Chronic intermittent injections of high-dose ethanol during adolescence produce metabolic, hypnotic, and cognitive tolerance in rats

BACKGROUND: Many humans are first exposed to ethanol during adolescence, the time at which they are most likely to binge drink ethanol. Chronic intermittent ethanol (CIE) exposure produces ethanol tolerance in adolescent rodents. Recent studies suggested that adolescent animals administered CIE experienced increased cognitive impairment following an ethanol challenge. These studies further explore development of ethanol tolerance caused by CIE in adolescence, and whether CIE during adolescence leads to altered ethanol response in adulthood. METHODS: Beginning postnatal day (P) 30, adolescent rats were administered 5.0 g/kg ethanol or saline every 48 hours for 20 days. In experiment I, animals were tested for differential weight gain. In experiment II, loss of righting reflex (LORR) was observed after each injection, then at completion of pretreatment all animals were tested with 5.0 g/kg ethanol and LORR was observed. In experiment III, blood ethanol levels were observed and elimination rates calculated after the first and fifth pretreatments. All animals were tested with 5.0 g/kg at completion of pretreatment and elimination rates were recalculated. In experiment IV, animals were trained on the spatial version of the Morris Water Maze Task (MWMT) on non-treatment days. Following completion of pretreatment and training, animals were tested after receiving an ethanol (1.0, 1.5, or 2.0 g/kg), or saline. Tests for experiments II, III, and IV were repeated in the same animals following 12 ethanol-free days. RESULTS: Chronic intermittent ethanol exposure during adolescence caused differential weight gain (experiment I). Adolescent rats developed tolerance to ethanol-induced LORR (experiment II) and metabolic tolerance to ethanol (experiment III). This tolerance was seen after 12 ethanol-free days. CIE also attenuated ethanol-induced spatial memory deficits in the MWMT (experiment IV). This effect was not long-lasting. CONCLUSIONS: Following CIE pretreatment during adolescence, tolerance developed to the hypnotic and cognitive impairing effects of ethanol, along with increased metabolic rate and decreased weight gain. These results further emphasize the ability of CIE to produce a variety of effects during adolescence, some having long-lasting consequences.     

Age differences in the expression of acute and chronic tolerance to ethanol in male and female rats

Background: Ontogenetic differences in response to ethanol (EtOH) challenge have been observed under a variety of circumstances, including varying reports of developmental differences in the expression of tolerance to EtOH. The purpose of the present experiment was to further explore potential differences in acute (AT) and chronic (CT) tolerance expression between adolescent and adult, male and female Sprague–Dawley rats, using the social interaction test. Methods: AT and CT to the social suppressing effects of a moderate dose of EtOH was assessed in adolescent and adult rats following intraperitoneal injections of 2.0 g/kg EtOH or saline daily for 10 days. At test, adults and adolescents were challenged with 1.0 or 1.25 g/kg EtOH, respectively, with AT and CT assessed at 5 and 25 minutes postinjection using ratios of impairment to brain ethanol concentrations (BrECs) at each time period (CT) and within‐session declines in impairment relative to BrECs (AT). Results: In adolescents, 10 days of EtOH pre‐exposure resulted in evidence of CT at 25 minutes postinjection, perhaps associated with an enhanced expression of AT. Among adults, signs of CT were seen at 5 minutes postinjection in adults, and may reflect neuroadaptations unassociated with AT, as with evidence of tolerance emerging only in adult control animals repeatedly exposed to saline injection prior to EtOH challenge on test day. Sex differences in tolerance expression were not observed at either age. Conclusions: Our results show ontogenetic differences between adolescents and adults in the short‐ and long‐term neuroadaptations that they express in response to repeated perturbations with EtOH. Together these findings add age of exposure and time of testing within the intoxication period as critical variables to be considered when exploring the complex relationship between AT and CT.     

Ethanol intake in the juvenile, adolescent, and adult rat: effects of age and prior exposure to ethanol

