[R01] Epigenetic regulation of stress-potentiated ethanol drinking
Ente: National Institute on Alcohol Abuse and Alcoholism
Scadenza: 2030-04-30
Importo max: 398.823 EUR
Paese: US
Descrizione
Epigenetic Regulation of Stress-Potentiated Ethanol Drinking
PROJECT SUMMARY/ABSTRACT
A common mechanism that regulates both stress-sensitivity and alcohol use is epigenetic regulation of gene
transcription. Of particular importance in substance-use disorders is G9a, a histone methyltransferase implicated
in models of alcohol use disorder (AUD). G9a in the nucleus accumbens (NAc) regulates drinking, but its
mechanisms are unknown. The long-term goal of this project is to elucidate the mechanisms that can lead to
stress-potentiated drinking and to discover methods to block or reverse these changes. Because the dynorphin
system plays a prominent role in stress and ethanol-related behaviors, and dynorphin is present in a major subset
of NAc neurons, Aim 1 will test the hypothesis that NAc G9a’s effects on stress-potentiated ethanol drinking are
mediated through dynorphin-positive neurons (NAcDyn+). This hypothesis will be tested by using a novel Cre-
dependent shRNA viral vector and a novel Cre-dependent G9a over-expression viral vector in both dynorphin-
Cre mice and enkephalin-Cre mice. Specifically, G9a will be knocked down or overexpressed in these NAc
neuronal subsets, and the effects of G9a will be tested on two different forms of stress-potentiated drinking. We
will also examine the NAc subregions involved in G9a’s effects. Next, Aim 2 will test the hypothesis that the
mechanism underlying G9a’s effects on stress-potentiated ethanol drinking involve NAc intrinsic excitability by
altering the expression of a specific potassium (K+) channel subunit. The hypothesis for Aim 2 is that the effects
of G9a on this K+ channel subunit reduces stress-potentiated drinking. We will directly test G9a’s ability to
regulate this K+ channel subunit with a CRISPR-fused G9a, and we will test G9a’s effects on transcription and
chromatin state using single nuclei multiomics. Finally, Aim 3 will test the hypothesis that the effects of NAc G9a
on stress-potentiated ethanol drinking are mediated via changes in activity in the bed nucleus of the stria
terminalis (BNST). Based on preliminary c-Fos data, Aim 3 will test the hypothesis that endogenous NAc G9a
increases stress-potentiated drinking by decreasing the activity of the BNST. Studies in this aim will first
determine which BNST subregions are influenced by alcohol, stress, and NAc G9a using immunolabeling-
enabled three-dimensional imaging of solvent-cleared organs (iDISCO). Next, the project will determine if these
c-Fos+ BNST neurons are sufficient to modulate stress-potentiated ethanol drinking by using Targeted
Recombination in Active Population (c-Fos-TRAP) mice combined with Cre-dependent expression of designer
receptors exclusively activated by designer drugs (DREADDs) in BNST ensembles. Finally, this Aim will directly
test the NAc to BNST pathway with a combination of a retrograde virus that expresses cre and a cre-dependent
shG9a virus. The central hypothesis is that NAc G9a increases stress-potentiate
Istituzione: LOUISIANA STATE UNIV A&M COL BATON ROUGE
PI: Ethan Michael Anderson
Progetto: 5R01AA031007-02
Settori: National Institute on Alcohol Abuse and Alcoholism
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