovascular response to an intravenous dose of 5.0 mg kg(-1) cocaine. These results suggest that amperozide does not cause direct cardiovascular effects. The mechanism by which the lowest dose of amperozide caused a decrease in heart rate is unknown. Amperozide affects neither alpha-adrenoceptor mediated vasoconstriction nor the increased sympathetic activity caused by the peripheral and central effects of cocaine. The significance of these results, in terms of locomotor activity and the cocaine conditioned place preference paradigm, is discussed.

Effects of amperozide and tiospirone, atypical antipsychotic 5-HT2 drugs, on food-reinforced behavior in rats.

Amperozide (AMPZ) is a 5-HT2 receptor antagonist that can decrease consumption of ethanol and prevent cocaine conditioning of a place preference. Tiospirone (TSP) is a 5-HT2 receptor antagonist with affinity for D2, 5-HT1a, and 5-HT7, and sigma receptors, which can decrease consumption of ethanol while increasing food intake. Both drugs were tested for inhibition of food reinforced bar-pressing behavior. Fasted Sprague-Dawley male rats were trained daily on a 15-min FR-5 schedule. After response rates stabilized, each rat in one group received a 60-min pretreatment s.c. with saline, 0.25,0.75, or 2.5 microlmol/kg of AMPZ (injections were twice per week and counterbalanced). Each rat in a second group received a 60-min pretreatment with saline, 0.1,0.3, or 1.0 micromol/kg of TSP. The dose of 2.5 micromol/kg of AMPZ reduced the number of food reinforcements by 21% from saline. The effect was due to a greater decline in bar pressing during the last 5-min block. TSP, 0.3 and 1.0 micromol/kg, significantly reduced the number of reinforcements by 45 and 94% from saline, respectively. In a study of catalepsy, TSP at 0.3 and 1.0 micromol/kg produced akinesia and catalepsy, respectively. Our results indicate that AMPZ-induced reduction of alcohol intake is not due to a nonspecific effect on reward, but the akinetic effects of TSP masks whether or not it has effects on reward.

Genetics of alcoholism: rapid development of a new high-ethanol-preferring (HEP) strain of female and male rats.

A genetically based animal model of alcoholism has been developed in a relatively short period of 3 years. The new strain is characterized by an intense preference for ethanol over water as well as unique behavioral, neurochemical and other attributes. This new strain, termed high-ethanol-preferring (HEP) rats, was derived initially from selective cross-breeding of a variant strain of female Harlan Sprague-Dawley (SD) rats with the outbred Wistar line of male ethanol-preferring (P) rats. In this study, drinking patterns of both genders were obtained over 10 days by presenting water and ethanol in concentrations ranging from 3% to 30%. To expedite the development of the new strain, only three to five female and male rats served as breeders, which were chosen from all litters on the basis of their maximum g/kg intake integrated with proportion of ethanol to total fluid values. Profiles of intake of preferred concentrations of ethanol were obtained over 24 h of unlimited access as well as during 2-h intervals of limited access to ethanol. Levels of blood ethanol were measured in both female and male HEP animals during bouts of ethanol drinking in the limited access paradigm. By the sixth generation of HEP rats, ethanol consumption of the females often exceeded that of any other rat genetically bred to drink ethanol (e.g., at a concentration of 15.7%, 10.3 g/kg per day). Seven additional characteristics are notable: 1) the HEP rats prefer ethanol in the presence of a nutritious chocolate drink or nonnutrient sweetened solution (aspartame); 2) high levels of blood ethanol are associated with their drinking; 3) females drink significantly greater g/kg amounts of ethanol than HEP males and prefer a higher percent concentration of ethanol; 4) the drinking of ethanol by the female HEP animals does not fluctuate during the estrous cycle; 5) neurochemical assays show differential profiles of 5-HT, dopamine, and their metabolites in different regions of the brain; 6) measures of activity using the elevated plus maze, open field, and cork gnawing reveal differences between genders of HEP rats and SD rats; and 7) the HEP animals are without phenotypically expressed abnormalities. Finally, one cardinal principle derived from this study revealed that the breeding strategy to develop high-ethanol-drinking rats centers on the use of multiple solutions of ethanol whereby the intakes of ethanol in concentration of 9% through 20% dictate the ultimate selection of breeding pairs over successive F generations. Further, it is concluded that because of an intense rise in ethanol drinking of the F1 generation of female HEP rats well above that of the parental SD female breeders, the complex genotypic characteristic of the male P rat is predominantly responsible for evoking ethanol drinking in female offspring.

