Kedia Shekhar, Fertan Emre, Wu Yunzhao, Zhang Yu P, Meisl Georg, Lam Jeff Y L, K Wiseman Frances, McEwan William A, Quaegebeur Annelies, Spillantini Maria Grazia, Danial John S H, Klenerman David
Yusuf Hamied Department of Chemistry, University of Cambridge Cambridge CB2 1EW, UK; UK Dementia Research Institute at University of Cambridge Cambridge CB2 0XY, UK.
Yusuf Hamied Department of Chemistry, University of Cambridge Cambridge CB2 1EW, UK; UK Dementia Research Institute at University of Cambridge Cambridge CB2 0XY, UK; Department of Clinical Neurosciences, University of Cambridge Cambridge, UK.
Cell Chem Biol. 2025 Feb 20;32(2):338-351.e4. doi: 10.1016/j.chembiol.2025.01.001. Epub 2025 Jan 24.
Synaptic dysfunction is a primary hallmark of both Alzheimer's and Parkinson's disease, leading to cognitive and behavioral decline. While alpha-synuclein, beta-amyloid, and tau are involved in the physiological functioning of synapses, their pathological aggregation has been linked to synaptopathology. The methodology for studying the small-soluble protein aggregates formed by these proteins is limited. Here we describe SynPull, a method combining single-molecule pull-down, super-resolution microscopy, and advanced computational analyses to characterize the protein aggregates in human and mouse synaptosomes. We show that AT8-positive tau aggregates are the predominant aggregate type in synaptosomes from postmortem Alzheimer's disease brain, although the aggregate size does not change in disease. Meanwhile, the relatively smaller amount of alpha-synuclein and beta-amyloid aggregates found in the synapses are larger than the extra-synaptic ones. Collectively, these results show the utility of SynPull to study pathological aggregates in neurodegeneration, elucidating the disease mechanisms causing synaptic dysfunction.
突触功能障碍是阿尔茨海默病和帕金森病的主要特征，会导致认知和行为能力下降。虽然α-突触核蛋白、β-淀粉样蛋白和tau蛋白参与突触的生理功能，但它们的病理性聚集与突触病变有关。研究由这些蛋白质形成的小可溶性蛋白质聚集体的方法有限。在此，我们描述了SynPull，这是一种结合单分子下拉、超分辨率显微镜和先进计算分析来表征人和小鼠突触小体中蛋白质聚集体的方法。我们发现，死后阿尔茨海默病大脑的突触小体中，AT8阳性tau聚集体是主要的聚集体类型，尽管疾病状态下聚集体大小没有变化。同时，在突触中发现的相对少量的α-突触核蛋白和β-淀粉样蛋白聚集体比突触外的聚集体更大。总体而言，这些结果表明SynPull在研究神经退行性疾病中的病理性聚集体方面具有实用性，有助于阐明导致突触功能障碍的疾病机制。
