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

被引：14

作者：

Macchi, Francesca ^{[1
]}

Deleersnijder, Angelique ^{[1
]}

Van den Haute, Chris ^{[1
,4
]}

Munck, Sebastian ^{[2
]}

Pottel, Hans ^{[3
]}

Michiels, Annelies ^{[1
,4
]}

Debyser, Zeger ^{[5
]}

Gerard, Melanie ^{[1
,6
]}

Baekelandt, Veerle ^{[1
]}

机构：

[1] Katholieke Univ Leuven, Lab Neurobiol & Gene Therapy, Kapucijnenvoer 33, B-3000 Leuven, Flanders, Belgium

[2] Flanders Interuniv Inst Biotechnol, Dept Human Genet, KU Leuven, Kapucijnenvoer 33, B-3000 Leuven, Flanders, Belgium

[3] KU Leuven Campus Kulak Kortrijk, Publ Hlth & Primary Care, Interdisciplinary Res Facil Life Sci, Etienne Sabbelaan 53, B-8500 Kortrijk, Belgium

[4] Katholieke Univ Leuven, Leuven Viral Vector Core, B-3000 Leuven, Flanders, Belgium

[5] Katholieke Univ Leuven, Lab Mol Virol & Gene Therapy, Dept Pharmaceut & Pharmacol Sci, B-3000 Leuven, Flanders, Belgium

[6] KU Leuven campus Kulak Kortrijk, Biochem Lab, Interdisciplinary Res Facil Life Sci, Etienne Sabbelaan 53, B-8500 Kortrijk, Flanders, Belgium

来源：

JOURNAL OF NEUROSCIENCE METHODS | 2016年 / 261卷

关键词：

Alpha-synuclein; High-content analysis; Aggregation; Cell death; Parkinson's disease; VECTOR-MEDIATED OVEREXPRESSION; HIGH-THROUGHPUT QUANTIFICATION; S18Y POLYMORPHISM; MUTATION ANALYSIS; HCA-VISION; GENE; NEURODEGENERATION; ASSOCIATION; CONSEQUENCES; PATHOGENESIS;

D O I：

10.1016/j.jneumeth.2015.11.009

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：117 / 127

页数：11

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