Role of taste and calories in the selection of ethanol by C57BL/6NHsd and Hsd:ICR mice.

The C57BL/6 mouse (C57) is used as a model for the human consumption of ethanol. Previous studies on the taste preferences of the C57 mouse indicate that ethanol drinking by this animal is for calories and not for a pharmacological effect. The purpose of this study, therefore, was to further determine the role of calories and taste in the selection of ethanol by the C57 mouse. C57 and outbred Hsd:ICR (ICR or CD-1) mice were housed two per cage with three drinking tubes. A standard 10-day preference test of 3-30% ethanol (v/v) vs. water was performed: the mean maximally preferred concentrations of ethanol were 17.9% for C57 and 6.8% for ICR mice. Once drinking of the preferred concentration for each cage had stabilized at 13.2 and 0.9 g/kg/day, respectively, the third tube was filled with water, 0.5% aspartame, isocaloric dextrose, or diluted chocolate Ultra Slim-Fast plus dextrose. Five days of dextrose or chocolate drink reduced the amount of ethanol consumed by 41% and 44% by C57 mice, but aspartame did not affect their drinking. Additional groups of C57 and ICR mice were habituated to a 2-h limited access to water. When offered a 0.5 mM quinine solution as the only fluid, both strains consumed the same volumes as water. Presentation of a saccharin solution was followed by an i.p. injection of either 0.5 M LiCl or NaCl. When given the saccharin solution 48 h later, the LiCl-treated mice of both strains drank less saccharin. The C57 mouse did not exhibit a LiCl-induced taste aversion when ethanol was the novel solution. As a test of response to novelty, a cork stopper was placed in each cage. The ICR mice gnawed much more of the cork than did the C57 mice. Thus, both C57 and ICR mice learned a taste aversion, but the C57 mouse altered its large consumption of ethanol based on more palatable sources of calories. These data support the earlier concept that the consumption of ethanol represents a preferred source of calories for the C57 mouse. Extrapolation of genetic or biochemical differences between these mice to differences between the human alcoholic and the nonalcoholic should thus be made with caution.

Comparison of the alcohol-preferring P rat to the Wistar rat in behavioral tests of impulsivity and anxiety.

To determine whether the selectively bred alcohol preferring P rat displays impulsive and anxiety-related behaviors, as have been noted in Type 2 human alcoholics, P rats were compared with outbred Wistar rats, the strain from which P rats were derived, on a series of behaviors reflecting impulsivity and anxiety. The two groups were also compared on their volitional consumption of ethanol. When compared with the Wistar rats, the P rats preferred a higher concentration of ethanol and imbibed a much greater amount of ethanol when they were offered their preferred concentration. However, the behavioral tasks produced inconsistent results. The P rats completed 100 bar presses for food in less time when tested on a constant reinforcement schedule, which suggests that they are hyperactive compared to Wistar rats. However, the P rats also emitted a higher percentage of reinforced responses on differential reinforcement of low rate responding (DRL)-10s and gnawed less from a cork stopper, which suggests that they are less impulsive and possibly neophobic. The two groups did not differ on emergence into or activity in an open field, their activity in or open-arm duration in the elevated plus maze, or performance on DRL-5s and DRL-15s. Collectively, the behavioral data suggest that P rat does not serve as a model for the anxiety and impulsiveness associated with the Type 2 alcoholic individual.

Volitional consumption of ethanol by fawn-hooded rats: effects of alternative solutions and drug treatments.

