NEUROFILAMENT INVOLVEMENT IN LEWY BODY DISORDERS

神经丝参与路易体疾病

• 批准号: 2416348
• 负责人: WILLIAM D HILL
• 金额: $ 9.7万
• 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/2416348
• 关键词: Alzheimer's disease; Chordata; Lewy body; Parkinson's disease; RNase protection assay; antibody; cellular pathology; epitope mapping; in situ hybridization; 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.

路易小体是出现在选定的脑干中的丝状包裹体， 几种致残和痴呆患者的边缘神经元和新皮质神经元 神经退行性疾病，包括帕金森氏病，弥漫性路易氏 躯体疾病，以及阿尔茨海默氏症的一个子集。既不是致病的 导致路易体形成的事件，也不是路易体的组成 都是已知的。 这项研究的长期目标是确定关键 路易小体的成分，以表征两种直接致病 导致路易体形成的事件以及更早或更早的事件 并阐明原发病理事件之间的关系 阿尔茨海默病和路易体相关疾病。在本建议书中 将追求一套有限的目标，重点放在 神经丝三联体蛋白在路易体细丝形成中的作用 关于它们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重点，因为它们是 目前最令人信服和一致的潜在路易体 细丝分子。 这一应用的总体假设是神经丝 路易体相关疾病的新陈代谢紊乱，如 帕金森氏病、弥漫性路易体病和阿尔茨海默病的亚型 疾病。具体的目标集中在神经丝参与这些 路易体相关疾病。这些目标结合了对 脆弱神经元群体中神经丝代谢水平的研究 MRNA和蛋白质的表达，包括翻译后修饰 神经丝亚单位。要完成这一表位映射，需要 使用独特的抗体库，结合共聚焦 显微镜、定量原位杂交和核糖核酸酶保护 化验。具体目标还包括孤立和 刘易体的表征，以便直接确定是否 神经丝亚单位是组成成分，如果它们是翻译后的 修改过的。将使用新的策略分离路易小体 (基于亲和力)和路易体特异性抗体的产生 (基于容忍度)。此外，还将比较这些信息 皮质和皮质下神经元群之间以及横跨 疾病类别。 而神经丝被强烈地牵涉到 路易体细丝，神经细丝加工中的变化是 怀疑与路易体相关的障碍，他们的问题 参与其中的问题尚未得到解决。这项提议的目的是 对神经丝在这些过程中的作用进行更清晰的评估 精神错乱。