Electrophysiological responses to ethanol, pentobarbital, and nicotine in mice genetically selected for differential sensitivity to ethanol

Cortical electroencephalographic (EEG) changes induced by ethanol (4.3 and 1.4 g/kg, ip), pentobarbital (50 and 16 mg/kg), and nicotine (1.0 g/kg) were examined in long-sleep (LS) and short-sleep (SS) mice that were genetically selected for differential sleep times induced by a hypnotic dosage of ethanol. Ethanol (4.3 g/kg) caused EEG changes that paralleled the behavioral differences, whereas no differences between selected lines were observed following the activating dose (1.4 g/kg). Data support the notion that the known difference in ethanol sleep times is due not to greater SS sensitivity to ethanol activation but rather to greater LS sensitivity to ethanol hypnosis. No differences between selected lines were observed following 50 mg/kg pentobarbital, which again parallels previous behavioral data. The SS mice were more responsive to pentobarbital activation (16 mg/kg). Nicotine more severely reduced EEG power and heart rate in LS mice; a continuous iv infusion of nicotine elicited a distinct pattern of behavioral stereotypy for each selected line, with more profound motor and reflex depression in LS mice. The lines do not differ in rate of nicotine metabolism, hence they must differ in central nervous system sensitivity to nicotine. Thus, lines of mice selectively bred for differential sensitivity to ethanol also display marked differences in electrophysiological and behavioral responses to nicotine.     

Ethanol-induced grooming in mice selectively bred for differential sensitivity to ethanol

In rats, excessive grooming follows the application of several forms of stress. The injection of alcohol may be considered as a stressor, since it increases plasma corticosterone levels. The present study examines the effects of ethanol injections on grooming in two lines of mice selected for differences in their hypnotic response to ethanol, i.e., the long sleep (LS) and short sleep (SS) lines developed at the Institute for Behavior Genetics at the University of Colorado at Boulder. Various subhypnotic doses of ethanol produced excessive grooming in the SS line, but this did not occur in the LS line. The SS mice also displayed more "novelty-induced" grooming compared to the LS mice after repeated exposure to the testing situation. The increase in excessive grooming in both the ethanol-induced and the novelty-induced excessive grooming situations was most apparent in the second half of the observation period.     

Adrenergic nervous system alteration and ethanol-induced narcosis in long-sleep and short-sleep mice

Long-sleep (LS) and short-sleep (SS) mice were pretreated with either propranolol or phentolamine, followed by a hypnotic dose of ethanol. Pretreatment with propranolol, but not phentolamine, significantly reduced ethanol sleep time in LS mice. The SS mice were not affected. In a second study propranolol pretreatment was given subsequent to ethanol at various doses, different for each line, that produced similar sleep time durations in both lines. Under these conditions, propranolol decreased sleep time in both LS and SS mice. These data lend support to the idea that noradrenergic mechanisms play a role in the mediation of the hypnotic effects of ethanol.     

Alcohol intake in selected lines of mice: importance of sex and genotype.

Mice selected by McClearn and Kakihana for differences in ethanol-induced sleep time were used as subjects. In Experiment 1, mice from the long-sleep (LS) and short-sleep (SS) lines were offered a choice of water or solution GS consisting of 3% glucose and .16% sodium saccharin (w/v); or a choice of water or solution GS + E that contained GS solution plus 4% ethanol (w/v). In Experiment 2, mice from the first experiment were provided with a three-way choice among water, solution GS, and solution GS + E. In both experiments, SS mice (alcohol-insensitive) consumed more GS + E than LS mice (alcohol-sensitive). In addition, female mice drank considerably more GS + E solution than male mice. Thus, consumption of sweetened ethanol in both a two-way choice (water and GS + E) and a three-way choice (water, GS, and GS + E) is dependent on both genotype and sex. High genetic sensitivity to ethanol was associated with low consumption, and vice versa. Although females consumed more alcohol than males, famales of these lines have not been previously found to show lower sensitivity to acute alcohol administration.     

