High-content analysis of α-synuclein aggregation and cell death in a cellular model of Parkinson's disease

Background: Alpha-synuclein (alpha-SYN) aggregates represent a key feature of Parkinson's disease, but the exact relationship between alpha-SYN aggregation and neurodegeneration remains incompletely understood. Therefore, the availability of a cellular assay that allows medium-throughput analysis of alpha-SYN-linked pathology will be of great value for studying the aggregation process and for advancing alpha-SYN-based therapies. New method: Here we describe a high-content neuronal cell assay that simultaneously measures oxidative stress-induced alpha-SYN aggregation and apoptosis. Results: We optimized an automated and reproducible assay to quantify both alpha-SYN aggregation and cell death in human SH-SY5Y neuroblastoma cells. Comparison with existing methods: Quantification of alpha-SYN aggregates in cells has typically relied on manual imaging and counting or cell-free assays, which are time consuming and do not allow a concurrent analysis of cell viability. Our high-content analysis method for quantification of alpha-SYN aggregation allows simultaneous measurements of multiple cell parameters at a single-cell level in a fast, objective and automated manner. Conclusions: The presented analysis approach offers a rapid, objective and multiparametric approach for the screening of compounds and genes that might alter alpha-SYN aggregation and/or toxicity. (C) 2016 Published by Elsevier B.V.