Behavioral characterization of withdrawal following chronic voluntary ethanol consumption via intermittent two‐bottle choice points to different susceptibility categories

Abstract

Background Alcohol is among the most commonly abused drugs worldwide. Cessation of chronic alcohol consumption can result in the appearance of withdrawal symptoms that commonly promote relapse in individuals with alcohol use disorder (AUD). Thus, preclinical models of voluntary alcohol consumption, in which animals manifest spontaneous signs of withdrawal after alcohol cessation, can be useful for studying AUD and its treatment. The intermittent two‐bottle choice paradigm (I2BC) has been used extensively to examine alcohol intake in rodents. However, previous studies have reported conflicting observations regarding its potential to result in the spontaneous manifestation of withdrawal upon alcohol cessation. Methods We employed a battery of behavioral tests to examine the emergence of affective and physical signs of withdrawal in female and male mice exposed to alcohol in the I2BC for 10 weeks. Specifically, mice of both sexes undergoing 24‐h withdrawal from the I2BC were tested for physical signs of withdrawal, anxiety‐like behavior in the open field arena (OFA) and elevated plus maze (EPM), and anxiety/compulsive‐like behavior in the marble burying test (MBT). The main outcomes from these tests were combined into a behavioral severity score to describe the overall behavioral phenotype. Results Both female and male mice undergoing withdrawal from the I2BC displayed elevated physical signs of withdrawal and anxiety‐associated behavior in the EPM and MBT. Analysis of the overall behavioral severity score revealed more severe phenotypes in female and male mice undergoing withdrawal from the I2BC than controls. Additionally, stratification of the mice based on severity scores demonstrated a differential distribution of severities between the exposure groups. Conclusions We confirmed that a significant fraction of mice chronically exposed to alcohol in the I2BC display spontaneous withdrawal. In addition, we showed that computing a severity score from a combination of behavioral metrics can be useful in preclinical research to model evaluation tools used in patients with AUD.

In utero exposure to morphine leads to sex-specific behavioral alterations that persist into adulthood in cross-fostered mice

“…Given the well-documented interactions between alcohol and the opioid system (Arias & Kranzler, 2008; Berrettini, 2013; Gianoulakis, 2001; Gianoulakis et al, 1989; Job et al, 2007; Oslin, Berrettini, & O’Brien, 2006), we next wanted to assess alcohol use risk in offspring maternally exposed to morphine. The offspring tested in the battery of behavioral tests during adolescence were allowed to mature into adulthood and were then evaluated in an ethanol intermittent two-bottle choice (I2BC) paradigm (Figure 6a), which has been used to assess voluntary ethanol intake (Carnicella et al, 2014; Hwa et al, 2011; Quijano Cardé & De Biasi, 2022; Quijano Cardé et al, 2021). A three-way ANOVA revealed a main effect of sex where female mice (regardless of treatment) drank significantly more ethanol than male mice, so data and analyses are presented separately for each sex.…”

“…if locomotion is changed then that might influence time spent in a particular zone and affect anxiety-like measures). Similar to O'Neal et al (2020) andQuijano Cardé et al (2022), z-scores were calculated for each behavioral measure. Briefly, the group mean (µ 1 ) for each behavioral measure was subtracted from the raw individual value (x 1 ) for each offspring for that behavior, and then divided by the group standard deviation, (…”

“…Mice that were previously analyzed for adolescent baseline behaviors were examined for ethanol drinking behavior during adulthood using the ethanol I2BC, as previously described (Carnicella, Ron, & Barak, 2014;Hwa et al, 2011;Quijano Cardé & De Biasi, 2022;Quijano Cardé, Perez, Feinn, Kranzler, & De Biasi, 2021). Offspring (at least 2 months of age) were habituated to being single-housed and were exposed to two 50 mL bottles of filtered water for at least one week in the home cage.…”

Sex differences in binge alcohol drinking and the behavioral consequences of protracted abstinence in C57BL/6J mice

“…Indeed, the results of previous work on the affective behavioral consequences of chronic alcohol abstinence in C57BL/6J mouse models are variable as to time into abstinence tested and paradigms utilized, as well as highly male-dominated (16). Many studies have shown that males show increased avoidance, anhedonia, and compulsive behaviors in early abstinence (>1 week) following chronic alcohol across vapor inhalation (34)(35)(36)(37)(38)(39), continuous access (21,40), intermittent drinking (13), and DID (18,22,41,42). Behavior during protracted abstinence (<1 week) in males is somewhat variable, with increased avoidance following chronic vapor (17) and DID (15,18), with no effects following continuous (20,21) and on most measures during intermittent access (10), and fairly consistent increased anhedonia across alcohol vapor (17,19) and continuous drinking (20,21,40), with some exceptions for DID (18,23).…”

“…Nonetheless, the complexities of binge drinking behavior remain poorly understood in females, including in rodent models such as the commonly used C57Bl/6J mouse strain that has been employed robustly in males. In particular, it is unclear whether females consume more than males due to differences in alcohol preference or motivated voluntary consumption, as the studies performed to date report a mix of results showing elevated female preference for at least some time points of alcohol exposure (10,11) or no sex differences (9,(13)(14)(15), depending on the alcohol exposure paradigm and length.…”