GABA AND NMDA RECEPTOR ANTIBODIES FOR ALCOHOL RESEARCH

描述（申请人摘要）： 酒精滥用对大脑功能的影响是多样而深刻的。 最近的工作已经 确定NMDA和GABA-A受体为乙醇的敏感靶点， 大脑中的活动。 然而，在这方面存在着相当大的争议。 乙醇产生这些效应的机制。 这在很大程度 争议可能是由于这两种受体中的每一种都具有 产生受体的各种不同亚基组成 具有显著不同的性质和乙醇敏感性。 而且 乙醇对这些受体的影响也被认为取决于 这些受体蛋白的磷酸化。 然而， 高质量的抗体随时可供研究人员使用， 充分表征受体亚基表达、组成和 磷酸化，这是这一研究领域的主要障碍。 目前的建议将使用先前描述的（和成功的） 技术以及新的方法，以产生亚单位， 针对这两个亚基的磷酸化特异性抗体 受体。 具体地，表达以下的融合蛋白和/或肽： 各种蛋白质的亚基特异性区域和/或磷酸化位点将被 用作免疫原。 将所得抗体纯化， 其特征在于特异性和对蛋白质印迹的适用性， 免疫沉淀和免疫组织化学。 有效的抗体将 然后被私家侦探利用和Co-P.I.研究乙醇的影响 在这些受体上 此外，这些抗体也将被制成 通过研究供应商提供给整个研究社区 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.