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

Biol Psychiatry. 2019 Jun 1;85(11):904-914. doi: 10.1016/j.biopsych.2018.12.021. Epub 2019 Jan 14.


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: Adolescent alcohol; Amygdala; Anxiety; Arc; Enhancer RNA; epigenetic.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Age Factors
  • Amygdala / drug effects
  • Amygdala / metabolism
  • Animals
  • Anxiety / genetics*
  • Cytoskeletal Proteins / biosynthesis
  • Cytoskeletal Proteins / genetics*
  • Enhancer Elements, Genetic / genetics*
  • Epigenesis, Genetic / drug effects
  • Jumonji Domain-Containing Histone Demethylases / antagonists & inhibitors
  • Male
  • Microinjections
  • Nerve Tissue Proteins / biosynthesis
  • Nerve Tissue Proteins / genetics*
  • Oligonucleotides, Antisense / administration & dosage
  • Oligonucleotides, Antisense / pharmacology
  • RNA, Small Interfering / administration & dosage
  • RNA, Small Interfering / pharmacology
  • RNA, Small Untranslated / biosynthesis*
  • RNA, Small Untranslated / genetics
  • Rats


  • Cytoskeletal Proteins
  • Nerve Tissue Proteins
  • Oligonucleotides, Antisense
  • RNA, Small Interfering
  • RNA, Small Untranslated
  • activity regulated cytoskeletal-associated protein
  • Jumonji Domain-Containing Histone Demethylases
  • Kdm6b protein, rat