Sensitivity to low doses of ethanol and pentobarbital in mice selected for sensitivity to hypnotic doses of ethanol.

These experiments assessed sensitivity to low doses of ethanol and pentobarbital in mice that had been selectively bred with respect to ethanol sleep time (the length of time an animal remains on its back following a hypnotic dose of ethanol). The hypothesis under investigation was that short-sleep (SS) mice might be more sensitive than long-sleep (LS) mice to excitatory effects produced by low doses of depressants. In support of this hypothesis SS mice were more active in an open-field test after ethanol than were LS mice. The lines did not differ in performance on a rotating-rod apparatus after these same doses of ethanol, suggesting that the difference in open-field activity was not attributable to a greater impairment of locomotor activity in LS mice. A similar difference in the open-field activity of the selected lines was observed with pentobarbital.     

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.     

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.     

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.     

Arousal as an explanation for differences in rats selectively bred for differential alcohol sensitivity

A common error made by behavior genetics researchers is breeding two lines for differences in central arousal rather than for a specific behavioral feature. Two lines of rats (Riley, Freed, & Lester, 1976) were selectively bred for locomotor impairment in response to a subhypnotic dose (1.5 g/kg) of ethanol. These lines (designated "most affected" and "least affected") were compared in a variety of tests and showed differences in a number of phenotypic traits in addition to locomotor impairment to ethanol. The published findings have been interpreted in light of a hypothesis suggesting adventitiously selected differences in central arousal between the two lines. This interpretation showed that their usefulness as animal models of alcoholism is seriously compromised.     

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.     

Behavioral,subjective, and electroencephalographic indices of sleep onset latency and sleep duration

Electrographic (EEG, EOG, EMG) indices have been used for some decades in the definition of the stages of sleep and more recently in the diagnosis of sleep-related disorders, e.g., insomnia, despite the lack of detailed information concerning the precise relationship between such electrographic indices and behavioral and subjective criteria of sleep. Evaluation of the relationship between EEG, behavioral, and self-report measures of sleep onset latency and sleep duration was conducted using 17 young normal sleepers. The behavioral measure was provided by an apparatus which records subjects' button-press responses to an auditory stimulus presented at various interstimulus intervals (2, 5, and random 1, 2, 5, 7, and 10 min). The behavioral and stage 2 EEG estimates of sleep onset latency (SOL) and sleep duration (SD) were almost identical. The stage 1 EEG provided the shortest estimate of SOL; the self-report measure, the longest. The SD measures were in reverse order. There were no significant differences among the three interstimulus interval conditions (2, 5, and random min). Average response rates to the 50-dB chime were 100% during wakefulness, 81% during EEG stage 1, and 8% during EEG stage 2. Almost all stage 2 responding occurred during the first 5 min of each stage 2 period. The validity of electrographic indices as sleep criteria and the implications of the findings for the formulation of an adequate definition of sleep and its clinical measurement are discussed.     

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.     

Differential sensitivity to the effects of nicotine and bupropion in adolescent and adult male OF1 mice during social interaction tests

Few studies have compared the action of both nicotine (NIC) and bupropion (BUP), an antidepressant used to treat NIC dependence, on social and aggressive behavior at different ages. This study aims to determine whether these drugs produce differential effects in adolescent (postnatal day: 36-37) and adult (postnatal day: 65-66) mice that have been housed individually for 2 weeks in order to induce aggressive behavior. Mice received BUP (40, 20, or 10 mg/kg), NIC (1, 0.5, and 0.25 mg/kg as base), or vehicle earlier to a social interaction test. BUP (40 mg/kg) decreased social investigation and increased nonsocial exploration in both adolescent and adult mice. The same effects were also observed in adult mice administered with a lower dose of the same drug (20 mg/kg). In adolescents, NIC (1 mg/kg) decreased social investigation, but this effect did not reach statistical significance in adults. In conclusion, a differential sensitivity to the effects of NIC or BUP emerged in some of the behavioral categories when the two age groups were compared.     