Effects of post-training ethanol and group housing upon memory of an appetitive task in mice

It has been shown that post-training ethanol's facilitating effects upon memory disappeared if the mice were kept isolated after training. Since ethanol-treated mice were attacked by their cagemates, it has been hypothesized that the improved retention induced by ethanol resulted from an interaction between ethanol and group housing which added aversive information to training. To investigate the correctness of this interpretation, ethanol effects upon memory of an appetitive task were studied. C57BL/6J mice (isolated the day before training) were individually trained to find a cheese pellet placed in a corner of an open-field. Mice were injected intraperitoneally immediately after training with saline, 0.5, 1.5, or 2.0 g/kg of ethanol. They were then returned to their home cage and left alone, with another mouse, or with five other mice for 2 h after training. All mice were tested 24 h later for retention. Reductions in the number of pellet approaches or in the latency to eat the pellet were taken as measures of learning. Post-training ethanol disrupted retention of the appetitive task in a dose-related manner. Moreover disruption was greater in mice group housed after training. The results support the hypothesis that ethanol's post-training facilitating effects upon aversive memory may be due to added aversive information to the stimulus complex, rather than, or in addition, to enhanced storage of memory traces.     

Mice selectively bred for open-field thigmotaxis: life span and stability of the selection trait

In 2 experiments, the authors examined 69 mice selectively bred for high or low levels of open-field (OF) thigmotactic behavior (high open-field thigmotaxis [HOFT] and low open-field thigmotaxis [LOFT], respectively). They found that the strains differed in defecation during the 60-min exposure to the OF. Furthermore, the strains differed with regard to their life spans: The more thigmotactic HOFT mice lived longer than the LOFT mice. The strains were not differentiated by food intake or excretion. The strain difference in thigmotaxis was not age dependent, and it persisted in the home-cage condition as well. Neither the location (center or wall) of the starting point nor the shape (circular or square) of the OF arena affected the difference in wall-seeking behavior between the two strains. The authors concluded that the difference in thigmotaxis (or emotionality) between the HOFT and LOFT mice is a stable and robust feature of these animals.     

EFFECTS OF DRUGS AND DRUG COMBINATIONS IN PIGEONS TRAINED TO DISCRIMINAT AMONG PENTOBARBITAL,DIZOCILPINE, A COMBINATION OF THESE DRUGS,AND SALIN

Drugs with multiple actions can have complex discriminative‐stimulus properties. An approach to studying such drugs is to train subjects to discriminate among drug combinations and individual drugs in the combination so that all of the complex discriminative stimuli are present during training. In the current experiments, a four‐choice procedure was used to train pigeons to discriminate among dizocilpine (noncompetitive NMDA receptor blocker), pentobarbital (GABA_A receptor agonist), a fixed‐dose combination of these two drugs, and saline. Following extended training, low doses of pentobarbital or dizocilpine administered alone produced saline‐appropriate responding. Higher doses of pentobarbital produced responding on the pentobarbital‐appropriate key and higher doses of dizocilpine produced responding on the dizocilpine key. Administering the lowest doses of pentobarbital and dizocilpine together resulted in responding on the saline‐appropriate key. Increasing the dose of pentobarbital in the presence of low doses of dizocilpine produced responding primarily on the pentobarbital‐appropriate key; increasing the dose of dizocilpine in the presence of the lowest dose of pentobarbital produced responding primarily on the dizocilpine‐appropriate key. Combining the higher doses of pentobarbital and dizocilpine resulted in responding primarily on the drug‐combination key. Low doses of phencyclidine or ethanol produced responding on the saline‐appropriate key, but intermediate doses resulted in individual subjects responding predominately on either the pentobarbital key, the dizocilpine key, or the drug‐combination key depending on the subject. After the highest dose of phencyclidine or ethanol, most subjects responded predominantly on the drug‐combination key. Low doses of other drugs tested produced responding on the saline‐appropriate key. With the highest diazepam doses responding was largely confined to the pentobarbital‐appropriate key. The highest doses of dextromethorphan or dextrorphan resulted in responding on the dizocilpine key more frequently than on other keys. Across a range of doses, morphine produced responding predominantly on the saline key. The results using the four‐key procedure emphasized the role of both GABA_A and NMDA receptors in the complex discriminative stimulus properties of phencyclidine and of ethanol.     