Background: Initial ingestion of ethanol by naïve rats has seemed to decrease dramatically with age. During the preweanling period, infant rats consume large quantities of high concentrations of ethanol without initiating procedures, in some instances exceeding doses required for severe motor incoordination. During adulthood, however, initial ingestion of ethanol without initiation procedures is low and infrequent. In the present study, the ontogeny of ethanol intake was measured in juvenile, adolescent and adult rats using a technique [consume off the floor (COF)] similar to that used to study intake during infancy. How this initial experience with ethanol affected subsequent affinity for ethanol intake was later assessed using 2‐bottle choice preference tests. Methods: Independent ingestion of ethanol was measured at 3 developmental periods, the juvenile period (P22–P28), adolescence (P30–P34) and adulthood (P60–P64), with systematic variation in ethanol concentration (15 or 30% v/v) and palatability (sweetness) of ethanol. Blood ethanol concentrations (BECs) were determined in all animals. This dependent variable served as an estimate of absolute ethanol ingestion. Three COF sessions were conducted for each age group. Following these sessions animals' ethanol consumption was also assessed using a 2‐bottle choice test (water vs 15% v/v unsweetened ethanol). Results: In all experiments, groups consuming 30% v/v ethanol exhibited significantly higher BECs than those exposed to 15% v/v ethanol. Adding saccharin to the ethanol increased absolute ethanol ingestion in only the oldest animals. During the pre‐exposure phase (COF sessions) of each experiment, absolute ethanol intake was found to decline with repeated exposures. Sex effects were particularly evident during later stages of ontogeny (adolescents and adults). The overall pattern of results indicated that juveniles relative to adults show a marked predisposition to consume highly concentrated ethanol solutions and that BECs derived from the COF sessions influenced ethanol acceptance patterns in the subsequent 2‐bottle test. Conclusions: Using the (COF) technique with BECs as an estimate of intake, absolute ethanol consumption seems to be quite high early in ontogeny and decline gradually into adulthood. Adding saccharin to ethanol solutions at the concentration used in the present study (0.1%) was generally not sufficient to increase absolute ethanol intake from the floor, except during adulthood. The experimental strategy employed in this study represents a novel approach for examining ethanol acceptance patterns across ontogeny and how experience with the process of intoxication affects subsequent ethanol preferences.     

Effect of the selective NMDA NR2B antagonist, ifenprodil, on acute tolerance to ethanol-induced motor impairment in adolescent and adult rats

Background: Adolescent rats have been observed to be less sensitive than adults to a number of acute ethanol effects, including ethanol‐induced motor impairment. These adolescent insensitivities may be related in part to the more rapid emergence of within session (acute) tolerance in adolescents than adults. Adolescent‐related alterations in neural systems that serve as ethanol target sites, including changes in NMDA receptor subunit expression, may influence the responsiveness of adolescents to acute ethanol effects. This study explored the role of NMDA NR2B receptors in the development of acute tolerance to ethanol‐induced motor impairment in male adolescent [postnatal day (P)28–30] and adult (P68–70) Sprague–Dawley rats. Methods: Motor‐impairing effects of ethanol on the stationary inclined plane and blood ethanol concentrations (BECs) were examined following challenge at each age with a functionally equivalent ethanol dose (adolescents: 2.25 g/kg; adults: 1.5 g/kg). Data were collected at two postinjection intervals (10 or 60 minutes) to compare rate of recovery from ethanol intoxication with BEC declines using the Radlow approach ( Radlow, 1994 ) and changes in motor impairment/BEC ratios over time for assessing acute tolerance. Results: Both vehicle‐treated adolescent and adult animals showed similar acute tolerance development to the motor‐impairing effects of ethanol at these functionally equivalent doses on the stationary inclined plane, as indexed by an increasing time‐dependent dissociation between BECs and ethanol‐induced motor impairment, with motor impairment declining faster than BECs, as well as by significant declines in motor impairment/BEC ratios over time. Acute tolerance development was reliably blocked by administration of the NR2B antagonist, ifenprodil, (5.0 mg/kg), in adult rats, whereas adolescents were affected by a higher dose (10.0 mg/kg). Conclusions: These data support the suggestion that alterations in NMDA receptor systems occurring during adolescence may contribute to reduced sensitivity to ethanol by enhancing the expression of acute tolerance development in adolescents relative to adults.     

The effects of NMDA and GABAA pharmacological manipulations on acute and rapid tolerance to ethanol during ontogeny

BACKGROUND: Sensitivity to several ethanol effects increases during ontogeny, perhaps in part because of a notable decline in acute tolerance. In contrast, rapid tolerance to ethanol-induced sedation emerges slowly during ontogeny. This study tested the hypothesis that ontogenetic differences in glutamate and/or gamma-aminobutyric acid systems influence tolerance expression. METHODS: Sprague-Dawley rats at postnatal day (P)26 or P70 received (+)MK-801, muscimol, or saline before ethanol (3.5 or 4.5 g/kg) or saline on day 1 and ethanol only on day 2. Loss of and time to regain the righting reflex and blood alcohol levels at recovery were recorded. The presence of acute tolerance was indicated as a positive slope of the linear regression of blood alcohol levels at recovery versus ethanol dose. Rapid tolerance was estimated on day 2 by comparing animals given ethanol only on day 2 with those given ethanol on both days. RESULTS: Acute tolerance on day 1 only was observed at P26; this was disrupted by (+)MK-801 but not muscimol. Evidence for acute tolerance also emerged in adults on day 2. Whereas both drugs increased ethanol sedation at both ages, they did not facilitate ontogenetic expression of rapid tolerance: rapid tolerance was not evident at P26 regardless of pretreatment when indexed in terms of recovery time. CONCLUSIONS: These data provide further evidence for an ontogenetic dissociation in the expression of acute and rapid tolerance to ethanol-induced sedation. Pharmacological attenuation of the expression of acute tolerance was sufficient but not necessary to delay recovery of righting after ethanol. The greater propensity of young animals to develop acute tolerance, seemingly modulated in part by NMDA receptors, may contribute to their relative resistance to ethanol, although other factors, including pharmacokinetic factors, also contribute to their more rapid recovery from ethanol sedation.     

