Protocol to investigate Parkinson's patient-derived dopaminergic neurons by live-cell microscopy and oxidized dopamine assays

Pingping Song; Dimitri Krainc

doi:10.1016/j.xpro.2024.102889

Protocol to investigate Parkinson's patient-derived dopaminergic neurons by live-cell microscopy and oxidized dopamine assays

Pingping Song, Dimitri Krainc^*

^*Corresponding author for this work

Neurology

Research output: Contribution to journal › Article › peer-review

Abstract

Dopaminergic neurons derived from human induced pluripotent stem cells recapitulate key pathogenic phenotypes observed in Parkinson's disease (PD). Here, we present a protocol to analyze oxidized dopamine and the recruitment of parkin onto synaptic vesicles in neurons derived from patients with mutations in parkin that cause autosomal recessive PD. We describe steps for neuronal differentiation, live-cell microscopy, detection of oxidized dopamine, and labeling of synaptic vesicles. These protocols can be applied to studies of other forms of genetic and sporadic forms of PD. For complete details on the use and execution of this protocol, please refer to Song et al.¹^,²

Original language	English (US)
Article number	102889
Journal	STAR Protocols
Volume	5
Issue number	1
DOIs	https://doi.org/10.1016/j.xpro.2024.102889
State	Published - Mar 15 2024

Funding

This work was supported by National Institutes of Health grants R01NS076054 , R37 NS096241 , and R35 NS122257 to D.K. Live-cell imaging work was performed by Wesley Peng at the Northwestern University Center for Advanced Microscopy generously supported by NCI CCSG P30 CA060553 awarded to the Robert H Lurie Comprehensive Cancer Center.

Keywords

Cell Biology
Neuroscience
Stem Cells

ASJC Scopus subject areas

General Neuroscience
General Biochemistry, Genetics and Molecular Biology
General Immunology and Microbiology

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10.1016/j.xpro.2024.102889

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abstract = "Dopaminergic neurons derived from human induced pluripotent stem cells recapitulate key pathogenic phenotypes observed in Parkinson's disease (PD). Here, we present a protocol to analyze oxidized dopamine and the recruitment of parkin onto synaptic vesicles in neurons derived from patients with mutations in parkin that cause autosomal recessive PD. We describe steps for neuronal differentiation, live-cell microscopy, detection of oxidized dopamine, and labeling of synaptic vesicles. These protocols can be applied to studies of other forms of genetic and sporadic forms of PD. For complete details on the use and execution of this protocol, please refer to Song et al.1,2",

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AB - Dopaminergic neurons derived from human induced pluripotent stem cells recapitulate key pathogenic phenotypes observed in Parkinson's disease (PD). Here, we present a protocol to analyze oxidized dopamine and the recruitment of parkin onto synaptic vesicles in neurons derived from patients with mutations in parkin that cause autosomal recessive PD. We describe steps for neuronal differentiation, live-cell microscopy, detection of oxidized dopamine, and labeling of synaptic vesicles. These protocols can be applied to studies of other forms of genetic and sporadic forms of PD. For complete details on the use and execution of this protocol, please refer to Song et al.1,2

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Protocol to investigate Parkinson's patient-derived dopaminergic neurons by live-cell microscopy and oxidized dopamine assays

Abstract

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Keywords

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