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Image-based quantitation of kainic acid-induced excitotoxicity as a model of neurodegeneration in Human iPSC-Derived Neurons

Citation

Talbot, J and Chear, S and Phipps, A and Pebay, A and Hewitt, AW and Vickers, JC and King, AE and Cook, AL, Image-based quantitation of kainic acid-induced excitotoxicity as a model of neurodegeneration in Human iPSC-Derived Neurons, Methods in Molecular Biology ISSN 1064-3745 (2021) [Refereed Article]


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DOI: doi:10.1007/7651_2021_421

Abstract

Excitotoxicity is a feature of many neurodegenerative diseases and acquired forms of neural injury that is characterized by disruption of neuronal morphology. This is typically seen as beading and fragmentation of neurites when exposed to excitotoxins such as the AMPA receptor agonist kainic acid, with the extent to which these occur used to quantitate neurodegeneration. Induced pluripotent stem cells (iPSCs) provide a means to generate human neurons in vitro for mechanistic studies and can thereby be used to investigate how cells respond to excitotoxicity and to identify or test potential neuroprotective agents. To facilitate such studies, we have optimized a protocol for human iPSC differentiation to mature neurons in a 96-well plate format that enables image-based quantitation of changes to neuron morphology when exposed to kainic acid. Our protocol assays neuron morphology across seven excitotoxin concentrations with multiple control conditions and is ideally suited to comparison of neurons generated through differentiation of two isogenic iPSC lines in a single plate. We have included detailed step-by-step protocols for neural stem cell differentiation, neuronal maturation and exposure to kainic acid treatment, as well as different approaches to image-based quantitation that involve immunofluorescence or phase-contrast microscopy.

Item Details

Item Type:Refereed Article
Keywords:stem cell, neurodegeneration, disease model, image analysis
Research Division:Biological Sciences
Research Group:Biochemistry and cell biology
Research Field:Biochemistry and cell biology not elsewhere classified
Objective Division:Health
Objective Group:Clinical health
Objective Field:Prevention of human diseases and conditions
UTAS Author:Talbot, J (Dr Jana Talbot)
UTAS Author:Chear, S (Ms Sueanne Chear)
UTAS Author:Phipps, A (Dr Andrew Phipps)
UTAS Author:Hewitt, AW (Professor Alex Hewitt)
UTAS Author:Vickers, JC (Professor James Vickers)
UTAS Author:King, AE (Professor Anna King)
UTAS Author:Cook, AL (Associate Professor Tony Cook)
ID Code:146574
Year Published:2021
Deposited By:Wicking Dementia Research and Education Centre
Deposited On:2021-09-14
Last Modified:2021-09-14
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