Sex differences in ethanol-induced hypnosis and hypothermia in young Long-Evans rats

Introduction: Females experience greater liver damage, have reduced brain size, and have greater memory deficits than do males with a similar history of alcoholism. Females have higher peak alcohol levels and faster elimination rates than males. Our goal was to study sex differences in the response of young ethanol‐naïve outbred Long‐Evans rats to acute ethanol exposure so that we may better understand why females are more sensitive to alcohol toxicity than males. Methods: Females aged 49 days and males aged 43 days, weighing 153.6 and 177.5 g, respectively, were tested for their initial response to ethanol. Fasted (12 hr) females (in diestrous) and males were given an intraperitoneal injection of 3.0 g/kg of ethanol (v/v in 0.9% sterile saline). Body temperature, loss of the righting reflex (LORR), return of the righting reflex, and tail blood alcohol concentration (BAC) were monitored. Results: LORR occurred at the same time in females and males. The return of the righting reflex occurred later in males than in females. BACs were the same in the males and females except at LORR, when BAC was lower in the males. Acute ethanol tolerance developed in more males than females. Females demonstrated a slower recovery from peak ethanol‐induced hypothermia than males. The proportions of lean body mass, ethanol elimination, and ethanol metabolism were similar in the females and males. Conclusions: Ethanol‐naïve young male and female Long‐Evans rats demonstrated sex differences in their initial responses to ethanol. Males were more sensitive than females to the hypnotic effect of ethanol, whereas females were more sensitive than males to ethanol‐induced hypothermia. In addition, more males than females developed acute ethanol tolerance. Investigating the mechanisms underlying these differences may help us to understand why females experience more of the adverse effects of alcohol consumption than males.     

Sensitivity and tolerance to autonomic effects of ethanol in adolescent and adult rats during repeated vapor inhalation sessions

BACKGROUND: It is during adolescence that most drinkers initiate ethanol intake, with some of this use being excessive. One possible contributor to the increased ethanol consumption often seen during adolescence in humans and in various animal models is age differences in ethanol sensitivity and tolerance. The present study examined the impact of age on ethanol-related alterations in the autonomic nervous system. METHODS: Sensitivity to the initial ethanol challenge and chronic tolerance as well as acute and protracted withdrawal-like phenomena were assessed in male adolescent and adult Sprague-Dawley rats, using implanted telemetry probes with ethanol delivered via vapor inhalation. RESULTS: Both ages showed similar ethanol-induced tachycardia and activity suppression; however, adolescents were found to be more sensitive than adults to the hypothermic effect of ethanol, data opposite other results from our laboratory and elsewhere using intragastric intubations or intraperitoneal administrations of ethanol. Although little tolerance to ethanol's tachycardic or activity suppressant effects was seen after repeated ethanol inhalation sessions, chronic tolerance to ethanol's hypothermic effect developed faster in adults than in adolescents. A withdrawal-like syndrome, characterized by bradycardia and hypoactivity, typically emerged during the dark phase of the diurnal cycle after ethanol vapor exposure sessions. These effects were observed in animals of both ages, with the bradycardic effect more pronounced in adolescents. CONCLUSIONS: In contrast to results indicating that adolescents may be less sensitive than adults to ethanol's hypothermic effect when ethanol is administered via bolus injection/intubation, adolescents appear more sensitive and develop tolerance to ethanol's hypothermic effects more slowly than adults when ethanol is administered at a more moderate rate via vapor inhalation.     

Effect of stress on the voluntary intake of a sweetened ethanol solution in pair-housed adolescent and adult rats

BACKGROUND: Data regarding the effects of stressors on ethanol intake are mixed. Previous experiments reporting greater voluntary intake of ethanol in adolescent than adult rats have examined intake in isolate-housed animals. Given that the stress of isolate housing may differ ontogenetically as well as confound interpretation of other stressor effects, the present study examined stressor/ethanol interactions among pair-housed adolescent and adult rats. METHODS: Sprague-Dawley male rats were implanted with identification tags that allowed individual monitoring of home cage intake of water and either a 10% (v/v) ethanol solution containing 0.1% (w/v) saccharin or saccharin alone over a 14-day access period. Animals were given zero, one, or eight daily 15-min footshock sessions, with shock-induced freezing and pre-, post-, and recovery corticosterone levels determined on the first and last footshock exposure days. After the access period, withdrawal was assessed with a plus maze, and tolerance to ethanol-induced loss of righting reflex was examined. RESULTS: Nonstressed adolescents drank considerably more sweetened ethanol than did adults, with chronic stress suppressing this adolescent consumption. Ethanol access in adolescents disrupted within-session adaptation to footshock in terms of freezing behavior, although no such disruption was evident at either age when indexed hormonally. Despite relatively high ethanol intakes (up to 6 g/kg/day in the adolescents), no evidence for withdrawal-associated anxiogenesis emerged. Evidence for tolerance was mixed and, to the extent that it was present, was metabolic in nature. CONCLUSIONS: Previous reports of heightened voluntary ethanol intake among adolescent rats are not a function of isolate stress but are evident in pair-housed animals. Adolescents were more sensitive to ethanol/stress interactions than were adults, with the elevated ethanol intake of pair-housed adolescents selectively disrupted by chronic stress, a stress-induced disruption not evident in adults. Likewise, ethanol disrupted behavioral adaptation to the footshock stressor among adolescents but not adults.     

