Protocol for generation of PD modeling induced neurons and detection of α-synuclein forms
Year: 2024
Type: article
Abstract: Alpha-synuclein (α-Syn) is an important molecule in the pathogenesis of Parkinson’s disease and Alzheimer’s disease-related dementias such as Lewy body dementia, forming multiple pathological species. In vitro disease models, including human neurons and α-Syn-transfected cells, are instrumental to understand synucleinopathies or test new therapies. Here, we provide a detailed protocol to generate human neurons derived from induced pluripotent stem cells (iPSCs), and HEK cells, with α-Syn mutations. We also describe multiple assays to determine the various α-Syn forms. • Methods to generate PD cell models with multiple α-Syn species in vitro • Protein extraction and media collection from induced neurons and transfected cells • Guide for analysis of intracellular and extracellular levels of α-Syn via ELISA • Instructions for investigating major α-Syn species using immunoblotting Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Alpha-synuclein (α-Syn) is an important molecule in the pathogenesis of Parkinson’s disease and Alzheimer’s disease-related dementias such as Lewy body dementia, forming multiple pathological species. In vitro disease models, including human neurons and α-Syn-transfected cells, are instrumental to understand synucleinopathies or test new therapies. Here, we provide a detailed protocol to generate human neurons derived from induced pluripotent stem cells (iPSCs), and HEK cells, with α-Syn mutations. We also describe multiple assays to determine the various α-Syn forms.
Source: STAR Protocols
Authors: Francesco V. Nardi, Gulimiheranmu Maisumu, You Zhou, Bo Liang, Abraam M. Yakoub
Language: English
Subfield: Neurology
Field: Medicine
Domain: Health Sciences
Open Access status: gold
Estimated APC paid: $1,500
Funders: Foundation for the National Institutes of Health, Massachusetts Institute of Technology, Northwestern University, National Institutes of Health, Whitehead Institute for Biomedical Research, Massachusetts Institute of Technology, National Institute on Aging, National Heart, Lung, and Blood Institute
OpenAlex ID: W4404397409