Circadian Phase Determines Effects of Repeated Ethanol Vapor Exposure and Withdrawal on Body Temperature and Activity Rhythms of Male Mice

Chronic alcohol consumption contributes to fatty liver disease. Our studies revealed that the hepatic circadian clock is disturbed in alcohol-induced hepatic steatosis, and effects of chronic alcohol administration upon the clock itself may contribute to steatosis. We extended these findings to explore the effects of chronic alcohol treatment on daily feeding and locomotor activity patterns. Mice were chronically pair-fed ad libitum for 4 weeks using the Lieber-DeCarli liquid diet, with calorie-controlled liquid and standard chow diets as control groups. Locomotor activity, feeding activity, and real-time bioluminescence recording of PERIOD2::LUCIFERASE expression in tissue explants were measured. Mice on liquid control and chow diets exhibited normal profiles of locomotor activity, with a ratio of 22:78% day/night activity and a peak during early night. This pattern was dramatically altered in alcohol-fed mice, marked by a 49:51% ratio and the absence of a distinct peak. While chow-diet fed mice had a normal 24:76% ratio of feeding activity, with a peak in the early night, this pattern was dramatically altered in both liquid-diet groups: mice had a 43:57% ratio, and an absence of a distinct peak. Temporal differences were also observed between the two liquid-diet groups during late day. Cosinor analysis revealed a ~4-h and ~6-h shift in the alcohol-fed group feeding and locomotor activity rhythms, respectively. Analysis of hepatic PER2 expression revealed that the molecular clock in alcohol-fed and control liquid-diet mice was shifted by ~11 h and ~6 h, respectively. No differences were observed in suprachiasmatic nucleus explants, suggesting that changes in circadian phase in the liver were generated independently from the central clock. These results suggest that chronic alcohol consumption and a liquid diet can differentially modulate the daily rhythmicity of locomotor and feeding behaviors, aspects that might contribute to disturbances in the circadian timing system and development of hepatic steatosis.

Section: Discussionmentioning

confidence: 99%

Dissociation between diurnal cycles in locomotor activity, feeding behavior and hepatic PERIOD2 expression in chronic alcohol-fed mice

Zhou

Werner

Lee

et al. 2015

“…Hyperthermia is often observed in humans during ethanol withdrawal and it is a risk factor for later development of delirium tremens (Hall & Zador, 1997; Monte et al, 2009). Studies in animal models have shown that either hyperthermia or hypothermia can be shown, depending on the circadian phase (Damaggio et al, 2014a,b). We have also shown disruption of thermoregulation during withdrawal from vapor inhalation (Crawshaw et al, 1994).…”

Section: Methodsmentioning

confidence: 99%

“…In WD Battery 1, we included body temperature as the last behavior measured (WD Hr 26). We estimated basal body temperature in this experiment one week after the experiment ended, at the same time of day as the temperature assessment during withdrawal, in order to avoid potential circadian confounds (Tankersley et al, 2002; Damaggio et al, 2014a). In WD Batteries 2 and 3, we assessed basal body temperatures 1 to 4 days before placing mice into the vapor inhalation chambers.…”

Section: Methodsmentioning

confidence: 99%

An alcohol withdrawal test battery measuring multiple behavioral symptoms in mice

Metten

Schlumbohm

Huang

et al. 2018

Despite acceptance that risk for alcohol-use disorder (AUD) has a large genetic component, the identification of genes underlying various components of risk for AUD has been hampered in humans, in part by the heterogeneity of expression of the phenotype. One aspect of AUD is physical dependence. Alcohol withdrawal is a serious consequence of alcohol dependence with multiple symptoms, many of which are seen in multiple species, and can be experienced over a wide-ranging time course. In the present three studies, we developed a battery of withdrawal tests in mice, examining behavioral symptoms from multiple domains that could be measured over time. To permit eventual use of the battery in different strains of mice, we used male and female mice of a genetically heterogeneous stock developed from intercrossing eight inbred strains. Withdrawal symptoms were assessed using commonly used tests after administration of ethanol in vapor for 72 continuous hours. We found significant effects of ethanol withdrawal versus air-breathing controls on nearly all symptoms, spanning 4 days following ethanol vapor inhalation. Withdrawal produced hypothermia, greater neurohyperexcitability (seizures and tremor), anxiety-like behaviors using an apparatus (such as reduced transitions between light and dark compartments), anhedonia (reduced sucrose preference), Straub tail, backward walking, and reductions in activity; however, there were no changes in thermal pain sensitivity, hyper-reactivity to handling, or anxiety-like emergence behaviors in other apparatus. Using these data, we constructed a refined battery of withdrawal tests. Individual differences in severity of withdrawal among different tests were weakly correlated at best. This battery should be useful for identifying genetic influences on particular withdrawal behaviors, which should reflect the influences of different constellations of genes.

“…Many experimental paradigms have been used for chronic ethanol administration, including daily ethanol injections or intra-gastric lavage, daily ethanol vapor exposure, continuous ad libitum ethanol (5-25%) drinking (either in free-choice or a forced-consumption situation), or fixed interval drinking, with ethanol available at specified times each 24 h (e.g., chronic intermittent drinking, or drinking-in-the-dark). The choice of paradigm used often depends on whether the focus of the study is on regulating the exact amount and/or timing of ethanol administered vs. allowing subjects to control their own ethanol consumption (Kliethermes et al, 2005;Logan et al, 2010;Rosenwasser et al, 2010;Iancu et al, 2013;Pisu et al, 2011;Ahmed, 2012;Damaggio and Gorman, 2013). …”

Section: Evidence Of Ethanol Tolerance In the Scn Circadian Systemmentioning

confidence: 99%

Assessing ethanol's actions in the suprachiasmatic circadian clock using in vivo and in vitro approaches

Prosser

Glass

2015

Research over the past decade has demonstrated substantial interactions between the circadian system and the processes through which alcohol affects behavior and physiology. Here we summarize the results of our collaborative efforts focused on this intersection. Using a combination of in vivo and in vitro approaches, we have shown that ethanol affects many aspects of the mammalian circadian system, both acutely as well as after chronic administration. Conversely, we have shown circadian influences on ethanol consumption. Importantly, we are beginning to delve into the cellular mechanisms associated with these effects. We are also starting to form a picture of the neuroanatomical bases for many of these actions. Finally, we put our current findings into perspective by suggesting new avenues of inquiry for our future efforts.