Caspase-2 and caspase-8 trigger caspase-3 activation following 6-OHDA-induced stress in human dopaminergic neurons differentiated from ReNVM stem cells

AbstractObjectives: The purpose of the study was to establish a suitable model to study Parkinson’s disease (PD) pathogenesis in differentiated dopaminergic neurons (dDCN). The specific aim was to demonstrate the involvement of the caspase family and to identify specific caspases which are activated by 6-hydroxydopamine (6OHD) treatment leading to death of dDCN.Methods: ReNcell VM cells were differentiated into dDCN and were exposed to 6OHD to induce stress. Western blot (WB) and double immunofluorescent analyses of caspases-2, -3, and -8 were carried out in untreated and 6OHD-treated dDCN. zVADfmk was used to determine if it could inhibit activation of caspases-2, -3, and -8 in dDCN following 6OHD-mediated stress.Results: Our immunofluorescent and WB data showed that 6OHD triggered caspases-2 and -8 activation which in turn activated caspase-3 leading to death of dDCN. Additionally, WB analysis revealed that caspases-2, -3, and -8 activation was reduced by zVADfmk in 6OHD-treated cells.Discussion: The st...