Behavioral and neurochemical measures of brain 5-hydroxytryptamine (5-HT) function in the Fawn-Hooded rat are abnormal relative to outbred strains of rats. Fawn-Hooded rats freely drink large amounts of 10% ethanol in the presence of water and have been proposed to be an animal model for studies related to alcoholism. In this study, Fawn-Hooded rats were given solutions of ethanol increasing in concentration from 3% to 30% (w/v in tap water) over 10 days with tap water in a second drinking tube and a third tube left empty. The solutions of ethanol that produced maximal drinking with a preference (ml ethanol/ml total fluid) near 50% ranged from 5% to 13%, which became the fixed individual concentrations for each rat. After a 5-day baseline period the rats were offered a solution in the third drinking tube of either 0.5% aspartame or chocolate Ultra SlimFast (diluted with water 2:1). The chocolate drink, but not aspartame, significantly reduced the consumption of alcohol by 73%. For the drug experiments, the rats were given successive 4-day periods of: baseline drinking; drug or saline injections b.i.d.; and a posttreatment period. Neither ipsapirone, a 5-HT1a partial agonist, nor naltrexone injected inhibited the intakes of ethanol solutions. Treatment with 2.5 mg/kg of amperozide, a 5-HT2 antagonist, decreased the consumption of ethanol by 38%, but also caused a decrease in consumption of food. These results show a pattern of drinking of increasing concentrations of ethanol different than other strains of rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Comparison of the action of the 5-HT2 antagonists amperozide and trazodone on preference for alcohol in rats.

Previous studies in the rat demonstrated that the 5-hydroxytryptamine2 (5-HT2) antagonist amperozide attenuates the volitional intake of both alcohol and cocaine solutions in a free-choice situation. However, another 5-HT2 antagonist, ritanserin, has not been found to reduce alcohol drinking consistently in the rat. In this study, trazodone was compared to amperozide for its effect on the volitional consumption of alcohol because, like amperozide, trazodone is a potent 5-HT2 receptor antagonist but a weak inhibitor of 5-HT reuptake. Male Sprague-Dawley rats were induced to drink alcohol by 10 mg/kg cyanamide injected for 3 days b.i.d. One week later the rats were offered a choice of water and increasing concentrations of alcohol solutions ranging from 3% to 30% v/v in a three-bottle two-choice paradigm. After the concentration of alcohol that produced maximal daily intake was determined for each rat, trazodone or amperozide was injected b.i.d. SC in doses of 1.0 mg/kg or 2.5 mg/kg for three days. Whereas the higher dose of amperozide produced a significant, 55.6% decrease from pretreatment baseline of alcohol intake, trazodone did not alter alcohol preference at either the 1.0- or 2.5-mg/kg dose. These results are discussed in terms of whether the antagonism of 5-HT2 receptors by amperozide is critical to its attenuating effect on preference for alcohol solutions.

Prenatal exposure to cocaine: effects on aggression in Sprague-Dawley rats.

Social/aggressive behavior in adult rat offspring (beginning at postnatal Day 180) prenatally exposed to saline, cocaine, or amfonelic acid (AFA) was examined. Pregnant rats received injections of 15 mg/kg of cocaine, or 0.9% saline twice daily, s.c., or on 2 consecutive days at 4-day intervals, or 1.5 mg/kg amfonelic acid daily throughout gestational Days 1-20. Frequency, duration, and latency of 11 social/aggressive behaviors were recorded for two 15-min sessions during which a smaller male intruder replaced an ovariectomized female in the resident's home cage. Subjects received a s.c. saline injection before Session 1 and 2.0 mg/kg of gepirone, a 5HT1a partial agonist, prior to Session 2. Prenatal cocaine treatment resulted in alterations of aggressive behavior. Aggressive behavior was reduced by gepirone in all groups but to a lesser extent in the AFA group.

Changes in dopamine, serotonin and their metabolites in discrete brain areas of rat offspring after in utero exposure to cocaine or related drugs.

