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Dopamine deficiency is mainly caused by apoptosis of dopaminergic nerve cells in the substantia nigra of themidbrain and the striatum and is an important pathologic basis of Parkinson’s disease (PD). Recent research has shownthat dynamin-related protein 1 (Drp1)-mediated aberrant mitochondrial fission plays a crucial role in dopaminergicnerve cell apoptosis. However, the upstream regulatory mechanism remains unclear. Our study showed that Drp1knockdown inhibited aberrant mitochondrial fission and apoptosis. Importantly, we found that ROCK1 was activated inan MPP+-induced PD cell model and that ROCK1 knockdown and the specific ROCK1 activation inhibitor Y-27632blocked Drp1-mediated aberrant mitochondrial fission and apoptosis of dopaminergic nerve cells by suppressing Drp1dephosphorylation/activation. Our in vivo study confirmed that Y-27632 significantly improved symptoms in a PDmouse model by inhibiting Drp1-mediated aberrant mitochondrial fission and apoptosis. Collectively, our findingssuggest an important molecular mechanism of PD pathogenesis involving ROCK1-regulated dopaminergic nerve cellapoptosis via the activation of Drp1-induced aberrant mitochondrial fission.