Differential dietary ethanol intake and blood ethanol levels in adolescent and adult rats: effects on anxiety-like behavior and seizure thresholds

Background: Adult rats exhibit increased anxiety‐like behavior after exposure to repeated cycles of chronic ethanol and withdrawal. While adolescent rats have differential responses to both acute and chronic ethanol treatments, the potential differences in the effects of repeated withdrawals in this population have yet to be determined. Methods: Male adult and adolescent rats received three 5‐day cycles of either a 4.5% or 7% ethanol diet (ED) separated by two 2‐day withdrawal periods. Five hours into the final withdrawal, rats were tested for social interaction (SI) deficits (an index of anxiety‐like behavior) and then assessed for seizure thresholds (audiogenic and bicuculline‐induced). Ethanol intake was monitored throughout, and blood ethanol concentrations (BEC) were obtained from a separate group of rats. Results: Adolescent rats have reduced SI during the final withdrawal from either ED and exhibit a greater reduction in SI compared to adult rats when exposed to a 7%ED. Audiogenic seizures were not increased during withdrawal from either ED in adult rats, but adolescent rats that received 7%ED displayed increased seizures. The bicuculline seizure thresholds were decreased in both ages exposed to a 7%ED, but only adolescent rats showed this decreased threshold after 4.5%ED. Ethanol intakes and BECs were higher in adolescent rats compared to similarly treated adults. However, ethanol intakes and BECs were comparable between 4.5%ED‐treated adolescent and 7%ED‐treated adult rats. Conclusions: Behavioral results from the 7%ED‐treated groups suggested that adolescent rats may be more vulnerable to repeated withdrawals from ethanol than adults; however, differences in ethanol intake and BECs may be at least in part responsible. When ethanol intakes and BECs were similar between 4.5%ED‐treated adolescent and 7%ED‐treated adult rats, behavioral effects were not different. Importantly, these data illustrated that adolescent rats can exhibit anxiety and reduced seizure thresholds following this repeated withdrawal paradigm.     

Development of ethanol withdrawal-related sensitization and relapse drinking in mice selected for high- or low-ethanol preference

Background: Previous studies have shown that high alcohol consumption is associated with low withdrawal susceptibility, while at the same time, other studies have shown that exposure to ethanol vapor increases alcohol drinking in rats and mice. In the present studies, we sought to shed light on this seeming contradiction using mice selectively bred for High‐ (HAP) and Low‐ (LAP) Alcohol Preference, first, assessing these lines for differences in signs of ethanol withdrawal and second, for differences in the efficacy of intermittent alcohol vapor exposure on elevating subsequent ethanol intake. Methods: Experiment 1 examined whether these lines of mice differed in ethanol withdrawal‐induced CNS hyperexcitability and the development of sensitization to this effect following intermittent ethanol vapor exposure. Adult HAP and LAP lines (replicates 1 and 2), and the C3H/HeNcr inbred strain (included as a control genotype for comparison purposes) received intermittent exposure to ethanol vapor and were evaluated for ethanol withdrawal‐induced seizures assessed by scoring handling‐induced convulsions (HIC). Experiment 2 examined the influence of chronic intermittent ethanol exposure on voluntary ethanol drinking. Adult male and female HAP‐2 and LAP‐2 mice, along with male C57BL/6J (included as comparative controls) were trained to drink 10% ethanol using a limited access (2 h/d) 2‐bottle choice paradigm. After stable baseline daily intake was established, mice received chronic intermittent ethanol vapor exposure in inhalation chambers. Ethanol intake sessions resumed 72 hours after final ethanol (or air) exposure for 5 consecutive days. Results: Following chronic ethanol treatment, LAP mice exhibited overall greater withdrawal seizure activity compared with HAP mice. In Experiment 2, chronic ethanol exposure/withdrawal resulted in a significant increase in ethanol intake in male C57BL/6J, and modestly elevated intake in HAP‐2 male mice. Ethanol intake for male control mice did not change from baseline levels of intake. In contrast, HAP‐2 female and LAP‐2 mice of both sexes did not show changes in ethanol intake as a consequence of intermittent ethanol exposure. Conclusions: Overall, these results indicate that the magnitude of ethanol withdrawal‐related seizures is inversely related to inherited ethanol intake preference. Additionally, intermittent ethanol vapor exposure appears more likely to affect high‐drinking mice (C57BL/6J and HAP‐2) than low drinkers, although these animals are less affected by ethanol withdrawal.     

