GABA AND NMDA RECEPTOR ANTIBODIES FOR ALCOHOL RESEARCH

描述(申请人摘要)：急性和慢性影响 酒精滥用对大脑功能的影响是多样和深远的。最近的工作有 确定NMDA和GABA-A受体为乙醇的敏感靶点 大脑中的活动。然而，关于以下问题仍存在相当大的争议 乙醇产生这些效应的机制(S)。其中很大一部分 争议可能是因为这两个受体都有一个 导致受体的各种不同的亚基组成 具有明显不同的性质和乙醇敏感性。此外， 乙醇对这些受体的影响也被认为依赖于 这些受体蛋白的磷酸化。然而，没有足够的 研究人员随时可以获得高质量的抗体，以允许 充分表征受体亚单位的表达、组成和 磷酸化，这是这一研究领域的主要障碍。 目前的提案将使用前面描述的(并且成功) 技术以及新的方法来生成亚单位和 这两个亚基不同亚基的亚磷酸盐特异性抗体 感受器。具体而言，融合蛋白和/或代表 亚基特定区域和/或各种蛋白质的亚磷酸盐 用作免疫原。所产生的抗体将被提纯并 以蛋白质印迹的特异性和适合性为特征， 免疫沉淀和免疫组织化学。有效的抗体将 然后被P.I.和Co-P.I.用于研究乙醇的影响 在这些受体上。此外，这些抗体也将被制成 通过研究供应商向广大研究社区提供 产品。

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.