NEUROFILAMENT INVOLVEMENT IN LEWY BODY DISORDERS

神经丝参与路易体疾病

• 批准号: 2271298
• 负责人: WILLIAM D HILL
• 金额: $ 8.96万
• 依托单位: AUGUSTA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-05-01 至 1999-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2271298
• 关键词: Alzheimer's disease; Chordata; Parkinson's disease; RNase protection assay; antibody; cellular pathology; epitope mapping; in situ hybridization; inclusion body; monoclonal antibody; nervous system disorder; neurofilament; neurofilament proteins; phosphorylation; posttranslational modifications; protein biosynthesis; protein isoforms; protein purification; proteolysis

Lewy bodies are filamentous inclusions that occur in selected brainstem, limbic, and neocortical neurons in several disabling and dementing neurodegenerative diseases, including Parkinson's disease, diffuse Lewy body disease, and a subset of Alzheimer's disease. Neither the pathogenic events leading to Lewy body formation, nor the composition of Lewy bodies are known. The long-term objectives of this research are to identify the key components of Lewy bodies, to characterize both immediate pathogenic events leading to the formation of Lewy bodies as well as earlier or more primary pathological events and to clarify the relationship between Alzheimer's disease and Lewy body associated disorders. In this proposal a limited set of goals will be pursued that focus on the role of the neurofilament triplet proteins in the formation of Lewy body filaments and on their role as a target in the pathogenesis of these disorders. Neurofilament proteins are the focus of the application because they are currently the most convincing and consistent of the potential Lewy body filament molecules. The overall hypothesis of this application is that neurofilament metabolism is disrupted in Lewy body associated disorders such as Parkinson's disease, diffuse Lewy body disease, and subsets of Alzheimer's disease. The specific aims center on neurofilament involvement in these Lewy body associated diseases. These aims incorporate analysis of neurofilament metabolism in vulnerable neuron populations at the level of mRNA and protein expression, including post-translational modifications to neurofilament subunits. To accomplish this epitope mapping will be employed with a unique library of antibodies combined with confocal microscopy, quantitative in situ hybridization and ribonuclease protection assays. The specific aims also incorporate the isolation and characterization of Lewy bodies in order to directly determine if neurofilament subunits are components and if they are post-translationally modified. Novel strategies will be used for the isolation of Lewy bodies (affinity based) and the production of Lewy body specific antibodies (tolerization based). Additionally, this information will be compared between cortical and subcortical neuronal populations as well as across disease categories. While neurofilaments have been strongly implicated in the formation of Lewy body filaments, and alterations in neurofilament processing are suspected in Lewy body associated disorders, the question of their involvement has not been settled. The intention of this proposal is to move towards a clearer assessment of the role of neurofilaments in these disorders.

路易体是出现在特定脑干中的丝状包涵体， 边缘系统和新皮层神经元在几个残疾和痴呆 神经退行性疾病，包括帕金森氏病、弥漫性路易氏病 身体疾病和阿尔茨海默病的一个子集。致病的 导致路易体形成的事件，也不是路易体的组成 是已知的。 这项研究的长期目标是确定关键 路易体的组成部分，以表征直接致病性 导致路易体形成的事件以及更早或更早的事件 主要病理事件，并澄清之间的关系 阿尔茨海默病和路易体相关疾病。本提案中 将追求一套有限的目标，重点是 神经丝三联体蛋白在路易体丝形成中的作用， 在这些疾病的发病机制中作为靶点的作用。 神经丝蛋白是应用的焦点，因为它们是 目前最有说服力和一致的潜在路易体 细丝分子 本申请的总体假设是神经丝 代谢在路易体相关疾病中被破坏， 帕金森病、弥漫性路易体病和阿尔茨海默病的亚类 疾病具体的目标集中在神经丝参与这些 路易体相关疾病。这些目标包括分析 脆弱神经元群体中的神经丝代谢水平 mRNA和蛋白质表达，包括翻译后修饰， 神经丝亚单位为了完成该表位作图， 与结合共聚焦显微镜的独特抗体库一起使用， 显微镜、定量原位杂交和核糖核酸酶保护 测定。具体目标还包括隔离和 路易体的表征以直接确定是否 神经丝亚单位是组成部分， 修改.新策略将用于分离路易体 （基于亲和力）和路易体特异性抗体的产生 （基于容忍度）。此外，这些信息将被比较 皮质和皮质下神经元群体之间以及 疾病类别。 虽然神经丝强烈地参与了 路易体细丝，以及神经丝加工的改变， 怀疑路易体相关疾病，他们的问题， 参与尚未解决。这项建议的目的是 朝着更清楚地评估神经丝在这些过程中的作用迈进， 紊乱