GABA AND NMDA RECEPTOR ANTIBODIES FOR ALCOHOL RESEARCH

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DESCRIPTION (APPLICANT'S ABSTRACT): The acute and chronic effects of alcohol abuse on brain function are diverse and profound. Recent work has identified the NMDA and GABA-A receptors as sensitive targets of ethanol's action in the brain. However, considerable controversy exists concerning the mechanism(s) by which ethanol produces these effects. Much of this controversy is likely due to the fact that each of these two receptors has a wide variety of different subunit compositions which result in receptors with markedly different properties and ethanol sensitivities. Moreover, the effects of ethanol on these receptors are also thought to be dependent on phosphorylation of these receptor proteins. However, there are insufficient high quality antibodies readily available to researchers to permit the adequate characterization of receptor subunit expression, composition and phosphorylation and this is a major impediment to this area of research. The current proposal will use previously described (and successful) technologies as well as new methodologies to generate subunit and phosphosite specific antibodies for the various subunits of these two receptors. Specifically, fusion proteins and/or peptides representing subunit specific regions and/or phosphosites of the various proteins will be used as immunogens. The resultant antibodies will be purified and characterized in terms of specificity and suitability for western blot, immunoprecipitation and immunohistochemistry. Effective antibodies will then be used by the P.I. and Co-P.I. for studies of the effects of ethanol on these receptors. In addition, these antibodies will also be made available to the research community at large through vendors of research products.