Relative reinforcing effects of different oral ethanol doses in rhesus monkeys

The relative reinforcing effects of different doses of orally delivered ethanol were evaluated. Mouth-contact responding by rhesus monkeys was measured under concurrent fixed-ratio fixed-ratio schedules of liquid delivery (0.67 ml/delivery) from each of two spouts during daily 3-hr sessions. Experiment 1 examined persistence of responding with ethanol (2%, 8%, and 32% wt/vol) and water available. When fixed-ratio values from 8 to 128 were tested, the number of ethanol deliveries obtained per session decreased as the response requirement increased. The decrease in deliveries was less at higher than at lower ethanol concentrations, however. Experiment 2 examined choice between two ethanol concentrations under concurrent fixed-ratio 16 schedules (4% vs. 8%, 4% vs. 16%, 8% vs. 16%, 2% vs. 8%, 2% vs. 32%, 8% vs. 32%). Higher concentrations (16%, 32%) generally maintained more responding than concurrently available concentrations of 8% or less. An exception was the observation of a preference for 8% over 32% ethanol. When the fixed-ratio value was increased, however, the relative preference for these two doses was reversed so that 32% ethanol maintained more responding than 8% ethanol. Thus, the direction of the preference depended on the size of the response requirement. These results indicate that the reinforcing effects of ethanol increase with dose.     

Ethanol impairs memory of a simple discrimination in adolescent rats at doses that leave adult memory unaffected

Adolescent rats are less sensitive than adults to the hypothermic, anxiolytic, motor impairing, hypnotic, and lethal effects of ethanol. In vitro experiments nevertheless suggest that hippocampal neural activity is more affected by ethanol in preweanling or adolescent rats than in adults. These data are complemented by in vivo results showing that pretraining ethanol impairs learning in adolescent rats at doses that do not affect adult learning. In order to determine if posttraining ethanol affects memory differently in adolescents than in adults, Sprague-Dawley albino rats of both ages were trained in an appetitively motivated odor discrimination in which they were required to dig in scented sand for sweetened cereal reward. Immediately after training subjects received intraperitoneal injections of 0, 0.5, or 1g/kg ethanol (12.6%). At test, 48h later, subjects were presented with unbaited discriminanda and the time (s) spent digging in the S+ and S- was measured. Adolescents, but not adults, showed impaired discrimination performance if training was followed by ethanol. A subsequent experiment discounted the possibility that impaired adolescent performance was due to ethanol-induced conditioned taste or odor aversions. It thus appears that relative to adults, memory in adolescent rats is more strongly affected by ethanol in a test of appetitive conditioning that excludes ethanol's effects on sensory and motivational influences during the learning experience.     

Thyrotropin-releasing hormone (TRH) potentiates pentobarbital-based flavor aversion learning

Thyrotropin-releasing hormone (TRH), a hypothalamic polypeptide, will antagonize some of pentobarbital's major effects (sleep time and hypothermia) in rodents when administered in low doses (1-10 mg/kg). At higher doses (30 and 100 mg/kg), TRH has been shown to increase the lethality of high doses of pentobarbital in mice. The present experiments demonstrated that 10 mg/kg of TRH will potentiate the conditioned flavor aversion normally induced in rats by 20 mg/kg of pentobarbital, suggesting that the TRH-pentobarbital combination may have sublethal toxic effects even at low doses of both substances. Furthermore, this result suggests that the mechanism whereby pentobarbital produces a flavor aversion is independent of the drug's hypothermic and sleep-inducing effects.     

Drug discrimination under a concurrent fixed-ratio fixed-ratio schedule.

Pigeons were trained to discriminate 5.0 mg/kg pentobarbital from saline under a two-key concurrent fixed-ratio 10 fixed-ratio 40 schedule of food presentation, in which the fixed-ratio component with the lower response requirement was programmed to reinforce responding on one key after drug administration (pentobarbital-biased key) and on the other key after saline administration (saline-biased key). After responding stabilized, pigeons averaged 98% of their responses on the pentobarbital-biased key during training sessions preceded by pentobarbital, and they averaged 90% of their responses on the saline-biased key during training sessions preceded by saline. In test sessions preceded by doses of pentobarbital, chlordiazepoxide, or ethanol, pigeons switched from responding on the saline-biased key at low doses to responding on the pentobarbital-biased key at higher doses (the dose-response curve was quantal). High doses of phencyclidine produced responding on both keys, whereas pigeons responded almost exclusively on the saline-biased key after all doses of methamphetamine. These and previous experiments using concurrent reinforcement schedules to study drug discrimination illustrate that the schedule of reinforcement is an important determinant of the shape of dose-effect curves in drug-discrimination experiments.     