Ethanol state-dependent memory: involvement of dorsal hippocampal muscarinic and nicotinic receptors

In the present study, the effects of bilateral injections of cholinergic agents into the hippocampal CA1 regions (intra-CA1) on ethanol state-dependent memory were examined in mice. A single-trial step-down passive avoidance task was used for the assessment of memory retention in adult male NMRI mice. Pre-training intraperitoneal injection (i.p.) of ethanol (0.25, 0.5 and 1 g/kg) dose dependently induced impairment of memory retention. Pre-test administration of ethanol (0.5 and 1 g/kg, i.p.) induced state-dependent retrieval of the memory acquired under pre-training ethanol (1 g/kg, i.p.) influence. Pre-test intra-CA1 injection of physostigmine (2.5 and 5 μg/mouse, intra-CA1) or nicotine (0.3 and 0.5 μg/mouse, intra-CA1) improved pre-training ethanol (1 g/kg)-induced retrieval impairment. Moreover, pre-test administration of physostigmine (2.5 and 5 μg/mouse, intra-CA1) or nicotine (0.3 and 0.5 μg/mouse, intra-CA1) with an ineffective dose of ethanol (0.25 g/kg) significantly restored the retrieval and induced ethanol state-dependent memory. Pre-test intra-CA1 injection of the muscarinic receptor antagonist, atropine (4 and 8 μg/mouse, intra-CA1) or the nicotinic receptor antagonist, mecamylamine (2 and 4 μg/mouse, intra-CA1) 5 min before the administration of ethanol (1 g/kg, i.p.) dose dependently inhibited ethanol state-dependent memory. Pre-test intra-CA1 administration of physostigmine (0.5, 2.5 and 5 μg/mouse), atropine (2, 4 and 8 μg/mouse), nicotine (0.1, 0.3 and 0.5 μg/mouse) or mecamylamine (1, 2 and 4 μg/mouse) alone cannot affect memory retention. These findings implicate the involvement of a dorsal hippocampal cholinergic mechanism in ethanol state-dependent memory and also it can be concluded that there may be a cross-state dependency between ethanol and acetylcholine.     

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.     

Effects of peripheral immune activation on social behavior and adrenocortical activity in aggressive mice: Genotype-environment interactions

To explore genetic-developmental differences in the biobehavioral effects of induced illness, males from two lines of mice selectively bred for high or low levels of aggressive behavior were injected with endotoxin (Escherichia coli, LPS: 0.25 mg/kg, 1.25 mg/kg, or 2.5 mg, i.p.) or saline. Body temperature, weight, and locomotor activity were monitored immediately before and 8 and 24 hr after injection. Twenty-four hours after injection, social behaviors were assessed in a 10-min dyadic test, and hypothalamus, spleen, and serum were collected. In both lines, endotoxin treatment increased behavioral immobility ("freezing") and decreased social exploration. Other effects showed line differences: Males from the high-aggressive line had a lower threshold to endotoxin-induced effects on body temperature, weight loss, spleen weight, and corticosterone. Social reactivity (startle response to mild social investigation) increased in the high-aggressive line and decreased in the low-aggressive line after treatment. In the high-aggressive line only, endotoxin decreased attack frequency and increased latency to attack. The interactions between selected line (genotype) and endotoxin treatment (environment) demonstrate that genetic-developmental differences in social and aggressive behavior may indicate the extent to which immune stimuli (e.g., bacteria, viruses, cytokines) function as "biobehavioral stressors." Aggr. Behav. 23:93–105, 1997.© 1997 Wiley-Liss, Inc.     

Chlordiazepoxide effects on ethanol self-administration: dependence on concurrent conditions.

Experiments examined the effects of acute doses of chlordiazepoxide upon ethanol self-administration in the rat. A concurrent-schedule procedure was used that employed choice between ethanol (5%) and a second fluid (either water or a 1% sucrose solution). When ethanol and water were the available fluids, chlordiazepoxide at doses of 15 and 20 mg/kg reduced ethanol-reinforced responding and intake, with a greater reduction occurring at the 20 mg/kg dose. However, when ethanol and sucrose were concurrently available, in many rats only the 20 mg/kg dose of chlordiazepoxide reduced ethanol-reinforced responding. The differences in dose response function occurred in most animals without large changes in the baseline ethanol-reinforced responding across the two concurrent conditions. Thus the dose-effect curve relating chlordiazepoxide and ethanol self-administration can be altered, dependent upon the nature of the concurrently available reinforcers.     

