Summary
This project aims to elucidate the epigenetic mechanisms, specifically involving G9a, that regulate stress-potentiated ethanol drinking and to discover methods to block or reverse these changes.
Epigenetic regulation of stress-potentiated ethanol drinking
US · IL
NIH
grant
awarded
#nih-5R01AA031007-02
What they want
The project will investigate three main aims: 1) Test the hypothesis that NAc G9a's effects on stress-potentiated ethanol drinking are mediated through dynorphin-positive neurons (NAcDyn+) using novel Cre-dependent viral vectors in dynorphin-Cre and enkephalin-Cre mice. 2) Test the hypothesis that the mechanism involves NAc intrinsic excitability by altering a specific potassium (K+) channel subunit, utilizing CRISPR-fused G9a and single nuclei multiomics. 3) Test the hypothesis that NAc G9a's effects are mediated via changes in activity in the bed nucleus of the stria terminalis (BNST), employing iDISCO, c-Fos-TRAP mice with DREADDs, and retrograde viral tracing.
Deliverables
- Elucidation of mechanisms leading to stress-potentiated drinking
- Discovery of methods to block or reverse stress-potentiated drinking changes
- Enhanced understanding of how stress potentiates alcohol drinking
Technical requirements
- Cre-dependent shRNA viral vector
- Cre-dependent G9a over-expression viral vector
- Dynorphin-Cre mice
- Enkephalin-Cre mice
- CRISPR-fused G9a
- Single nuclei multiomics
- Immunolabeling-enabled three-dimensional imaging of solvent-cleared organs (iDISCO)
- Targeted Recombination in Active Population (c-Fos-TRAP) mice
- Cre-dependent expression of designer receptors exclusively activated by designer drugs (DREADDs)
- Retrograde virus that expresses cre
