PT  - JOURNAL ARTICLE
AU  - Peter A. Barbuti
AU  - Bruno FR. Santos
AU  - Paul M. Antony
AU  - Francois Massart
AU  - Gérald Cruciani
AU  - Claire M. Dording
AU  - Lukas Pavelka
AU  - Yong-Jun Kwon
AU  - Rejko Krüger
TI  - Gene-corrected Parkinson’s disease neurons show the A30P alpha-synuclein point mutation leads to reduced neuronal branching and function
AID  - 10.1101/2020.11.05.369389
DP  - 2020 Jan 01
TA  - bioRxiv
PG  - 2020.11.05.369389
4099  - http://biorxiv.org/content/early/2020/11/06/2020.11.05.369389.short
4100  - http://biorxiv.org/content/early/2020/11/06/2020.11.05.369389.full
AB  - Parkinson’s disease is characterised by the degeneration of A9 dopaminergic neurons and the pathological accumulation of alpha-synuclein. In a patient-derived stem cell model, we have generated dopaminergic neurons from an individual harbouring the p.A30P SNCA mutation and compared those neurons against gene-corrected isogenic control cell lines. We have used confocal microscopy to assess the neuronal network, specifically segmenting dopaminergic neurons and have identified image-based phenotypes showing axonal impairment and reduced neurite branching. We show using multi-electrode array (MEA) technology that the neurons carrying the endogenous p.A30P alpha-synuclein mutation are functionally impaired and identified mitochondrial dysfunction as a pathogenic cellular phenotype. We report that against gene-corrected isogenic control cell lines the neurons carrying the p.A30P SNCA mutation have a deficit and are susceptible to the mitochondrial toxin and environmental pesticide Rotenone. Our data supports the use of isogenic cell lines in identifying image-based pathological phenotypes that can serve as an entry point for future disease modifying compound screenings and drug discovery strategies.Competing Interest StatementThe authors have declared no competing interest.