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Merck
  • Optogenetic induction of alpha-synuclein aggregation in human dopaminergic neurons to model Parkinson's disease pathology.

Optogenetic induction of alpha-synuclein aggregation in human dopaminergic neurons to model Parkinson's disease pathology.

STAR protocols (2023-09-24)
Eun A Ra, Min Seong Kim, Gabsang Lee
摘要

Alpha-synuclein (α-syn) aggregation is a principal factor in Parkinson's disease (PD) onset. Here, we present a protocol for optogenetic induction of α-syn aggregation in human midbrain dopaminergic (mDA) neurons, facilitating a detailed PD pathology study. We describe steps for nucleofection of the opto-α-syn construct, single colony selection and validation, alongside mDA neuron differentiation and rapid induction of toxic α-syn aggregates via blue light. This establishes a potent human induced pluripotent-stem-cell-based platform for PD drug testing and validation. For complete details on the use and execution of this protocol, please refer to Kim et al. (2023).1.

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Sigma-Aldrich
嘌呤霉素 二盐酸盐 来源于白色链球菌, powder, BioReagent, suitable for cell culture
Sigma-Aldrich
N6,2′-O-二丁酰基腺苷 3′,5′-环单磷酸 钠盐, ≥96% (HPLC), powder
Sigma-Aldrich
抗-聚集α-突触核蛋白抗体,克隆5G4, clone 5G4, from mouse, purified by affinity chromatography
Sigma-Aldrich
抗-mCherry, from rabbit