Leah, T., Vazquez-Villaseñor, I., Ferraiuolo, L. et al. (2 more authors) (2021) A Parkinson’s disease-relevant mitochondrial and neuronal morphology high-throughput screening assay in LUHMES cells. Bio-protocol, 11 (1). ISSN 2331-8325
Abstract
Parkinson’s disease is a devastating neurodegenerative disorder affecting 2-3% of the population over 65 years of age. There is currently no disease-modifying treatment. One of the predominant pathological features of Parkinson’s disease is mitochondrial dysfunction, and much work has aimed to identify therapeutic compounds which can restore the disrupted mitochondrial physiology. However, modelling mitochondrial dysfunction in a disease-relevant model, suitable for screening large compound libraries for ameliorative effects, represents a considerable challenge. Primary patient derived cells, SHSY-5Y cells and in vivo models of Parkinson’s disease have been utilized extensively to study the contribution of mitochondrial dysfunction in Parkinson’s. Indeed many studies have utilized LUHMES cells to study Parkinson’s disease, however LUHMES cells have not been used as a compound screening model for PD-associated mitochondrial dysfunction previously, despite possessing several advantages compared to other frequently used models, such as rapid differentiation and high uniformity (e.g., in contrast to iPSC-derived neurons), and relevant physiology as human mesencephalic tissue capable of differentiating into dopaminergic-like neurons that highly express characteristic markers. After previously generating GFP+-LUHMES cells to model metabolic dysfunction, we report this protocol using GFP+-LUHMES cells for high-throughput compound screening in a restoration model of PD-associated mitochondrial dysfunction. This protocol describes the use of a robust and reproducible toxin-induced GFP+-LUHMES cell model for high throughput compound screening by assessing a range of mitochondrial and neuronal morphological parameters. We also provide detailed instructions for data and statistical analysis, including example calculations of Z’-score to assess statistical effect size across independent experiments.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2021 The Authors. |
Keywords: | Parkinson’s disease; Mitochondria; Compound screening; LUHMES |
Dates: |
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Institution: | The University of Sheffield |
Academic Units: | The University of Sheffield > Sheffield Teaching Hospitals |
Depositing User: | Symplectic Sheffield |
Date Deposited: | 15 Jan 2021 12:06 |
Last Modified: | 18 Jan 2021 07:11 |
Status: | Published |
Publisher: | Bio-Protocol LLC |
Refereed: | Yes |
Identification Number: | 10.21769/bioprotoc.3881 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:170086 |