The focus of this project continues to be transgenic dissection of the putative molecular targets of ethanol to explain the neurophysiological basis of action. The central hypothesis is that the type A gamma-aminobutyric acid receptor (GABA-A receptor) is a critical component of the mechanism of action that contributes to the behavioral response to ethanol. Mutants have been/ or will be created that harbor specific alterations in genes intimately involved in the neurochemical response to GABA. The mice are being/ will be analyzed with a battery of tests that span the molecular, cellular and behavioral levels; such a multi-level approach will allow responses to be properly attributed to a site of ethanol action. Specifically, mouse lines that ubiquitously lack either the beta 3, gamma 2L or delta subunits of the GABA-A receptor have already been produced; the electrophysiologic and behavioral responses of these animals to ethanol is being investigated. Two new mouse lines are being created (and will be analyzed) that harbor tissue-specific knockouts of the gamma isoform of protein kinase C; this locus has been demonstrated to be critical for behavioral responses to ethanol. The strategic advantages inherent in this conditional knockout approach are that untoward developmental effects of the genetic alteration are avoided, and the technique allows one to map the neuroanatomic location(s) that contribute to the behavioral phenotype observed. Three additional novel mouse lines are being created and will be analyzed; these are: (a) a ubiquitous knockout of the alpha1 subunit of the GABA-A receptor, (b) a transgene that expresses an Ala (291) Trp point mutation in the alpha1 subunit of the GABA-A receptor; the investigators have demonstrated electrophysiologically that this mutation abolishes ethanol enhancement of GABA action; a knockin mutation of the same Ala (291) Trp mutation that replaces the endogenous alpha1 gene. Together, these studies provide an integrated approach toward understanding how ethanol exerts its effects on the central nervous system.

Agency
National Institute of Health (NIH)
Institute
National Institute on Alcohol Abuse and Alcoholism (NIAAA)
Type
Research Project (R01)
Project #
5R01AA010422-07
Application #
6488800
Study Section
Special Emphasis Panel (ZRG4-ALTX-4 (02))
Program Officer
Neuhold, Lisa
Project Start
1995-08-01
Project End
2003-12-31
Budget Start
2002-01-01
Budget End
2002-12-31
Support Year
7
Fiscal Year
2002
Total Cost
$305,544
Indirect Cost
Name
University of Pittsburgh
Department
Anesthesiology
Type
Schools of Medicine
DUNS #
053785812
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213
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