Perineuronal nets as a barrier for a-synuclein spreading

神经周围网作为α-突触核蛋白传播的屏障

• 批准号: 2739947
• 金额: --
• 依托单位: University of Leeds
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2022
• 资助国家: 英国
• 起止时间: 2022 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2739947
• 关键词: Perineuronal; nets; barrier; synuclein; spreading

Project Summary:Parkinson's disease (PD) is a progressive neurodegenerative disorder which is characterised by the presence of Lewy bodies, made up of aggregated misfolded a-synuclein (a-syn) amongst other proteins, in neurones causing cell death. a-syn can be transferred between neurones leading to Lewy body spread. a-syn misfolding and deposition also contributes to other synucleinopathies. Perineuronal nets (PNNs) are a type of extracellular matrix (ECM) found on the surface of neurones. PNNs modulate synapse formation, control neuroplasticity, act as a protective layer against toxic proteins and oxidative stress, serve as an ion reservoir and sequester molecules on neurones. PNNs are made up of hyaluronan (a glycosaminoglycan), chondroitin sulphate proteoglycans, HAPLNs (link proteins) and Tenascin-R (a glycoprotein). Current data shows that wildtype a-syn binds to chondroitin sulphates in rat neuronal cultures. Additionally, characterisation in human brains shows that PNN neurones are spared from Lewy pathology, whilst non-PNN neurones are not.The aims of this project are to (1), characterise the different a-syn species and their structures, retrieved from human PD samples. a-syn isolated from tissue samples will be analysed using fluorescent probe labelling, atomic force microscope and/or cryo-electron microscopy. (2), Determine the interactions between chondroitin sulphates, PNNs and different a-syn species identified in (1). This will be determined through expressing and purifying proteins, oligomer/ fibril preparation and using biophysical techniques to characterise these molecular interactions. (3), Determine the impact of the interactions between a-syn, chondroitin sulphates, PNNs on: a-syn internalisation and seeding, in addition to the impact on neuronal morphology and physiology. This will be determined using neuronal culture, immunocytochemistry, confocal microscopy and quantification, and electrophysiology.This research will improve our understanding of the fundamental molecular mechanisms by which a-syn interacts with PNNs and how PNNs may offer protection to neurones against a-syn related pathologies.

项目摘要：帕金森病(PD)是一种进行性神经退行性疾病，其特征是神经元中存在路易小体，由聚集的错误折叠的α-突触核蛋白(a-syn)和其他蛋白质组成，导致细胞死亡。A-syn可以在神经元之间转移，导致路易体扩散。A-SYN的错误折叠和沉积也会导致其他联核病。神经周围神经网络(PNNS)是一种存在于神经元表面的细胞外基质(ECM)。PNNS调节突触的形成，控制神经可塑性，作为有毒蛋白和氧化应激的保护层，作为神经元上的离子储存库和隔离分子。PNNS由透明质酸(一种糖胺聚糖)、硫酸软骨素蛋白多糖、HAPLN(连接蛋白)和Tenascin-R(一种糖蛋白)组成。目前的数据显示，在大鼠神经元培养中，野生型a-syn与硫酸软骨素结合。此外，在人脑中的特征表明PNN神经元不受路易病的影响，而非PNN神经元则不能。本项目的目的是(1)表征从人类PD样本中检索到的不同的a-syn种类及其结构。从组织样本中分离出的A-SYN将使用荧光探针标记、原子力显微镜和/或冷冻电子显微镜进行分析。(2)确定硫酸软骨素、PNNS与(1)中鉴定的不同a-syn物种之间的相互作用。这将通过表达和纯化蛋白质、低聚物/原纤维制备和使用生物物理技术来表征这些分子相互作用来确定。(3)确定a-syn、硫酸软骨素、PNNS之间的相互作用对a-syn内化和播种的影响，以及对神经元形态和生理的影响。这项研究将通过神经元培养、免疫细胞化学、共聚焦显微镜和定量以及电生理学来确定。这项研究将加深我们对a-syn与PNNS相互作用的基本分子机制的理解，以及PNNS如何为神经元提供保护以对抗a-syn相关的病理。