Chronic intermittent ethanol exposure during adolescence blocks ethanol-induced inhibition of spontaneously active hippocampal pyramidal neurons

BACKGROUND: Binge alcohol drinking among adolescents has been a serious public health problem. A model of binge alcohol, chronic intermittent ethanol exposure (CIEE), during adolescence significantly attenuates ethanol-induced spatial memory deficits in rats. However, the attenuation was absent following a 12-day ethanol-free period. Since spatial memory is hippocampal dependent, a reduction in ethanol-induced spatial memory impairments may be due to a reduction in the ability of ethanol to inhibit the firing rate of single hippocampal pyramidal neurons following CIEE. METHODS: Beginning on postnatal day 30 (P30), male adolescent Sprague-Dawley rats (Harlan) were administered 5.0 g/kg ethanol (n = 10, CIEE-treated group) or an equivolume saline (n = 10, CISE-treated group) every 48 hours for 20 days. Single hippocampal pyramidal neurons from 5 CIEE-treated rats and 5 CISE-treated rats were recorded on the day following completion of the chronic intermittent exposure procedure (animals now P50). Additionally, neurons from 5 CIEE-treated rats and 5 CISE-treated rats were recorded 12 days after the completion of the chronic intermittent exposure procedure (animals now P62). RESULTS: Ethanol exposure during adolescence completely blocked ethanol-induced inhibition of hippocampal pyramidal neurons in rats that were CIEE exposed. However, the effect of CIEE on hippocampal neurophysiology was time dependent. Specifically, neurons recorded from CIEE-treated rats after a 12-day ethanol-free period had similar maximal inhibition as neurons from CISE-treated animals, although the time to reach inhibition was significantly greater in neurons from CIEE-treated rats. CONCLUSION: Chronic ethanol exposure during adolescence produces a reduction, or tolerance, to ethanol-induced inhibition of hippocampal pyramidal neural activity. Although the tolerance was greatly reversed after a 12-day ethanol-free period, neurons from CIEE animals inhibited slower than neurons from CISE animals. Since the hippocampus is known to be involved not only in spatial memory, but also in many other types of memory formation, the altered hippocampal functions because of CIEE during adolescence should be taken as a serious warning for society.     

Temperature Dependence of Ethanol Depression in Mice: Dose Response

Manipulation of body temperature during intoxication significantly alters brain sensitivity to ethanol. The current study tested the generality of this effect within the hypnotic dose range. Drug naive, male C57BL/6J mice were injected with 3.2, 3.6, or 4.0 g/kg ethanol (20% w/v) and were exposed to 1 of 7 designated temperatures from 13° to 34°C to manipulate body temperature during intoxication. Rectal temperature at return of righting reflex (RORR) was significantly, positively correlated with loss of righting reflex (LORR) duration and significantly, negatively correlated with blood ethanol concentration (BEC) at RORR at all three doses. These results indicate that increasing body temperature during intoxication increased ethanol sensitivity in C57 mice at all three doses tested and demonstrate the generality of temperature dependence across hypnotic doses in these animals. Interestingly, the LORR duration was dose-dependent at each ambient temperature, but the degree of body temperature change and the BEC at RORR were not dose-dependent. Overall, these results emphasize the importance of body temperature as a variable in ethanol research.     

Increased ethanol drinking after repeated chronic ethanol exposure and withdrawal experience in C57BL/6 mice