Environmental familiarization in rats: differential effects of acute and chronic nicotine

If an environment is familiar, rats will interact more with a novel object than if the environment is unfamiliar. In two experiments we used this behavioral tendency to assess the effects of nicotine on environmental familiarization (i.e., an elevated platform). As expected, rats given 2 min of exposure to the platform on 2 consecutive days (familiarization phase) interacted more with a novel object in a subsequent test than rats that had not experienced the platform until the test day. During the familiarization phase acute pretreatment with nicotine (0.6 and 1.8 mg/kg, subcutaneous) 10 min before platform exposure interfered with familiarization processes, as measured by object interaction on the drug-free test day. Behavioral measures of activity (e.g., turning and midline crosses) eliminated an account based on nicotine-induced motor impairment. Furthermore, this effect of acute nicotine on familiarization was not due to nonspecific effects of nicotine. Controls that received equivalent nicotine exposure temporally separated from platform exposure interacted more with the novel object than similarly treated controls that were unfamiliar with the platform on the test day. Interestingly, rats treated once daily with 0.6 mg/kg nicotine for 14 days before the familiarization phase (chronic condition) did not show a decrease in environmental familiarity. This dissociation extends a growing literature finding that the behavioral and neurobiological effects of nicotine differ, in part, after acute and chronic exposure. Indeed, acute nicotine (0. 2, 0.6, and 1.2 mg/kg) in the present report consistently decreased the amount of time spent with one paw on the edge of the platform; chronic nicotine did not affect this behavior.     

Developmental effects of acute, chronic, and withdrawal from chronic nicotine on fear conditioning

Pre-adolescence and adolescence are developmental periods associated with increased vulnerability for tobacco addiction, and exposure to tobacco during these periods may lead to long-lasting changes in behavioral and neuronal plasticity. The present study examined the short- and long-term effects of nicotine and nicotine withdrawal on fear conditioning in pre-adolescent, adolescent, and adult mice, and potential underlying substrates that may mediate the developmental effects of nicotine, such as changes in nicotinic acetylcholine receptor (nAChR) binding, CREB expression, and nicotine metabolism. Age-related differences existed in sensitivity to the effects of acute nicotine, chronic nicotine and nicotine withdrawal on contextual fear conditioning (no changes in cued fear conditioning were seen); younger mice were more sensitive to the acute effects and less sensitive to the effects of nicotine withdrawal 24 h post treatment cessation. Developmental differences in nAChR binding were associated with the effects of nicotine withdrawal on contextual learning. Developmental differences in nicotine metabolism and CREB expression were also observed, but were not related to the effects of nicotine withdrawal on contextual learning 24 h post treatment. Chronic nicotine exposure during pre-adolescence or adolescence, however, produced long-lasting impairments in contextual learning that were observed during adulthood, whereas adult chronic nicotine exposure did not. These developmental effects could be related to changes in CREB. Overall, there is a developmental shift in the effects of nicotine on hippocampus-dependent learning and developmental exposure to nicotine results in adult cognitive deficits; these changes in cognition may play an important role in the development and maintenance of nicotine addiction.     

Sex differences in adolescent ethanol drinking to behavioral intoxication

Rodent models have been especially useful for investigating adolescent ethanol exposure. However, there is a paucity of studies examining sex differences in behavioral intoxication from adolescent ethanol drinking. Here, we used an ethanol drinking model to investigate if adolescent rats of both sexes readily drink ethanol to measurable behavioral intoxication, indicated by increased impulsive action and motor incoordination. Beginning on postnatal day (P) 28, male and female Long‐Evans rats were given 30‐min access to a solution of sucrose (20%) or sweetened ethanol (20% sucrose +15% ethanol) every other day until P60 and once after 2 weeks of forced abstinence (on P75). On alternate (nondrinking) days, rats were reinforced with a food pellet for making a cued nosepoke response. Beginning on P56, rats were tested in this task after drinking sessions to assess ethanol‐induced changes in impulsive action, defined as premature responding prior to cue presentation. Motor coordination was assessed before and after drinking sessions using an incline plane test. Adolescent male and female rats readily consumed ethanol to behavioral intoxication, measured as reduced motor coordination. Following forced abstinence, females displayed greater ethanol‐induced impulsive action. These studies provide evidence for sex differences in behavioral intoxication following adolescent ethanol drinking.     

The effects of concurrent manipulations of cholinergic and noradrenergic systems on neocortical EEG and spatial learning

In the spatial learning test, young animals were divided into three groups receiving saline, scopolamine (0.15 mg/kg), or scopolamine (0.8 mg/kg). Half of the animals in each group were lesioned with DSP-4 to destroy noradrenergic fibers. DSP-4 lesions did not produce any significant impairment alone or in combination with a lower dose of scopolamine (0.15 mg/kg), but they did further augment the scopolamine (0.8 mg/kg)-induced defect. In the electroencephalography (EEG) experiment, both control rats and DSP-4-lesioned rats were recorded after receiving saline, scopolamine (0.15 mg/kg), and scopolamine (0.8 mg/kg) injections. Scopolamine induced a dose- and behavioral state-dependent EEG slowing, whereas DSP-4 lesions did not change either baseline EEG activity or EEG reactivity to scopolamine.     

