title: High-content analysis of α-synuclein aggregation and cell death in a cellular model of Parkinson's disease.
abstract: BACKGROUND
  Alpha-synuclein (α-SYN) aggregates represent a key feature of Parkinson's disease, but the exact relationship between α-SYN aggregation and neurodegeneration remains incompletely understood. Therefore, the availability of a cellular assay that allows medium-throughput analysis of α-SYN-linked pathology will be of great value for studying the aggregation process and for advancing α-SYN-based therapies.
  
  
  METHODS
  Here we describe a high-content neuronal cell assay that simultaneously measures oxidative stress-induced α-SYN aggregation and apoptosis.
  
  
  RESULTS
  We optimized an automated and reproducible assay to quantify both α-SYN aggregation and cell death in human SH-SY5Y neuroblastoma cells.
  
  
  METHODS
  Quantification of α-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 α-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 α-SYN aggregation and/or toxicity.
authors.affiliation: KU Leuven, Laboratory for Neurobiology and Gene Therapy, Kapucijnenvoer 33, Leuven B-3000, Flanders, Belgium.
authors.forename: Francesca
authors.initials: F
authors.lastname: Macchi
authors.affiliation: KU Leuven, Laboratory for Neurobiology and Gene Therapy, Kapucijnenvoer 33, Leuven B-3000, Flanders, Belgium.
authors.forename: Angélique
authors.initials: A
authors.lastname: Deleersnijder
authors.affiliation: KU Leuven, Laboratory for Neurobiology and Gene Therapy, Kapucijnenvoer 33, Leuven B-3000, Flanders, Belgium; Leuven Viral Vector Core, KU Leuven, Leuven B-3000, Flanders, Belgium.
authors.forename: Chris
authors.initials: C
authors.lastname: Van den Haute
authors.affiliation: KU Leuven, Department of Human Genetics, Flanders Interuniversity Institute of Biotechnology, Kapucijnenvoer 33, Leuven B-3000, Flanders, Belgium.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Munck
authors.affiliation: KU Leuven Campus Kulak Kortrijk, Public Health and Primary Care, Interdisciplinary Research Facility Life Sciences, Etienne Sabbelaan 53, Kortrijk B-8500, Flanders, Belgium.
authors.forename: Hans
authors.initials: H
authors.lastname: Pottel
authors.affiliation: KU Leuven, Laboratory for Neurobiology and Gene Therapy, Kapucijnenvoer 33, Leuven B-3000, Flanders, Belgium; Leuven Viral Vector Core, KU Leuven, Leuven B-3000, Flanders, Belgium.
authors.forename: Annelies
authors.initials: A
authors.lastname: Michiels
authors.affiliation: Laboratory for Molecular Virology and Gene Therapy, Department of Pharmaceutical and Pharmacological Sciences, KU Leuven, Leuven B-3000, Flanders, Belgium.
authors.forename: Zeger
authors.initials: Z
authors.lastname: Debyser
authors.affiliation: KU Leuven, Laboratory for Neurobiology and Gene Therapy, Kapucijnenvoer 33, Leuven B-3000, Flanders, Belgium; KU Leuven campus Kulak Kortrijk, Laboratory of Biochemistry, Interdisciplinary Research Facility Life Sciences, Etienne Sabbelaan 53, Kortrijk B-8500, Flanders, Belgium.
authors.forename: Melanie
authors.initials: M
authors.lastname: Gerard
authors.affiliation: KU Leuven, Laboratory for Neurobiology and Gene Therapy, Kapucijnenvoer 33, Leuven B-3000, Flanders, Belgium. Electronic address: veerle.baekelandt@med.kuleuven.be.
authors.forename: Veerle
authors.initials: V
authors.lastname: Baekelandt
mesh_terms: D017209:Apoptosis; D052160:Benzothiazoles; D015153:Blotting, Western; D002452:Cell Count; D018929:Cell Culture Techniques / Q000379:methods; D045744:Cell Line, Tumor; D002470:Cell Survival; D005822:Genetic Vectors; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D007150:Immunohistochemistry / Q000379:methods*; D007211:Indoles; D016086:Lentivirus / Q000235:genetics; D008856:Microscopy, Fluorescence / Q000379:methods; D018384:Oxidative Stress; D020734:Parkinsonian Disorders / Q000378:metabolism* / Q000473:pathology*; D066263:Protein Aggregation, Pathological; D055503:Protein Multimerization; D012984:Software; D013844:Thiazoles; D051844:alpha-Synuclein / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D052160:Benzothiazoles; D007211:Indoles; C497604:SNCA protein, human; D013844:Thiazoles; D051844:alpha-Synuclein; C009462:thioflavin T; C007293:DAPI
keywords: Aggregation; Alpha-synuclein; Cell death; High-content analysis; Parkinson's disease