Influence of Aging on Pathogenic Alpha-Synuclein Strains and Transmission Mechanism

《老龄化对致病性α-突触核蛋白毒株的影响及其传播机制》项目综述： 除了α-突触核蛋白的传递机制外，不同菌株的α-突触核蛋白的作用也是引人注目的 明白了。了解衰老对共核病发病机制的影响，包括 帕金森病(PD)、帕金森病伴痴呆(PDD)、路易体痴呆(DLB)和多系统疾病 萎缩症(MSA)是开发更有效的对症治疗的关键。在这份K01提案中，有三个 为了理解衰老的影响，提出了具体的目标：(1)产生和表征 从PD/PDD/DLB/MSA患者脑组织/脑脊液中提取致病毒株特异性α-SYN 纵向和横向老化因素。(2)了解年龄对区别性的影响 来自PD/PDD/DLB/MSA的致病α-SYN菌株的体内外。(三)揭开传播的面纱 与年龄相关的独特致病α-突触核蛋白的机制。实现3个具体目标，6个方法/步骤 需要：(I)胡安·特隆科索博士和利亚娜·罗森塔尔博士将为应聘者提供处理方面的培训 人脑组织/脑脊液。Ted博士和Valina Dawson博士以及Mark Mattson博士将使用 以脑组织/脑脊液中错折叠的α-突触核蛋白为模板，以蛋白质错折叠为环 扩增(PMCA)技术，以产生不同的α-突触核蛋白菌株。(Ii)扫描隧道显微镜 (扫描隧道显微镜)将用于成像不同的α-突触核蛋白菌株的分子结构，并区分 如果出现了潜在的问题，我们将向陈旺医生提供替代培训。 (Iii)原子力显微镜将用于观察不同α-突触核蛋白的组装形态 菌株。董明东博士将提供有关研究纳米机械性能和动力学的培训 不同α-突触核蛋白菌株的生长特性。(Iv)射击实验的电生理学研究将- 安东内洛·邦奇博士的训练。这项训练研究是为了了解固有的和突触的属性 受不同的α-突触核蛋白菌株的影响。(V)活体显微镜将由林大庭博士亲自动手培训。 这项训练包括执行所有必要的外科手术，将梯度折射率(GRIN)晶状体植入 黑质和双光子显微镜。这项研究将使候选人能够研究多巴胺神经元。 纹状体区立体定向注射不同α-突触核蛋白株对环路功能障碍的影响 评估变速箱。(Vi)淋巴细胞激活基因3(LAG3)已被鉴定为α-突触核蛋白 因此，LAG3是否可以介导PfF的传递是值得探讨的问题。 不同的α-突触核蛋白菌株。最重要的是，这个项目提出的是发展一个独立的研究实验室 有能力理解错误折叠结构上的淀粉样蛋白的不同菌株，传输和 神经退行性疾病的毒性，并成功地制定了一系列研究计划 争夺资金。

“Influence of Aging on Pathogenic α-Synuclein Strains and Transmission Mechanism” Project Summary: Besides α-synuclein transmission mechanism, the role of distinct strains of α-synuclein is compelling to be understood. Understanding the influence of aging on the pathogenesis of synucleinopathies, including Parkinson's disease (PD), PD with dementia (PDD), dementia with Lewy body (DLB) and multiple system atrophy (MSA), is crucial for developing more effective symptomatic therapies. In this K01 proposal, three specific aims are proposed for understanding the influence of aging: (1) To generate and characterize pathogenic strain-specific α-syn from brain tissue/CSF of patients with PD/PDD/DLB/MSA to controls by the factor of aging longitudinally and cross-sectionally. (2) To understand the influence of age on distinct pathogenic α-syn strains from PD/PDD/DLB/MSA in vitro and in vivo. (3) To uncover the transmission mechanism of age-related distinct pathogenic α-synuclein. To achieve the 3 specific aims, 6 methods/steps are required: (i) Dr. Juan Troncoso and Dr. Liana Rosenthal will provide the training to candidate on handling human brain tissue/CSF. Drs. Ted and Valina Dawson, and Dr. Mark Mattson will train the candidate using the misfolded α-synuclein in brain tissue/cerebrospinal fluid (CSF) as templates, and with protein misfolding cyclic amplification (PMCA) technique, to generate distinct α-synuclein strains. (ii) Scanning tunneling microscopy (STM) will be applied for imaging molecular structures of distinct α-synuclein strains and distinguish the differences, and the alternative training will be available from Dr. Chen Wang if the potential problems appear. (iii) Atomic force microscopy (AFM) will be applied for observing assembly morphologies of distinct α-synuclein strains. Dr. Mingdong Dong will provide the training on studying nano-mechanical properties and dynamic growth features of distinct α-synuclein strains. (iv) Electrophysiology study on firing experiment will be hands- on training from Dr. Antonello Bonci. This training study is to understand the intrinsic and synaptic properties affected by distinct α-synuclein strains. (v) In vivo microscopy will be hands-on training from Dr. Da-Ting Lin. This training includes performing all necessary surgical procedure to insert gradient index (GRIN) lens into substantia nigra and two-photon microscopy. This study will allow candidate to study the dopamine neuronal circuit dysfunction affected by stereotaxically injected with distinct α-synuclein strains in striatum region for evaluation the transmission. (vi) Lymphocyte-activation gene 3 (LAG3) has been identified as α-synuclein preformed fibrils (PFF) receptor, so it is worth to explore whether LAG3 can mediate the transmission of distinct α-synuclein strains. Above all, this project proposed is to develop an independent research laboratory equipped to understand the distinct strains of amyloid proteins on misfolded structures, the transmission and the toxicity in neurodegenerative disorders, and to develop a programmatic line of research by successfully competing for funding.