BACKGROUND: The development of dependence may have significant motivational consequences regarding continued use and abuse of ethanol. We have developed a mouse model of ethanol dependence and repeated withdrawals that demonstrates sensitization of seizures and other symptoms of withdrawal. It is unclear whether such experience influences ethanol drinking behavior. The present series of experiments were designed to examine whether repeated cycles of chronic ethanol exposure and withdrawal has an impact on subsequent motivation to voluntarily self-administer ethanol. METHODS: With the use of a modified sucrose-fading procedure, adult male C57BL/6J mice were trained to drink 15% (v/v) ethanol in a limited access procedure (2 hr/day). The animals were not food or water deprived at any time during the experiments. Once stable baseline intake was established, mice were exposed to four cycles of 16 hr of ethanol vapor (or air) in inhalation chambers separated by 8-hr periods of withdrawal. At 32 hr after the last cycle of ethanol exposure, all mice were tested for ethanol intake under limited access conditions for 5 consecutive days. The animals then received a second series of chronic ethanol exposure and withdrawal followed by another 5-day test period for ethanol drinking. RESULTS: Stable daily baseline intake was established in mice that drank 15% ethanol combined with 5% sucrose (experiment 1), 15% ethanol alone (experiment 2), 5% sucrose alone (experiment 3), or 15% ethanol when presented as a choice with water (experiment 4). After repeated cycles of chronic ethanol exposure and withdrawal experience, consumption of ethanol solutions increased over baseline levels and in comparison with control (air-exposed) groups. However, sucrose consumption did not change in mice that were trained to drink 5% sucrose. The increase in ethanol consumption after chronic ethanol exposure and withdrawal experience resulted in a significant increase in resultant blood ethanol levels. CONCLUSIONS: Once the positive reinforcing properties of ethanol were established, chronic ethanol exposure and withdrawal experience resulted in a significant increase in voluntary ethanol drinking that yielded a >2-fold increase in resultant blood ethanol levels. This increase in ethanol intake occurred whether ethanol was presented in combination with sucrose, alone (unadulterated), or as a choice with tap water. Furthermore, this effect seems to be selective for ethanol in that animals that were trained to drink a sucrose solution did not exhibit a change in their intake after similar chronic ethanol exposure. As such, this model may be useful in studying the mechanisms and conditions in which chronic ethanol treatment influences motivation to resume drinking after a period of abstinence (relapse).     

Role of novelty and ethanol history in locomotor stimulation induced by binge-like ethanol intake

Background: The acute locomotor effects of voluntary ethanol (EtOH) intake in mice (stimulation/sedation) might be important behavioral indicators of an animals’ propensity to engage in EtOH consumption and/or EtOH seeking behaviors. Using a binge‐like EtOH intake model dubbed “Drinking‐in‐the‐Dark (DID),” we recently observed home cage locomotor stimulation in C57BL/6J mice during an acute EtOH intake session, but acute home cage locomotor sedation following repeated EtOH exposures. To determine the role of novelty and/or EtOH history on these previously described locomotor effects, and to determine the relationship between these variables on locomotor activity immediately following DID intake, we conducted 2 separate experiments. Methods: In experiment 1, mice were given access to either EtOH or water, and locomotor activity was monitored immediately afterwards. In experiment 2, mice were given 13 days access to EtOH or water solution while home cage locomotor activity was monitored. On the 14th day, half of the water consuming animals received EtOH access for the first time. On the 15th day, all animals received EtOH access, and locomotion was assessed afterwards in locomotor activity testing chambers. Results: In experiment 1, locomotor activity following DID was positively associated with EtOH intake and blood EtOH concentrations (BECs). In experiment 2, the group that received EtOH for the first time on the 14th day did not display locomotor stimulation. Locomotor activity following DID EtOH intake was positively associated with BECs in all groups regardless of EtOH history. Conclusions: These results suggest that (i) DID‐induced locomotor stimulation in the home cage may involve relative familiarity with the DID procedures, and (ii) locomotor stimulation immediately following DID is directly related to the relative concentration of EtOH in blood; an effect that is not altered by prior EtOH history. These data add new evidence of the pharmacological actions of binge‐like EtOH intake, and provide a basis by which we may explore the motivation and consequences of such binge consumption.     

Acute, rapid, and chronic tolerance during ontogeny: observations when equating ethanol perturbation across age

Background: Sensitivity to the motor‐impairing and hypnotic effects of ethanol (EtOH) increases notably during development. Less is known, however, about the ontogeny of EtOH tolerance and the ontogenetic relationship among different types of tolerance. Consequently, we compared the ontogenetic development of acute, rapid, and chronic tolerance to EtOH‐induced motor impairment and hypothermia in a swim task. Methods: Preweanling, adolescent, and adult female and male Sprague‐Dawley rats were given chronic saline (control group), five daily EtOH exposures before EtOH on test day (chronic group), one EtOH exposure before test day (rapid group), or EtOH exposure only on test day (acute groups). Separate groups of animals in the acute groups were tested at 15, 60, or 105 min after injection to estimate acute tolerance development via calculating slopes of the linear regression of impairment relative to brain alcohol levels at each postinjection interval. Initial EtOH perturbation of swim performance was equated across age by varying EtOH dose. Results: Acute tolerance was evident to the motor‐impairing effects of EtOH at all ages. When impairment was indexed relative to brain alcohol levels, rapid and chronic tolerance to the motor‐impairing effects of EtOH on latency to reach the start was seen across age, although this tolerance tended to be more pronounced in adults. Somewhat different ontogenetic patterns of tolerance development were observed with EtOH‐induced hypothermia, a dependent measure for which EtOH perturbation was not equated across age. Conclusions: The degree of initial perturbation by EtOH seems to be an important predictor of tolerance expression during ontogeny. That is, ontogenetic profiles of tolerance development differ significantly when EtOH‐induced motor impairment is equated across age rather than dose of EtOH administered . The role of target response measures and context stress should also be considered when exploring ontogenetic expression of EtOH tolerance.     