The concentrations of dopamine (DA), serotonin (5HT) and their metabolites were quantified in 5 brain areas of rats exposed to saline, cocaine (15 mg/kg b.i.d.), amitriptyline (10 mg/kg), or amfonelic acid (AFA, 1.5 mg/kg) throughout gestation. Male pups from 3 similarly treated dams were fostered to 2 surrogate dams. The process of breeding and rearing was repeated 4 times with new dams to build the groups to 4-12, since only one pup per litter was used for any one measurement. AFA was used to mimic the dopamine (DA) uptake blockade and stimulant properties of cocaine and amitriptyline was used to mimic the other pharmacological effects of cocaine. At postnatal days (PND) 30, 60, and 180, one pup per litter was removed for HPLC analysis of monoamines. A second pup received 0.3 mg/kg haloperidol, catalepsy assessed after 1 hr, and the brain used for analysis. The cataleptic response to haloperidol was unaffected by any prenatal treatment. The striatum from PND 30 cocaine rats had decreased levels of DA without a decrease in DA metabolites. At PND 60 in cocaine exposed rats, DA and DOPAC concentrations were increased, and 5HT levels were decreased in the striatum. The amitriptyline-exposed group exhibited decreased 5HT and 5-HIAA levels in the striatum. The hypothalamus of the cocaine group had lower levels of 5-HIAA, and other brain areas had a trend for lower levels of 5HT and 5-HIAA. At PND 180, DOPAC was increased in the striatum and prefrontal cortex of the cocaine group. Haloperidol-induced altered monoamine metabolism was unaffected by any prenatal treatment at any age. These data suggest that age-related changes in the DA and 5HT neurotransmission systems occur in rats exposed prenatally to cocaine. However, the ability of the dopaminergic system to respond to a challenge by a DA receptor blocker is unaltered by these in utero treatments.

Amperozide, a 5-HT2 antagonist, attenuates craving for cocaine by rats.

Amperozide, a novel 5-HT2 receptor antagonist with little affinity for the dopamine receptor, suppresses the intake of alcohol in rats without affecting food intake or inducing other side effects. Because of these actions, amperozide was examined for its efficacy on the oral preference by the rat for a solution of cocaine. In this study, rats were selected for their voluntary consumption of at least 10 mg/kg of cocaine per day in a two-choice paradigm. A solution of 0.02% to 0.06% cocaine plus 0.03% saccharin in water was offered to each animal simultaneously with a solution of only 0.03% saccharin in water. The consumption of food and both fluids, as well as body weight, was recorded daily for three successive periods: 4 days of pretreatment baseline; 3 days during injections of either amperozide or the saline vehicle solution; and 4 days postinjections. Amperozide was administered SC twice daily in a dose of 0.5, 1.0, or 2.5 mg/kg. The volitional intake of cocaine was significantly reduced not only during the 3-day period of injections of amperozide but also during the 4-day posttreatment period. Amperozide exerted little or no effect on the intake of food or on body weight. Radioligand binding experiments confirmed that amperozide has at least a twentyfold greater affinity for 5-HT2 receptors in the frontal cortex of the rat, as compared to striatal DA1 and DA2 receptors, with the proportion value similar to that of the 5-HT2 receptor antagonist, ritanserin.(ABSTRACT TRUNCATED AT 250 WORDS)

Prenatal exposure to cocaine. I: Effects on gestation, development, and activity in Sprague-Dawley rats.

Sperm-positive Sprague-Dawley rats received one of four treatments for 20 days beginning within 24 hours of conception. One group received subcutaneous injections of 15 mg/kg cocaine twice daily (Cocaine-D); a second group received 15 mg/kg cocaine twice daily for two consecutive days at 5-day intervals (Cocaine-I); a third group received normal saline twice daily (Saline); and a fourth group received 1.5 mg/kg amfonelic acid (AFA), a dopamine reuptake inhibitor, once daily. Cocaine-D, Cocaine-I, and AFA dams were fed ad lib. An attempt was made to pair-feed the Saline dams with the Cocaine-D dams; however, the Saline dams did not eat as much as the Cocaine-D dams which resulted in dams in all groups essentially eating ad lib. The Cocaine-D pups showed a slightly delayed righting behavior and neophobia at 30 days of age, as evidenced by hypoactivity during the first 15 min of a 6-h activity test. The Cocaine-I pups were hypoactive during the 3-h dark phase of the 6-h activity test when tested at 30 days of age. These effects did not occur in the offspring exposed to AFA, a potent dopamine uptake inhibitor and CNS stimulant which indicate that one or more other sites for cocaine action may combine for its effects on the developing fetus.