Drug discrimination under two concurrent fixed-interval fixed-interval schedules

Pigeons were trained to discriminate 5.0 mg/kg pentobarbital from saline under a two-key concurrent fixed-interval (FI) 100-s FI 200-s schedule of food presentation, and later tinder a concurrent FI 40-s FI 80-s schedule, in which the FI component with the shorter time requirement reinforced responding on one key after drug administration (pentobarbital-biased key) and on the other key after saline administration (saline-biased key). After responding stabilized under the concurrent FI 100-s FI 200-s schedule, pigeons earned an average of 66% (after pentobarbital) to 68% (after saline) of their reinforcers for responding under the FI 100-s component of the concurrent schedule. These birds made an average of 70% of their responses on both the pentobarbital-biased key after the training dose of pentobarbital and the saline-biased key after saline. After responding stabilized under the concurrent FI 40-s FI 80-s schedule, pigeons earned an average of 67% of their reinforcers for responding under the FI 40 component after both saline and the training dose of pentobarbital. These birds made an average of 75% of their responses on the pentobarbital-biased key after the training dose of pentobarbital, but only 55% of their responses on the saline-biased key after saline. In test sessions preceded by doses of pentobarbital, chlordiazepoxide, ethanol, phencyclidine, or methamphetamine, the dose-response curves were similar under these two concurrent schedules. Pentobarbital, chlordiazepoxide, and ethanol produced dose-dependent increases in responding on the pentobarbital-biased key as the doses increased. For some birds, at the highest doses of these drugs, the dose-response curve turned over. Increasing doses of phencyclidine produced increased responding on the pentobarbital-biased key in some, but not all, birds. After methamphetamine, responding was largely confined to the saline-biased key. These data show that pigeons can perform drug discriminations under concurrent schedules in which the reinforcement frequency under the schedule components differs only by a factor of two, and that when other drugs are substituted for the training drugs they produce dose-response curves similar to the curves produced by these drugs under other concurrent interval schedules.     

The effect of ethanol on stress-induced tachycardia

A study was designed to answer the questions if low doses of ethanol would reduce stress induced increases in heart rates, if covariations would be observed between ethanol induced changes in heart rates and changes in emotional states and mental performance and if tolerance to ethanol or other personality factors would influence the ethanol induced cardiac effects. Forty-four male students with a history of high and low alcohol consumption according to questionnaire scores were matched for extraversion and neuroticism and then assigned to a group receiving either 0.8 g/kg of ethanol or a placebo drink. A stress condition of mental arithmetic was applied prior to and 45 minutes after ingestion of the drink. Heart rates and ratings of emotional states by adjective check lists were recorded before and after each stress session. A significant reduction of stress induced heart rate increases in both high and low drinking groups but no ethanol dependent change of resting heart rates were observed. Reductions of autonomic stress response by ethanol were weakly but positively correlated to respective reductions of affective stress responses and impairment of the quality of mental performance. High trait anxiety subjects seemed to benefit more from ethanol with respect to reductions of cardiac and emotional arousal than low anxious subjects.     

Post-training ethanol disrupts trace conditioned fear in rats: effects of timing of ethanol, dose and trace interval duration

Ethanol has complex effects on memory performance, although hippocampus-dependent memory may be especially vulnerable to disruption by acute ethanol intoxication occurring during or shortly after a training episode. In the present experiments, the effects of post-training ethanol on delay and trace fear conditioning were examined in adolescent rats. In Experiment 1, 30-day-old Sprague-Dawley rats were given delay or trace conditioning trials in which a 10s flashing light CS was paired with a 0.5 mA shock US. For trace groups, the trace interval was 10 s. On days 31-33, animals were administered ethanol once daily (0.0 or 2.5 g/kg via intragastric intubation), and on day 34 animals were tested for CS-elicited freezing. Results showed that post-training ethanol affected the expression of trace, but had no effect on delay conditioned fear. Experiment 2 revealed that this effect was dose-dependent; doses lower than 2.5 g/kg were without effect. Experiment 3 evaluated whether proximity of ethanol to the time of training or testing was critical. Results show that ethanol administration beginning 24h after training was more detrimental to trace conditioned freezing than administration that was delayed by 48 h. Finally, in Experiment 4 animals were trained with one of three different trace intervals: 1, 3 or 10s. Results indicate that post-training administration of 2.5 g/kg ethanol disrupted trace conditioned fear in subjects trained with a 10s, but not with a 1 or 3s, trace interval. Collectively the results suggest that ethanol administration impairs post-acquisition memory processing of hippocampus-dependent trace fear conditioning.     

