GABA-A Receptor Subunit and Protein Kinase Genes

Sikela, James M.

Abstract

Recently evidence has been obtained implicating several GABAA receptor and protein kinase genes as potentially important for the actions of ethanol at the GABAA receptor. For example, the gamma21 subunit of the receptor has been shown to be necessary for ethanol to exert its potentiating effect on the GABAA receptor. In addition, it appears that phosphorylation of this subunit, presumably by protein kinase C, is required to generate this effect. Therefore, the objective of this component will be to use a number of specific molecular biological approaches to determine whether there are changes in the sequence and/or expression in these genes in lines of mice that exhibit differential responsiveness to alcohol. Changes which influence the amino acid sequence of these genes will be sought by direct DNA sequencing of PCR- amplified DNA corresponding to the protein coding regions of selected GABAA receptor and protein kinase genes from lines of mice known to genetically differ in ethanol sensitivity, dependence, and preference. Changes in the expression of these genes in specific brain regions will be investigated by Northern analysis, in situ hybridization, Western analysis and quantitative receptor autoradiography. When changes in the structure and/or expression of any of these genes are identified, the extent to which a specific change correlates with a particular alcohol- related phenotype will be determined. Molecular genetic differences found to exhibit a significant correlation with an alcohol-related phenotype will provide a candidate gene (or genes) for genetic predisposition to alcoholism which could then be tested in human genetic studies.

Funding Agency

• Agency: National Institute of Health (NIH)
• Institute: National Institute on Alcohol Abuse and Alcoholism (NIAAA)
• Type: Specialized Center (P50)
• Project #: 5P50AA003527-19
• Application #: 5204201
• Support Year: 19
• Fiscal Year: 1996

Publications

Bennett, B; Carosone-Link, P; Beeson, M et al. (2008) Genetic dissection of quantitative trait locus for ethanol sensitivity in long- and short-sleep mice. Genes Brain Behav 7:659-68

Vasiliou, Vasilis; Ziegler, Thomas L; Bludeau, Pequita et al. (2006) CYP2E1 and catalase influence ethanol sensitivity in the central nervous system. Pharmacogenet Genomics 16:51-8

Bowers, Barbara J; Miyamoto-Ditmon, Jill; Wehner, Jeanne M (2006) Regulation of 5-HT2A/C receptors and DOI-induced behaviors by protein kinase Cgamma. Pharmacol Biochem Behav 85:441-7

Zimatkin, Sergey M; Pronko, Sergey P; Vasiliou, Vasilis et al. (2006) Enzymatic mechanisms of ethanol oxidation in the brain. Alcohol Clin Exp Res 30:1500-5

Radcliffe, Richard A; Bludeau, Pequita; Asperi, William et al. (2006) Confirmation of quantitative trait loci for ethanol sensitivity and neurotensin receptor density in crosses derived from the inbred high and low alcohol sensitive selectively bred rat lines. Psychopharmacology (Berl) 188:343-54

Smith, Amy M; Bowers, Barbara J; Radcliffe, Richard A et al. (2006) Microarray analysis of the effects of a gamma-protein kinase C null mutation on gene expression in striatum: a role for transthyretin in mutant phenotypes. Behav Genet 36:869-81

Bowers, Barbara J; Radcliffe, Richard A; Smith, Amy M et al. (2006) Microarray analysis identifies cerebellar genes sensitive to chronic ethanol treatment in PKCgamma mice. Alcohol 40:19-33

Haughey, Heather M; Kaiser, Alan L; Johnson, Thomas E et al. (2005) Norepinephrine transporter: a candidate gene for initial ethanol sensitivity in inbred long-sleep and short-sleep mice. Alcohol Clin Exp Res 29:1759-68

Wu, Peter H; Poelchen, Wolfgang; Proctor, William R (2005) Differential GABAB Receptor Modulation of Ethanol Effects on GABA(A) synaptic activity in hippocampal CA1 neurons. J Pharmacol Exp Ther 312:1082-9

Quertemont, Etienne; Eriksson, C J Peter; Zimatkin, Sergey M et al. (2005) Is ethanol a pro-drug? Acetaldehyde contribution to brain ethanol effects. Alcohol Clin Exp Res 29:1514-21