MECHANISM OF BETA AMYLOID NEUROTOXICITY

β淀粉样蛋白神经毒性的机制

• 批准号: 3417265
• 负责人: Bruce A YANKNER
• 金额: $ 17.69万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-08-01 至 1995-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3417265
• 关键词: Alzheimer's disease; amyloid proteins; biological signal transduction; calmodulin dependent protein kinase; electron microscopy; enzyme complex; enzyme inhibitors; epitope mapping; gene induction /repression; hippocampus; immunocytochemistry; immunoprecipitation; laboratory rat; microtubules; neural degeneration; neurofibrillary tangles; phosphorylation; posttranslational modifications; protease inhibitor; protein sequence; protein structure function; receptor; receptor binding; synthetic peptide; tachykinin; tau proteins; tissue /cell culture; western blottings

Deposition of the beta amyloid protein in the brain is a pathological hallmark of Alzheimer's disease (AD). The goal of this proposal is to understand the molecular basis of recently defined neurodegenerative effects of beta amyloid. The minimal peptide structural requirements for beta amyloid neurotoxic activity will be determined and used to synthesize small active peptides for receptor binding studies in cultured hippocampal neurons. The newly identified serpin-enzyme complex (SEC) receptor that binds beta amyloid and substance P will be characterized in hippocampal neurons. To determine the role of the SEC receptor, natural and artificial SEC receptor ligands will be tested for effects on beta amyloid neurotoxic activity. The cellular effects of beta amyloid which follow binding involve changes in cytoskeletal integrity. The AD-associated A68 protein, an abnormally phosphorylated tau protein, is induced in hippocampal neurons by beta amyloid. Epitope mapping and direct sequencing will be performed to determine if induction of A68 in culture is due to the same post- translational modification of tau that occurs in the AD brain. The mechanism of A68 induction will be examined by determining if beta amyloid stimulates protein kinase activity. The role of A68 induction and cytoskeletal disruption in the neurodegenerative process will be assessed by electron microscopy and immunogold labeling of cytoskeletal proteins. Tachykinin peptides inhibit the neurotoxicity of beta amyloid in culture and in vivo in the adult rat brain. The mechanism of this effect will be initially examined by determining if it involves either the tachykinin NK1 receptor or the serpin receptor. The peptide structural requirements for inhibiting beta amyloid neurotoxicity will be determined using deletions, substitutions, and modifications of the substance P peptide. The studies outlined in this proposal will characterize a potential model system for the pathogenesis of AD and may lead to the rational design of new therapeutic interventions for AD.

β淀粉样蛋白在大脑中的沉积是一种病理 阿尔茨海默氏病（AD）的标志。 该提议的目的是 了解最近定义的神经退行性的分子基础 β-淀粉样蛋白的影响。 最小的肽结构要求 将确定β-淀粉样神经毒性活性并用于合成 小型活性肽用于培养海马的受体结合研究 神经元。 新鉴定的Serpin-酶复合物（SEC）受体 结合β淀粉样蛋白和物质P将在海马中进行表征 神经元。 为了确定SEC受体的作用，自然和人造 SEC受体配体将对β淀粉样神经毒性的影响进行测试 活动。 遵循结合的β-淀粉样β淀粉样蛋白的细胞效应涉及变化 在细胞骨架完整性中。 广告相关的A68蛋白，异常 磷酸化的tau蛋白是由β在海马神经元中诱导的 淀粉样蛋白。 表位映射和直接测序将进行 确定培养中A68的诱导是否是由于相同的后 在广告大脑中发生的Tau的翻译修饰。 这 通过确定β-淀粉样蛋白是否是否会检查A68诱导的机理 刺激蛋白激酶活性。 A68归纳和 将评估神经退行性过程中的细胞骨架破坏 通过电子显微镜和细胞骨架蛋白的免疫金标记。 旋转肽肽抑制培养基中β-淀粉样蛋白的神经毒性 和成年大鼠大脑中的体内。 这种效果的机制将是 最初通过确定是否涉及旋转金蛋白NK1来检查 受体或SERPIN受体。 肽结构要求 抑制β-淀粉样神经毒性将使用缺失确定 替代和对物质P肽的修饰。 研究 该提案中概述的概述将表征一个潜在的模型系统 AD的发病机理并可能导致新的新设计 AD的治疗干预措施。