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Adolescent Alcohol Exposure Epigenetically Suppresses Amygdala Arc Enhancer RNA Expression to Confer Adult Anxiety Susceptibility

  • Evan J. Kyzar
    Affiliations
    Center for Alcohol Research in Epigenetics, Department of Psychiatry, University of Illinois at Chicago, Chicago, Illinois

    Jesse Brown Veterans Affairs Medical Center, Chicago, Illinois
    Search for articles by this author
  • Huaibo Zhang
    Affiliations
    Center for Alcohol Research in Epigenetics, Department of Psychiatry, University of Illinois at Chicago, Chicago, Illinois

    Jesse Brown Veterans Affairs Medical Center, Chicago, Illinois
    Search for articles by this author
  • Subhash C. Pandey
    Correspondence
    Address correspondence to Subhash C. Pandey, Ph.D., Center for Alcohol Research in Epigenetics, Department of Psychiatry, University of Illinois at Chicago, 1601 West Taylor Street (m/c 912), Chicago, IL 60612.
    Affiliations
    Center for Alcohol Research in Epigenetics, Department of Psychiatry, University of Illinois at Chicago, Chicago, Illinois

    Department of Anatomy and Cell Biology, University of Illinois at Chicago, Chicago, Illinois

    Jesse Brown Veterans Affairs Medical Center, Chicago, Illinois
    Search for articles by this author

      Abstract

      Background

      Adolescent intermittent ethanol (AIE) exposure is an emerging risk factor for adult psychopathology, such as anxiety disorders. Enhancer RNAs (eRNAs) are short noncoding RNAs transcribed from enhancer regions that regulate synaptic plasticity–associated gene expression, including Arc, but their role in AIE-induced susceptibility to anxiety in adulthood is unknown.

      Methods

      Rats were exposed to AIE (ethanol exposure 2 days on/off) or intermittent normal saline during postnatal days 28 to 41 and allowed to grow to adulthood for analysis of behavior and biochemical measures. Some AIE rats and rats with intermittent normal saline exposure were exposed to an acute challenge with ethanol in adulthood. Cohorts of alcohol-naïve adult rats were cannulated in the central nucleus of amygdala and infused with either Kdm6b small interfering RNA or an antisense locked nucleic acid oligonucleotide specific to Arc eRNA before behavioral and biochemical analysis.

      Results

      AIE adult rats displayed heightened anxiety and decreased Arc eRNA expression, which is regulated epigenetically through decreased Kdm6b expression. This triggered condensed chromatin at the synaptic activity response element site and promoter of the Arc gene, facilitating increased negative elongation factor binding to the Arc promoter and decreasing Arc expression in the amygdala. Knockdown of Kdm6b or Arc eRNA expression in the central nucleus of amygdala provoked anxiety in alcohol-naïve adult rats and recapitulated the molecular and epigenetic phenotypes of AIE.

      Conclusions

      These data suggest that eRNA regulation via epigenetic reprogramming in the amygdala, particularly at the Arc synaptic activity response element site, contributes to adult anxiety after adolescent alcohol exposure.

      Keywords

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      Linked Article

      • Connecting the Dots: Adolescent Alcohol, Enhancer RNA, and Anxiety
        Biological PsychiatryVol. 85Issue 11
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          The overwhelming drug of choice for abuse by adolescents is alcohol, and when they drink, it is often in intermittent binges, consuming more than four drinks in a few hours (1). Considerable epidemiological evidence documents that adolescent exposure to ethanol can increase the risk for drug dependence, affective disorders, or cognitive impairment in adulthood (2). Similar results from adolescent ethanol exposure have been found in animal models (3) and also occur with other drugs of abuse (4). Such findings document that adolescence is a critically sensitive developmental period whereby ethanol can evoke long-lasting changes in brain function, possibly due to alterations in ongoing synaptic pruning and maturation of myelinated fibers.
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