Nicotine vs. ethanol discrimination: Extinction and spontaneous recovery of responding

Studies regarding extinction and spontaneous recovery of the discriminative stimulus effects of drugs are limited. Eight rats were initially trained to discriminate nicotine (0.4 mg/kg) vs. ethanol (800 mg/kg). For four rats, itraperitaneal (IP) administrations of nicotine fifteen minutes prior to fifteen-minute training sessions served as a discriminative stimulus (S^D) for predicting food-reinforced lever pressing (VI-1 min). On other sessions ethanol functioned in predicting nonreinforcement (S^Δ). The stimulus roles of the drugs were counterbalanced for the remaining four rats. S^Δ and S^D sessions alternated quasi-randomly with two daily sessions at 1000 and 1400 hours. Discriminative control was not disrupted following ten extinction sessions under a non-drug/saline condition, but was disrupted following extinction sessions under the original training drugs. Instances of spontaneous recovery (SR) occurred throughout extinction under the drug condtions. There was no evidence for SR two weeks following extinction, but partial recovery four weeks following the final extinction phase. Contextual status (context renewal) had neither a restorative or disruptive impact on extiguished or discriminated responding, respectively. Theser results support and extend the limited number of other studies by demonstrating extinction and spontaneous recovery of responding discriminated bytwo distinct drugs. Some theoretical interpretations regarding history effects and training in the context of drug discrimination are entertained.     

Characterization of a dose-response curve for nicotine-induced conditioned taste aversion in rats: relationship to elevation of plasma beta-endorphin concentration

In the first experiment a conditioned taste aversion paradigm was used to characterize a dose-response curve for the aversive properties of nicotine in male Sprague-Dawley rats. Doses of nicotine ranging from 0.01 to 0.46 mg/kg, 2.0 ml of 0.47 M lithium chloride, or saline were injected, ip, 10 min after exposure to a novel saccharin solution. Amount of saccharin consumed in a two-bottle test was assessed 72 h later. Nicotine doses of 0.046 mg/kg and above produced a significant degree of conditioned taste aversion. In a second experiment, four groups of 10 rats each were injected with saline, 0.022 mg/kg nicotine, 0.46 mg/kg nicotine, or 2.0 ml 0.47 of M LiCl. Doses of 0.46 mg/kg nicotine and 0.47 M LiCl elevated plasma beta-endorphin concentrations significantly above saline control values. The 0.022 mg/kg dose, the highest dose that did not produce conditioned taste aversion in Experiment 1, did not significantly increase plasma beta-endorphin concentrations. This finding suggests that doses of nicotine that produce conditioned taste aversion also promote the release of pituitary stress hormones. Taken together these data suggest that some of the pharmacological and behavioral effects attributed to nicotine, including the release of endogenous neuromodulators, may be dose-dependent concomitants of the aversive effects of nicotine in nicotine-naive animals.     

Effects of moderate chronic ethanol consumption on hippocampal nicotinic receptors and associative learning

A number of studies have reported that ethanol exposure induces changes in different brain systems. The hippocampus is a brain region that is very vulnerable to ethanol exposition, which functionally results in impairment of learning and memory processes reported in heavy drinkers. Hippocampal nicotinic receptors are involved in learning and memory. In this study, we determined the effects of ethanol on the main hippocampal subtypes of neural nicotinic receptors (α7 and α4β2) in rats non-selected for alcohol consumption, in order to check for possible changes on these receptors that could be linked with alterations in learning acquisition. Binding assays were carried out with [³H]methyllycaconitine ([³H]MLA) to study the α7 and [³H]nicotine to study α4β2 receptors. Auto-shaping, continuous ratio and extinction procedures were used as behavioral tests. The results show that moderate chronic ethanol consumption for 10 weeks produces: (a) a decrease of both hippocampal nicotinic receptor subtypes without alterations in affinity; (b) no differences in behavioral performance between control rats and ethanol-drinking rats in auto-shaping and continuous ratio; (c) an improvement of performance of extinction paradigm. These results indicate that chronic ethanol consumption, at moderate levels, induces changes in hippocampal nicotinic receptors but does not impair acquisition and performance of new associative learning and even improves some kind of paradigms. These results may have implications in the biochemical basis of interactions between alcohol and nicotine and the effects of these drugs on behavior.