One concept to explain the etiology of ethanol-related neurological disorders is that ethanol alters the normal function of fast neurotransmission receptor systems in the brain. These receptors include the neuronal nicotine-activated receptors (nAChR) and glutamate-activated receptors (GluR). Understanding the mechanism(s) of ethanol's specific effects on these receptors is crucial to interrupting the habituating behavior and enhanced susceptibility to neurotoxicity that correlates with abuse of this compound. The intent of this RFA is to generate and distribute antibodies to proteins targeted by alcohol. This is the theme of our proposal and the interactive proposal submitted by Dr. Ronald Lukas of the Barrow Institute. We propose two Specific Aims. First, we will generate polyclonal and monoclonal antibodies to nAChR and GluR subunits and define their subunit specificity and their most useful application for immuno-detection of these proteins. Bacterially-produced fusion proteins or fusion-peptides will be used as immunogens, and the resulting antibodies will be screened and tested using a combination of Western blot analysis, immunoprecipitation, and immunohistochemistry. Major immunogenic regions will be defined by epitope mapping of specific immunoreactivities from the polyclonal antisera and this information used as a """"""""road map"""""""" to generate appropriate mouse monoclonal antibodies in amounts sufficient for distribution to other investigators. Second, we will use these antibodies for immunohistochemical analyses. Specifically, our experiments will test the hypothesis: Ethanol sensitive mice (LS or 'long-sleep') differ relative to ethanol insensitive mice (SS or 'short-sleep') in the expression and/or distribution of GluR and/or nAChR subunits normally or subsequent to chronic ethanol ingestion.

Agency
National Institute of Health (NIH)
Institute
National Institute on Alcohol Abuse and Alcoholism (NIAAA)
Type
Research Project (R01)
Project #
5R01AA011418-02
Application #
2516863
Study Section
Special Emphasis Panel (SRCA)
Project Start
1996-09-30
Project End
2000-08-31
Budget Start
1997-09-01
Budget End
1998-08-31
Support Year
2
Fiscal Year
1997
Total Cost
Indirect Cost
Name
University of Utah
Department
Biochemistry
Type
Schools of Medicine
DUNS #
City
Salt Lake City
State
UT
Country
United States
Zip Code
84112
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Gahring, L C; Carlson, N G; Wieggel, W A et al. (1999) Alcohol blocks TNFalpha but not other cytokine-mediated neuroprotection to NMDA. Alcohol Clin Exp Res 23:1571-9
Carlson, N G; Wieggel, W A; Chen, J et al. (1999) Inflammatory cytokines IL-1 alpha, IL-1 beta, IL-6, and TNF-alpha impart neuroprotection to an excitotoxin through distinct pathways. J Immunol 163:3963-8
Carlson, N G; Bacchi, A; Rogers, S W et al. (1998) Nicotine blocks TNF-alpha-mediated neuroprotection to NMDA by an alpha-bungarotoxin-sensitive pathway. J Neurobiol 35:29-36
Gahring, L C; Carlson, N G; Rogers, S W (1998) Antibodies prepared to neuronal glutamate receptor subunit3 bind IFNalpha-receptors: implications for an autoimmune process. Autoimmunity 28:243-8
Carlson, N G; Gahring, L C; Twyman, R E et al. (1997) Identification of amino acids in the glutamate receptor, GluR3, important for antibody-binding and receptor-specific activation. J Biol Chem 272:11295-301