Quantitative trait loci mapping for ethanol sensitivity and neurotensin receptor density in an F2 intercross derived from inbred high and low alcohol sensitivity selectively bred rat lines

BACKGROUND: Genetic variance in initial sensitivity to ethanol has been implicated as a risk factor for the development of alcoholism. Identification of the genes that confer differential initial sensitivity is an important goal for the development of new treatment strategies and for a comprehensive understanding of the mechanism of ethanol's action. Quantitative trait loci (QTL) mapping for initial sensitivity and other ethanol-related behavioral traits in model organisms has become an important first step for the ultimate identification of genes that contribute to variation in ethanol responses. METHODS: An F(2) intercross was made from the Inbred High and Low Alcohol Sensitivity rat lines (IHAS and ILAS). The F(2) rats were tested for duration of the loss of righting reflex test (LORR); blood ethanol concentration at regain of righting reflex (BECrrr); BEC at the first time to reach criterion on the rotarod after 1.6 g/kg of ethanol (BEC1); acute functional tolerance on the rotarod (AFT); and high-affinity neurotensin receptor (NTR1) density in the nucleus accumbens (NAc), caudate putamen (CP), and ventral midbrain (VMB). A full genome scan with an average marker spacing of 16.8 cM for interval QTL mapping was conducted on the F(2) rats (N = 363). RESULTS: Seven significant or suggestive QTL were detected for LORR, one for BECrrr, three for BEC1, two for NTR1 binding in the CP, and one for binding in the NAc, but none were mapped for AFT or NTR1 binding density in the VMB. Effect size of the seven LORR QTL, the trait for which the parental strains were selected, ranged from 3 to 4%, with all accounting for approximately 22% of the total phenotypic variation. One of the LORR QTL on chromosome 2 (approximately 87 cM) was significant, and a second QTL on chromosome 5 (approximately 37 cM) was suggestive for both LORR and BECrrr. CONCLUSIONS: The results indicate that segregating populations derived from the IHAS and ILAS strains can be used for mapping ethanol sensitivity QTL. The chromosome 2 LORR QTL may confer variation in ethanol metabolism, whereas the chromosome 5 LORR/BECrrr QTL likely mediates central nervous system ethanol sensitivity. The small number or absence of QTL for BEC1, AFT, and NTR1 receptor density suggests that genetic variation for these traits is minimal in the IHAS/ILAS strains and/or the effect size of QTL for these traits is too small to be mapped efficiently in this sample of F(2) rats. The ultimate identification of genes underlying these alcohol sensitivity QTL will contribute to our understanding of the actions of alcohol in the central nervous system if not to a deeper understanding of the genetic risk factors for alcoholism.     

Ethanol-Induced Social Facilitation in Adolescent Rats: Role of Endogenous Activity at Mu Opioid Receptors

Background: Ethanol consumption is considerably elevated during adolescence. Attractiveness of alcohol for humans during the adolescent developmental period is based, in part, on its ability to induce social facilitation—a facilitation of social interactions not only evident in human adolescents but also in adolescent rats. Endogenous opioid systems are among the multiple neural systems implicated in the behavioral and reinforcing effects of ethanol and may play a substantial role in modulating stimulatory effects of low doses of ethanol on social behavior during adolescence. This possibility was explored in the present study through the use of an animal model of peer‐directed social behavior. Methods: Sprague–Dawley rats were challenged early in adolescence with saline or ethanol intraperitoneally (i.p.), placed into an individual holding cage for 30 minutes, and then tested in a familiar situation with a nonmanipulated partner of the same age and sex. In Experiment 1 , each test subject was injected subcutaneously with one of the three doses of a nonselective opioid antagonist naloxone (0, 0.05, and 0.1 mg/kg), 5 minutes prior to the social interaction test and 25 minutes following challenge with saline or ethanol (0.5 g/kg), whereas in Experiment 2 animals were challenged with one of the six doses of ethanol (0, 0.25, 0.5, 0.75, 1.0, and 1.25 g/kg) prior to injection of either saline or naloxone (0.05 mg/kg). In Experiment 3 , animals were pretreated i.p. with the selective μ‐opioid antagonist CTOP (0, 0.01, 0.025, 0.05, and 0.1 mg/kg) 30 minutes prior to challenge with saline or ethanol (0.5 g/kg). Results: Low doses of ethanol (0.5 and 0.75 g/kg) produced social facilitation, as indexed by significant increases in play fighting and social investigation. Both doses of naloxone and the three highest doses of CTOP blocked the stimulatory effects of ethanol on play fighting but not on social investigation. These effects were not associated with alterations in ethanol pharmacokinetic properties or with shifts in the biphasic ethanol dose–response curve. Conclusions: Ethanol‐induced facilitation of social play behavior seen in adolescent animals is mediated in part through ethanol‐induced release of endogenous ligands for the μ‐opioid receptor or an ethanol‐associated enhancement of sensitivity of these receptors for their endogenous ligands.     