Effects of posttraining ethanol on an appetitive task

The present study examined the effects of posttraining ethanol administration upon retention of an appetitive task using a variety of retention behaviors associated with the task. Male C57BL/6J mice were individually trained to find a cheese pellet placed in the corner of an open field. Five behavioral measures were used including locomotor activity counts, rearings, grooming episodes, approaches to the cheese pellet, and latency to consume the cheese pellet. Immediately after training, mice were injected intraperitoneally with saline or 2.0 g/kg of ethanol and then returned to their home cage in which four "intruder" mice were added for 2 h after training. On subsequent testing days (1, 6, 14, and 51 days posttraining), mice were returned to the original training environment and the five behaviors were measured. Both saline- and ethanol-treated mice habituated to the initially novel test environment at similar rates as indicated by decreased exploratory behavior (locomotor activity and rearings). In contrast, a divergence in the latency to consume the cheese pellet was observed: Saline-treated mice behaved as though the cheese was rewarding (decreased latency to eat the pellet), while the ethanol group behaved as though the cheese was aversive (increased latency to eat the pellet). Taken with previous studies, these results demonstrate that posttraining ethanol can have strikingly different effects on retention depending on the task, the measure of retention used, and the underlying neural structures involved.     

Drug discrimination under a concurrent fixed-interval fixed-interval schedule.

Pigeons were trained to discriminate 5.0 mg/kg pentobarbital from saline under a concurrent fixed-interval (FI) FI schedule of food presentation on which, after pentobarbital administration, responses on one key were reinforced with food under an FI 60-s component and responses on the other key were reinforced under an FI 240-s component. After saline administration, the schedule contingencies on the two keys were reversed. After both pentobarbital and saline, pigeons responded more frequently on the key on which responses had been programmed to produce the reinforcer under the FI 60 component of the concurrent schedule. The schedule was changed to concurrent FI 150 FI 150 s for drug-substitution tests. In each bird, increasing doses of pentobarbital, ethanol, and chlordiazepoxide produced increases in the proportion of responses on the key on which responses had been reinforced under the FI 60 component after pentobarbital administration during training sessions. The proportion of responses on that key was slightly lower for ethanol than for chlordiazepoxide and pentobarbital. At a dose of pentobarbital higher than the training dose, responding decreased on the key that had been reinforced under the FI 60 component during training sessions. Phencyclidine produced less responding on the key programmed under the FI 60-s component than did pentobarbital. Methamphetamine produced responding primarily on the key on which responses had been reinforced under the FI 60-s component after saline administration.     

Prepartum and postpartum open-field behavior and maternal responsiveness in mice bidirectionally selected for open-field thigmotaxis

The authors examined pre- and postpartum open-field (OF) behavior and maternal responsiveness in mice that they bidirectionally selected for OF thigmotaxis. The authors tested 40 female mice under 3 conditions: prepartum OF, postpartum OF, and a pup retrieval test. In both OF conditions, the high OF thigmotaxis (HOFT) mice were more thigmotactic but explored and reared less than the low OF thigmotaxis (LOFT) mice, indicating that the HOFT mice were more emotional. In the postpartum condition, the HOFT mothers also defecated more and ambulated less than the LOFT mothers. The increase in grooming after parturition was more conspicuous among the LOFT mothers than among the HOFT mothers. The LOFT mothers were also more attracted to their pups in the OF, but the retrieval test did not show any substantial line differences. The results suggested that the line difference in emotionality was more pronounced during lactation than during pregnancy, although parturition exerted no effect on thigmotaxis.     

