Mechanisms of synaptic dysfunction in Parkinson's and other synuclein-linked dise

帕金森病和其他突触核蛋白相关疾病中突触功能障碍的机制

• 批准号: 8602862
• 负责人: Jennifer Rebecca Morgan
• 金额: $ 31.28万
• 依托单位: MARINE BIOLOGICAL LABORATORY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-01-15 至 2016-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8602862
• 关键词: ATP phosphohydrolase; Acute; Address; Affect; Affinity; Alzheimer' s Disease; Appearance; Binding; Biochemical; Biological Assay; Cell membrane; Clathrin; Cognitive deficits; Complement; Data; Defect; Dementia; Development; Disease; Endocytosis; Event; Functional disorder; Genetic; Goals; Human; In Vitro; Knowledge; Lampreys; Lead; Lewy Body Dementia; Link; Lipid Binding; Lipids; Maps; Masks; Measures; Mediating; Membrane Lipids; Modeling; Molecular; Molecular Target; Mutation; N-terminal; Nerve Degeneration; Neurodegenerative Disorders; Neuromuscular Diseases; Neurons; Parkinson Disease; Pathway interactions; Patients; Peptides; Phenotype; Phosphatidylinositols; Public Health; Reagent; Recycling; Research; Staging; Stroke; Surface Plasmon Resonance; Synapses; Synaptic Transmission; Synaptic Vesicles; Testing; Variant; Vesicle; Work; design; improved; in vivo; innovation; insight; mutant; nervous system disorder; novel; novel strategies; overexpression; presynaptic; prevent; research study; spinal cord and brain injury; synaptic function; synuclein; synucleinopathy; trafficking; uptake

ABSTRACT The long term goal of this project is to identify the cellular and molecular mechanisms that give rise to the synaptic defects in patients with Parkinson's disease (PD) and other related neurological disorders. The pathological hallmark of these diseases includes abnormal levels of ¿-synuclein at synapses and throughout the neuron. While it is generally agreed that synucleins participate in synaptic vesicle trafficking, the exact steps of the vesicle trafficking pathway that are perturbed by altered levels of synuclein remain unclear. Thus, at present, it is not possible to design targeted strategies for improving synaptic function in PD. Experiments proposed here take the first steps toward this by identifying the precise synaptic vesicle trafficking defects caused excess synuclein at synapses, the mechanisms giving rise to these defects, and new strategies for reversing them. The experiments take advantage of two model synapses that are ideally suited for studies of synaptic vesicle trafficking, using both acute and genetic perturbations. The combination of highly quantitative biochemical assays to measure synuclein interactions, design of reagents to perturb these interactions, and detailed ultrastructural analyses provides the best opportunity to identify the cellular and molecular mechanisms leading to synuclein-induced synaptic vesicle trafficking defects. In initial studies, excess wild type synuclein causes a loss of synaptic vesicles, increased cisternae, and altered clathrin-coated profiles, consistent with inhibiting clathrin-mediated synaptic vesicle recycling. Going forward, proposed experiments are aimed at identifying the cellular mechanisms by which excess wild type ¿-synuclein and PD-related mutations (e.g. A30P, E46K, A53T) cause vesicle trafficking defects (Aims 1 and 2). Experiments will also investigate how synuclein interactions with specific synaptic binding partners (e.g. PI(4,5)P2 and the uncoating ATPase) contribute to the synaptic vesicle trafficking defects, and targeted strategies for disrupting these interactions will be assessed as a possible means for reversing synaptic defects (Aim 2 and 3). The proposed experiments are innovative because they are the first to use a combination of quantitative biochemical binding assays, acute perturbations, controlled stimulation conditions, and detailed ultrastructural analyses to identify the precise synaptic vesicle trafficking defects caused by excess synuclein or its mutations, which is ideally suited for the overall goal. The experiments are significant because they represent the first steps toward understanding the mechanisms giving rise to the synaptic defects, and they provide possible targeted, molecular strategies for improving synaptic function. Thus, these studies have direct implications for slowing or halting the neurodegeneration, cognitive deficits, and dementia in PD and other synucleinopathies.

摘要 这个项目的长期目标是确定引起 帕金森氏病(PD)和其他相关神经疾病患者的突触缺陷。 这些疾病的病理特征包括突触和突触中突触核蛋白的异常水平 贯穿整个神经元。虽然普遍认为突触核蛋白参与突触小泡 运输，囊泡运输途径的确切步骤被改变的水平扰乱 突触核蛋白仍不清楚。因此，目前不可能制定有针对性的改进战略。 帕金森病的突触功能。这里提出的实验是通过识别 精确的突触囊泡运输缺陷导致突触处突触核蛋白过量，机制 导致这些缺陷的原因，以及扭转这些缺陷的新策略。这些实验利用了 两种模型突触非常适合研究突触囊泡运输，使用两种 和基因扰动。结合高定量生化分析进行检测 突触核蛋白相互作用，干扰这些相互作用的试剂的设计，以及详细的超微结构 分析提供了最好的机会来确定导致 突触核蛋白诱导的突触小泡运输缺陷。在最初的研究中，过量的野生型突触核蛋白 导致突触小泡丢失，脑池增多，并改变笼状蛋白包被的轮廓，符合 通过抑制网状蛋白介导的突触小泡循环。展望未来，拟议的实验是 目的是确定过量野生型突触核蛋白与帕金森病相关的细胞机制 突变(如A30P、E46K、A53T)导致囊泡运输缺陷(目标1和2)。实验将会 也研究突触核蛋白如何与特定的突触结合伙伴(例如PI(4，5)P2和 脱壳的ATPase)导致突触小泡运输缺陷，并针对 破坏这些相互作用将被评估为逆转突触缺陷(AIM)的可能手段 2和3)。拟议中的实验具有创新性，因为它们是第一个使用 定量生化结合分析，急性扰动，受控刺激条件，以及 详细的超微结构分析以确定准确的突触小泡运输缺陷 过量的突触核蛋白或其突变，这是最适合总体目标的。这些实验是 意义重大，因为它们代表了理解导致 突触缺陷，它们为改善突触提供了可能的靶向分子策略 功能。因此，这些研究对减缓或阻止神经退行性变具有直接意义， 认知缺陷、帕金森病和其他联核症中的痴呆症。