GLUTAMATE/NICOTINIC RECEPTOR ANTIBODIES AND ALCOHOL

谷氨酸/烟碱受体抗体和酒精

• 批准号: 2516863
• 负责人: LORISE C GAHRING
• 金额: $ 9.91万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-09-30 至 2000-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2516863
• 关键词: alcoholism /alcohol abuse; antibody; antibody specificity; biomedical resource; chimeric proteins; enzyme linked immunosorbent assay; epitope mapping; ethanol; glutamate receptor; immunocytochemistry; immunoprecipitation; laboratory mouse; laboratory rabbit; monoclonal antibody; nicotinic receptors; peptide chemical synthesis; receptor sensitivity; western blottings

DESCRIPTION (APPLICANT'S ABSTRACT): 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.

描述（申请人的摘要）：解释病因的一个概念 与乙醇相关的神经系统疾病是乙醇改变正常 大脑中快速神经传递受体系统的功能。 这些 受体包括神经元尼古丁激活的受体（NACHR）和 谷氨酸激活受体（GLUR）。 了解 乙醇对这些受体的特定影响对于中断 习惯行为和增强对神经毒性的敏感性 与滥用该化合物有关。 此RFA的目的是 生成并分配到酒精靶向的蛋白质的抗体。 这是 我们提案的主题和Dr. Dr.提交的互动提案的主题 巴罗研究所的罗纳德·卢卡斯（Ronald Lukas）。 我们提出了两个具体目标。 首先，我们将产生多克隆和 对NACHR和GLUR亚基的单克隆抗体并定义其亚基 特异性及其在这些免疫检测中最有用的应用 蛋白质。 细菌生产的融合蛋白或融合肽将是 用作免疫原子，将筛选和测试所得的抗体 结合蛋白质印迹分析，免疫沉淀和 免疫组织化学。 主要免疫原性将由表位定义 从多克隆抗血清中绘制特定的免疫反应性，这 信息用作“路线图”来生成适当的鼠标单克隆 足以分配给其他研究人员的抗体。 其次，我们将使用这些抗体进行免疫组织化学分析。 具体而言，我们的实验将检验以下假设：乙醇敏感 相对于乙醇不敏感的小鼠（SS或SS或 格鲁尔和/或nACHR的表达和/或分布中的“短腿”） 亚基通常或之后摄入慢性乙醇。