Enteric Neuronal Aggregation of a-synuclein causes disruption in Colonic Neurotransmission and results in Colonic Dysmotility

α-突触核蛋白的肠道神经元聚集会导致结肠神经传递中断并导致结肠动力障碍

• 批准号: 10390438
• 负责人: Narayana Krishna Yelleswarapu
• 金额: $ 4.76万
• 依托单位: MICHIGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-16 至 2023-05-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10390438
• 关键词: Address; Animal Model; Bathing; Biological Assay; Braak' s hypothesis; Clinical; Colon; Confocal Microscopy; Constipation; Data; Dietary Intervention; Digestion; Distal; Dopaminergic Agents; Eating; Economic Burden; Ectopic Expression; Electrodes; Electrophysiology (science); Enteral; Enteric Nervous System; Gastrointestinal tract structure; Goals; Health; Health Professional; Immunohistochemistry; Impairment; Individual; Isometric Exercise; Knowledge; Label; Lead; Light; Link; Mediating; Migrating Myoelectric Complex; Motor; Motor Neurons; Mus; Myenteric Plexus; Neuraxis; Neurodegenerative Disorders; Neurons; Neurotransmitters; Organ; Output; Parkinson Disease; Pathologic; Pathologic Processes; Pathology; Patients; Pattern; Pharmacotherapy; Physical therapy; Point Mutation; Population; Presynaptic Terminals; Proteins; Quality of life; Recombinant adeno-associated virus (rAAV); Relaxation; Research; Role; Smooth Muscle; Smooth Muscle Myocytes; Symptoms; Synapses; Synaptic Transmission; Synaptic Vesicles; System; Testing; Time; Tissues; Transgenic Organisms; Weight; alpha synuclein; cell motility; cholinergic; cholinergic neuron; drug discovery; effective therapy; gastrointestinal; in vivo; laxative; motility disorder; motor deficit; motor impairment; mouse model; neuromuscular; neuromuscular transmission; neuronal circuitry; neurotransmission; new therapeutic target; novel; novel therapeutics; optogenetics; overexpression; retrograde transport; targeted treatment

Abstract Gastrointestinal (GI) complications are a significant clinical and economic burden in patients with Parkinson’s disease (PD). Specifically, constipation is responsible for the most drastic decrease in the quality of life for 24- 63% of PD patients. Yet, due to the lack of knowledge of the mechanisms underlying this symptom, there haven’t been successful or long-term therapies. It has been well established that aggregated forms of α- synuclein (α-syn), a presynaptic terminal protein identified to be the pathological marker in PD, is observed diversely through the enteric nervous system (ENS) of the GI tract in PD patients. α-syn aggregation within the ENS as well as constipation precede the motor deficits in PD by up to 20 years. Whether this protein is interfering with neurotransmission in the ENS is currently unknown. Our preliminary data in recombinant adeno-associated virus with A53T point mutation targeted to enteric neurons in the mouse model found reduced colonic propulsion suggesting that α-syn maybe disrupting cholinergic neurotransmission. The long- term objective of this project is to determine whether α-syn aggregation in the colonic myenteric plexus impairs ENS cholinergic neuromuscular and synaptic transmission causing abnormal colonic propulsion. To achieve this project objective, Aim 1 will determine the expression of ectopic α-syn in the myenteric neurons of the colon as this will provide us mechanistic data on the potential interactions between the protein and neurotransmitter systems. In Aim 2 we will use in vivo and ex vivo paradigms to determine whether α-syn overexpression disrupts propulsive colonic motility by addressing colonic propulsion and colonic contractions and relaxations. Aim 3 will use optogenetics to identify the effect of α-syn overexpression on cholinergic synaptic and neuromuscular transmission in the myenteric plexus. This proposed research will be the first to directly study cholinergic neurotransmission within the ENS in the presence of α-syn aggregates. Moreover, we aim to identify novel targets that may initiate new drug discoveries in treating constipation involved in PD.

摘要 胃肠道（GI）并发症是帕金森病患者的重要临床和经济负担 疾病（PD）。具体来说，便秘是造成24岁以下人群生活质量下降最严重的原因。 63%的PD患者。然而，由于缺乏对这种症状背后机制的了解， 还没有成功或长期的治疗方法。已经很好地确定，α- 突触核蛋白（α-syn）是一种突触前终末蛋白，被鉴定为PD的病理标志物 在PD患者中，通过胃肠道的肠神经系统（ENS）进行功能锻炼。α-syn聚集在 ENS以及便秘先于PD患者的运动缺陷长达20年。这种蛋白质是否 干扰ENS中的神经传递目前是未知的。我们在重组方面的初步数据 在小鼠模型中发现了靶向肠神经元的A53 T点突变腺相关病毒 降低结肠推进力，提示α-syn可能干扰胆碱能神经传递。很长的- 本项目的长期目标是确定α-syn聚集在结肠肌间 神经丛损害ENS胆碱能神经肌肉和突触传递，导致结肠异常 推进力为了实现这一项目目标，目标1将确定异位α-syn在 结肠肌间神经元，因为这将为我们提供关于 蛋白质和神经递质系统。在目标2中，我们将使用体内和离体范例来确定 α-syn过表达是否通过解决结肠推进和结肠收缩来破坏推进性结肠运动 收缩和舒张。目的3将利用光遗传学来鉴定α-syn过表达对 肌间神经丛中的胆碱能突触和神经肌肉传递。这项研究将是 第一个直接研究在α-syn聚集体存在下ENS内胆碱能神经传递。 此外，我们的目标是确定新的目标，可能会启动新的药物发现治疗便秘 参与PD。