Furman Jennifer L, Diamond Marc I
Center for Alzheimer's and Neurodegenerative Diseases, Dallas, TX, USA.
University of Texas-Southwestern Medical Center, 6000 Harry Hilned Blvd., NL 10.120K, Dallas, TX, 75390, USA.
Methods Mol Biol. 2017;1523:349-359. doi: 10.1007/978-1-4939-6598-4_23.
Transcellular propagation of protein aggregates-or seeds-is increasingly implicated as a mechanism for disease progression in many neurodegenerative disorders, including Alzheimer's disease and the related tauopathies. While neuropathology generally originates in one discrete brain region, pathology progresses as disease severity advances, often along discrete neural networks. The stereotypical spread of tau pathology suggests that cell-to-cell transfer of toxic protein aggregates could underlie disease progression, and recent studies implicate seeding as a proximal marker of disease, as compared to standard histological and biochemical analyses. Commonly used metrics for protein aggregation detection, however, lack sensitivity, are not quantitative, and/or undergo subjective classification. Here, we describe a FRET and flow cytometry cell-based assay that allows for rapid and quantitative detection of protein aggregates from human and rodent biological specimens.
蛋白质聚集体（或种子）的跨细胞传播越来越被认为是包括阿尔茨海默病和相关tau蛋白病在内的许多神经退行性疾病进展的一种机制。虽然神经病理学通常起源于一个离散的脑区，但随着疾病严重程度的增加，病理学往往沿着离散的神经网络进展。tau蛋白病理学的典型传播表明，有毒蛋白质聚集体的细胞间转移可能是疾病进展的基础，与标准组织学和生化分析相比，最近的研究表明种子形成是疾病的近端标志物。然而，常用的蛋白质聚集检测指标缺乏敏感性、不是定量的，和/或需要进行主观分类。在这里，我们描述了一种基于FRET和流式细胞术的细胞检测方法，该方法可以快速、定量地检测来自人和啮齿动物生物标本的蛋白质聚集体。