Effects of ethanol and ethanol withdrawal on nociception in rats

The effect of acute and chronic administration of ethanol and ethanol withdrawal on a radiant heat tail-flick assay of nociception was examined in rats. Acute administration of ethanol (2.0 g/kg, i.p.) produced peak antinociception (68% of maximum) by 30 min, and effects were gone by 120 min. Cumulative doses of ethanol (0.5-2.0 g/kg, i.p.) produced dose-dependent increases in latencies to 49% of maximum. During chronic administration, a liquid diet containing ethanol (6.5%) was given for 10 days. Tail-flick latencies were measured on day 0 (baseline), day 2, 4, 6, 8, and 10 of chronic ethanol and at 3, 6, 12, and 36 hr after removal of ethanol. To test for behavioral tolerance, both between- and within-group designs were used. In both between- and within-group experiments, the antinociceptive effects of chronic ethanol peaked by day 4 of exposure to the liquid diet, and tolerance developed by day 10. When the liquid diet was removed, hyperalgesia was detected at 6 and 12 hr after withdrawal, and was gone by 36 hr after withdrawal. When cumulative doses of ethanol (0.5-2.0 g/kg) were administered starting 12 hr after withdrawal, ethanol (0.5 g/kg) fully reversed the hyperalgesia induced by ethanol withdrawal, even though this dose was without antinociceptive effect in the absence of withdrawal. Higher doses of ethanol during ethanol withdrawal did not increase tail-flick latencies over baseline. In summary: (1) ethanol produces antinociception when administered acutely or chronically; (2) tolerance to the antinociceptive effects develops during chronic administration; (3) ethanol withdrawal induced hyperalgesia, which was reversed by ethanol; and (4) repeated testing did not produce behavioral tolerance.     

Adolescent and adult heart rate responses to self-administered ethanol

Background: Despite the fact that adolescent rats have repeatedly been found to consume more ethanol than adult rats in a variety of ethanol access paradigms, the exact cause of the increase in ethanol consumption during adolescence is not known. One possibility is that age differences in sensitivity to ethanol’s rewarding effects may contribute to the elevated intake seen among adolescents. Human studies have shown that autonomic effects of ethanol, particularly ethanol‐induced tachycardia, are correlated with the positive hedonic properties of the drug and, hence, may serve as a biomarker for reward. Methods: In this experiment, a limited‐access self‐administration paradigm was used to examine the autonomic effects of ethanol in outbred male adolescent and in adult Sprague‐Dawley rats under circumstances likely to reveal the rewarding value of ethanol. Results: The results indicated that voluntary ethanol consumption was greater in adolescent than adult rats and that only adolescents consumed enough of the saccharin‐sweetened ethanol solution to show a tachycardic effect greater than that seen in response to saccharin alone. Conclusions: To the extent that these tachycardic properties of ethanol are associated with the rewarding/hedonic properties of ethanol as previously reported in humans, these findings support the suggestion that adolescent animals may have found the ethanol‐containing solution to be more rewarding than the saccharin solution. A similar effect was not seen in adults, findings consistent with the notion that adult rats may not consume enough ethanol under these circumstances to experience its positive rewarding properties.     

The effects of gonadectomy on age- and sex-typical patterns of ethanol consumption in Sprague-Dawley rats

Background: Ethanol intake levels characteristic of adult males and females emerge postpubertally. The present set of experiments examined the consequences of prepubertal and adult gonadectomies to explore whether the presence of gonadal hormones at puberty exerts organizational influences and/or plays an activational role in age‐ and sex‐typical patterns of ethanol consumption. Methods: Male and female Sprague–Dawley rats were gonadectomized (GX), received sham gonadectomy (SH), or were left nonmanipulated (NM) at 1 of 2 ages, either prepubertally on postnatal day (P) 23 (early) or postpubertally in adulthood on P70 (late). Early surgery animals were tested for ethanol consumption either during adolescence (P28 to 39) or in adulthood at the same age that late surgery animals were tested (P75 to 86). Voluntary ethanol consumption was indexed using a 2‐hour limited‐access paradigm, with access to 2 bottles: one containing water and the other a sweetened ethanol solution. Results: Age of GX did not impact patterns of ethanol consumption. Removal of testicular hormones in males, regardless of age of removal, elevated consumption levels in adulthood to female‐typical levels. Ovariectomy did not have notable effects on ethanol drinking in females. Ethanol intake and preference of early SH males were significantly greater than those of both late SH and NM males. Removal of the gonads prior to puberty did not influence ethanol drinking or preference during adolescence in either males or females. Conclusions: These results suggest that testicular hormones play an activational role in lowering ethanol intake and preference of adult male rats. Pubertal hormones, in contrast, were found to exert little influence on ethanol drinking or preference during adolescence, although the effect of surgical manipulation itself during development was found to exert a long‐lasting facilitatory effect on ethanol consumption in adulthood.