Alcohol abuse and alcoholism are serious health problems in the United States. However, the molecular bases of alcohol's actions and alcoholism are unknown. The overall goal of this proposal is to study the molecular mechanism(s) of action of alcohol at the neurotransmitter-gated ion channel level. The effects of ethanol on GABA-A and NMDA receptors, the major inhibitory and excitatory neurotransmitter-gated ion channels in the mammalian central nervous system, respectively, will be studied. The function of these ion channels is regulated by protein phosphorylation/dephosphorylation. This regulatory process is mediated by multiple kinases and phosphatases, and appears to be important for ethanol's actions. A combination of electrophysiological and biochemical techniques in cultured transfected cell lines is proposed to be used to study the role of protein phosphorylation in the ethanol-induced effects on these channels. Electropysiological experiments will determine the effect of ethanol on receptor function. Immunoprecipitation experiments in the same batch of cells will directly examine the phosphorylation state of GABA-A and NMDA receptor subunits in the presence of ethanol. If ethanol changes the phosphorylation state of receptor subunits, then phosphoamino acid and phosphopeptide analyses, and studies with kinase & phosphatase inhibitors, will determine which kinases or phosphatases are required for ethanol's actions. Correlation of electrophysiological and site-directed mutagenesis experiments will determine whether phosphorylation of specific amino acids in GABA-A AND NMDA receptor subunits is required for ethanol's actions. The findings of the proposed experiments will contribute toward a better understanding of the actions of ethanol in the central nervous system.

Agency
National Institute of Health (NIH)
Institute
National Institute on Alcohol Abuse and Alcoholism (NIAAA)
Type
Research Scientist Development Award - Research & Training (K01)
Project #
5K01AA000227-02
Application #
2457450
Study Section
Biochemistry, Physiology and Medicine Subcommittee (ALCB)
Project Start
1996-08-01
Project End
1998-07-31
Budget Start
1997-08-01
Budget End
1998-07-31
Support Year
2
Fiscal Year
1997
Total Cost
Indirect Cost
Name
University of Colorado Denver
Department
Pharmacology
Type
Schools of Medicine
DUNS #
065391526
City
Aurora
State
CO
Country
United States
Zip Code
80045
Cardoso, R A; Brozowski, S J; Chavez-Noriega, L E et al. (1999) Effects of ethanol on recombinant human neuronal nicotinic acetylcholine receptors expressed in Xenopus oocytes. J Pharmacol Exp Ther 289:774-80
Valenzuela, C F; Kazlauskas, A; Weiner, J L (1997) Roles of platelet-derived growth factor in the developing and mature nervous systems. Brain Res Brain Res Rev 24:77-89