The effect of home or novel environment on the facilitation of passive avoidance by post-training ethanol

Passive avoidance behavior of mice is improved when mice are injected with ethanol immediately after footshock training. Further study has shown that avoidance can be affected by ethanol injections given within 1 h, but not at 90 or 180 min, after training. The present study was conducted to investigate the possibility that events which occur in the homecage during this sensitive period may influence the effect of ethanol on subsequent avoidance. Male Swiss-Webster mice were housed either singly in a novel environment for 90 min or returned to their (group) homecage following one-trial, step-through, passive avoidance training (0.1 mA footshock) and intraperitoneal injection of 3.0 g/kg ethanol (15% v/v) or saline. As in previous studies, when ethanol-treated mice were returned to their homecage, avoidance was significantly increased at 24 h compared to the behavior of saline-treated mice. However, when mice were isolated in the novel environment for 90 min immediately following treatment, the memory facilitating effects of ethanol were not observed. The avoidance behavior of mice injected with saline was the same regardless of their post-training environment. Also, the number of mice (6 or 10) housed per homecage did not significantly influence the effects of ethanol or post-training environment on avoidance. These findings indicate that environmental factors may interact with the effects of ethanol to modify avoidance behavior. The possible influence of variables such as aggression, thermoregulation, and behavioral arousal on the effects of ethanol in this paradigm are discussed.     

Drug discrimination under concurrent variable-ratio variable-ratio schedules

Pigeons were trained to discriminate 5 mg/kg pentobarbital from saline under concurrent variable-ratio (VR) VR schedules, in which responses on the pentobarbital-biased lever were reinforced under the VR schedule with the smaller response requirements when pentobarbital was given before the session, and responses on the saline-biased key were reinforced under the VR schedule with the larger response requirements. When saline was administered before the session, the reinforcement contingencies associated with the two response keys were reversed. When responding stabilized under concurrent VR 20 VR 30, concurrent VR 10 VR 40, or concurrent VR 5 VR 50 schedules, pigeons responded almost exclusively on the key on which fewer responses were required to produce the reinforcer. When other doses of pentobarbital and other drugs were substituted for the training dose, low doses of all drugs produced responding on the saline-biased key. Higher doses of pentobarbital and chlordiazepoxide produced responding only on the pentobarbital-biased key, whereas higher doses of ethanol and phencyclidine produced responding only on this key less often. d-Amphetamine produced responding primarily on the saline-biased key. When drugs generalized to pentobarbital, the shape of the generalization curve under concurrent VR VR schedules was more often graded than quantal in shape. Thus, drug discrimination can be established under concurrent VR VR schedules, but the shapes of drug-discrimination dose-response curves under concurrent VR VR schedules more closely resemble those seen under interval schedules than those seen under fixed-ratio schedules. Graded dose-response curves under concurrent VR VR schedules may relate to probability matching and difficulty in discriminating differences in reinforcement frequency.     

Effects of MK-801 and ethanol combinations on memory consolidation in CD1 mice: involvement of GABAergic mechanisms

In the present research the effect of the noncompetitive N-methyl-d-aspartate receptor antagonist MK-801 and ethanol combinations on memory consolidation and the involvement of GABAergic mechanisms in this effect were investigated in CD1 mice injected intraperitoneally with the drugs immediately or 120 min after training in a one-trial inhibitory avoidance apparatus and tested for retention 24 h later. The results showed that (a) the retention performances of mice were impaired in a dose-dependent manner by immediate posttraining MK-801 (0.2 and 0.3, but not 0.1 mg/kg) and ethanol (1 and 2, but not 0.5 g/kg) administrations; (b) an otherwise ineffective dose of MK-801 (0.1 mg/kg) enhanced the deleterious effect exerted by ethanol (1 and 2 g/kg); (c) an otherwise ineffective dose of muscimol (0.5 mg/kg) enhanced, while otherwise ineffective doses of picrotoxin (0.25 mg/kg) or bicuculline (0.1 mg/kg) antagonized, this effect; and (d) no effect was observed when the treatments were carried out 120 min after training, suggesting that the effects observed following immediate posttraining administrations were due to the influence on the consolidation of memory. From these experiments it is evident that (a) MK-801 enhances ethanol's effects on memory consolidation and (b) GABAergic mechanisms are involved in this effect.