The goal of this project is to systematically delineate the effects of chronic intermittent ethanol (CIE) exposure on GABAergic neuroactive steroids and their regulation in limbic brain areas after stress challenge. Both neuronal and extracellular neuroactive steroid levels will be examined by immunohistochemistry as well as gas chromatography-mass spectroscopy (GC-MS) analysis of microdialysates and/or tissue specimens, allowing investigation of limbic brain regions for the first time. We propose to test the hypotheses that ethanol exposure dysregulates basal or stress-induced levels of these steroids and these changes are related to ethanol consumption. We will further explore genetic contributions to adaptations in neurosteroid levels or responses.
The first aim will determine the effects of CIE on neuronal vs. extracellular 3a,5a-THP levels in limbic/reward brain areas, including the prefrontal cortex, nucleus accumbens, amygdala, and bed nucleus of the stria terminalis of C57BL/6J mice.
Aim 2 will delineate the effect of acute stress challenge on neuronal vs. extracellular 3a,5a-THP in limbic brain areas and the effect of stress challenge following five cycles of CIE in C57BL/6J mice.
Aim 3 will determine if basal or CIE-related neuroactive steroid levels in limbic/reward brain areas are correlated with ethanol preference drinking across BXD mouse strains.
Aim 4 will extend and compare our studies on neuroactive steroid adaptations to CIE in mice to primates. We will delineate the effects of prolonged ethanol drinking on 3a,5a-THP levels in limbic brain areas of rhesus monkeys. Our results will be integrated with changes in neuronal function, neurochemistry, gene expression and ethanol drinking behavior studies by our collaborators under the same experimental conditions. These studies will elucidate the effects of ethanol on GABAergic neuroactive steroids in limbic regions of brain and provide novel insights into the role of basal and stress-induced neuroactive steroids in various allostatic adaptations to chronic intermittent ethanol exposure.

Public Health Relevance

The mechanisms that underlie the propensity to drink heavily and the elevation of drinking after chronic exposure are unknown. We propose that neuronal or extracellular changes in neuroactive steroids in brain regions that regulate addiction and ethanol consumption will impact neuronal function, neurochemistry, gene expression and alcohol drinking behavior. This work may lead to new therapeutics for alcoholism.

Agency
National Institute of Health (NIH)
Institute
National Institute on Alcohol Abuse and Alcoholism (NIAAA)
Type
Research Project--Cooperative Agreements (U01)
Project #
5U01AA020935-03
Application #
8606724
Study Section
Special Emphasis Panel (ZAA1-DD (51))
Program Officer
Liu, Qi-Ying
Project Start
2012-02-10
Project End
2017-01-31
Budget Start
2014-02-01
Budget End
2015-01-31
Support Year
3
Fiscal Year
2014
Total Cost
$196,642
Indirect Cost
$62,576
Name
University of North Carolina Chapel Hill
Department
Psychiatry
Type
Schools of Medicine
DUNS #
608195277
City
Chapel Hill
State
NC
Country
United States
Zip Code
27599
Maldonado-Devincci, Antoniette M; Cook, Jason B; O'Buckley, Todd K et al. (2014) Chronic intermittent ethanol exposure and withdrawal alters (3?,5?)-3-hydroxy-pregnan-20-one immunostaining in cortical and limbic brain regions of C57BL/6J mice. Alcohol Clin Exp Res 38:2561-71
Porcu, Patrizia; Morrow, A Leslie (2014) Divergent neuroactive steroid responses to stress and ethanol in rat and mouse strains: relevance for human studies. Psychopharmacology (Berl) 231:3257-72
Cook, Jason B; Werner, David F; Maldonado-Devincci, Antoniette M et al. (2014) Overexpression of the steroidogenic enzyme cytochrome P450 side chain cleavage in the ventral tegmental area increases 3?,5?-THP and reduces long-term operant ethanol self-administration. J Neurosci 34:5824-34
Maldonado-Devincci, Antoniette M; Beattie, Matthew C; Morrow, Danielle H et al. (2014) Reduction of circulating and selective limbic brain levels of (3?,5?)-3-hydroxy-pregnan-20-one (3?,5?-THP) following forced swim stress in C57BL/6J mice. Psychopharmacology (Berl) 231:3281-92
Fish, Eric W; Whitman, Buddy J; DiBerto, Jeff F et al. (2014) Effects of the neuroactive steroid allopregnanolone on intracranial self-stimulation in C57BL/6J mice. Psychopharmacology (Berl) 231:3415-23
Morrow, A Leslie (2014) Special issue in memory of Robert H. Purdy. Psychopharmacology (Berl) 231:3241-2
Porcu, Patrizia; Locci, Andrea; Santoru, Francesca et al. (2014) Failure of acute ethanol administration to alter cerebrocortical and hippocampal allopregnanolone levels in C57BL/6J and DBA/2J mice. Alcohol Clin Exp Res 38:948-58