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

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.