Chronic Alcohol Consumption Alters Home-Cage Behaviors and Responses to Ethologically Relevant Predator Tasks in Mice

Abstract

Alcohol use disorders (AUD) are the most prevalent substance use disorders worldwide. Considering recent reports indicating an increase in alcohol use particularly in females, it is vital to understand how alcohol history impacts behavior. Animal model research on withdrawal-associated affective states tends to focus on males, forced alcohol paradigms, and a few traditional anxiety/stress tests. While this has been essential, heavy alcohol use triggers adverse withdrawal-related affective states that can influence how people respond to a large variety of life events and stressors. To this end, we show that behaviors in the home-cage, open field, looming disc, and robogator predator threat task, which vary in task demand and intensity, are altered in mice with a history of voluntary alcohol consumption. In alcohol-exposed males, behaviors in the home cage, a low anxiety baseline environment, suggest increased vigilance/exploration. However, in the open field and robogator task, which induce heightened arousal and task demands, a more hesitant/avoidant phenotype is seen. Female alcohol mice show no behavioral alterations in the home cage and open field test, however, in the looming disc task, which mimics an overhead advancing predator and forces a behavioral choice, we see greater escape responses compared to water controls, indicative of active stress coping behaviors. This suggests females may begin to show alcohol-induced alterations as task demands increase. To date, few drugs have advanced past clinical trials for the treatment of AUD, and those that have are predominately used in life-threatening situations only. No treatments exist for ameliorating negative withdrawal related states, which could aid in harm reduction related to heavy alcohol use. Understanding how withdrawal alters a variety of behavioral responses that are linked to stress coping can widen our understanding of alcohol abuse and lead us closer to better therapeutics to help individuals with AUD.

Impact and role of hypothalamic corticotropin releasing hormone neurons in withdrawal from chronic alcohol consumption in female and male mice

“…Post hoc Šidák corrected multiple comparisons test did not reveal any significant differences (30pA: p = 0.065 and 40pA: p = .108). We only found differences between alcohol and water exposed males in the input resistance ( We had previously shown (Neira et al, 2022) that female mice in 6-8hr acute withdrawal had increased escape behaviors in the looming disc test, therefore, we hypothesized that inhibiting PVN CRH neurons will restore female behavior in this test. First, we established that increased escape responses compared to mCherry water controls in 6day alcohol withdrawal mCherry mice were also seen in the looming disc test (Fig 5A In fact, Chi-square analysis between mCherry water and alcohol mice during Escape, Freeze, or neither escape/freeze behavioral responses to loom revealed significant differences in behavior (Fig 5B X 2 (1, N =130) = 11.3502, p =.003), with alcohol mCherry mice displaying greater escape responses.…”

“…After habituation, the home-cage with only the bedding and mouse was placed in a ~60 lux low-light sound attenuated chamber that contained an overhead camera for filming behavior. Rearing, digging, grooming, and locomotion were analyzed for a 15minute period using DeepLabCut and SimBA mediated automatic analysis as previously discussed (Neira et al, 2022).…”

“…Female rodents show a greater activation of the HPA-axis stress system to different stressors, including acute alcohol challenges, and display more active stress responses in behavioral tasks that activate this system (Bangasser et al, 2018;Kudielka & Kirschbaum, 2005;Palanza, 2001;Palanza & Parmigiani, 2017;Rivier, 1999), perhaps suggesting interactions between chronic alcohol and stress response. In fact, females consume high levels of alcohol in a two-bottle choice (2-BC) voluntary alcohol paradigm, and compared to water controls, display greater active escape behaviors during acute withdrawal in the looming disc test that mimics an approaching predator (Neira et al, 2022). PVN CRH neurons are activated prior to escape initiation in this task, and optogenetic inhibition of these neurons reduces escape behavior, indicating that PVN CRH neurons are necessary for active escape responses to the looming disc (Daviu et al, 2020).…”

Using deep learning to study emotional behavior in rodent models

“…The advent of machine learning and deep learning enables the simultaneous specialization and standardization of behavior classification processes across neuroscience research. Previous studies have utilized various deep learning techniques to look at behaviors that correlate with pain states (Bohic et al, 2021 ), social behavior in stressed mice (Rodriguez et al, 2020 ; Lee et al, 2022 ), depressive behavior in stressed mice (Rivet-Noor et al, 2022 ), and reward behavior in mice models of food seeking (Zhang et al, 2022 ) or drug and alcohol abuse (Campos-Ordoñez et al, 2022 ; Neira et al, 2022 ). This allows behavioral neuroscientists to explore behavioral nuances in a more generalizable way (Goodwin et al, 2022 ).…”

Chronic Alcohol Drinking Drives Sex-Specific Differences in Affective Behavior and Medial Prefrontal Cortex Activity in CRF1:Cre:Tdtomato Transgenic Rats

“…Others have found that various aspects of EtOH withdrawal (including increased anxiety-like behavior, seizures, and EtOH intake during withdrawal) increase with repeated EtOH withdrawal cycles (Becker, 1996; Holter et al, 1998; Lopez and Becker, 2005; Overstreet et al, 2004; Rimondini et al, 2003; Zhang et al, 2007). Another possibility is that the environment was too anxiogenic, creating a ceiling effect of anxiety-as suggested by a recent study of testing outside the homecage(Neira et al, 2022). Future studies will investigate negative affect as it occurs in the home cage, as it may be more indicative of withdrawal behaviors.…”