Parkinson’s disease (PD) is a debilitating neurodegenerative disorder characterized by the preferential loss of dopaminergic neurons in the Substantia nigra resulting in a dopamine (DA) depletion in the striatum and subsequential motor impairment in patients. Increasing evidence has linked mitochondrial dysfunction to PD; however, the underlying molecular mechanisms remain unclear. Recently, mutations in CHCHD2 gene have been identified in PD patients. CHCHD2 belongs to a family of evolutionary conserved mitochondrial protein, which act as regulators of mitochondrial ultrastructure and dynamics. Initial studies reported that CHCHD2 is localized in mitochondrial intermembrane space, where physically interacts with the family member, CHCHD10. Importantly, CHCHD10 mutations have been implicated in amyotrophic lateral sclerosis and frontotemporal dementia. To date, the role of CHCHD2 and CHCHD10 in mitochondria is still unknown. To investigate CHCHD2 function in vivo, we here employed a whole-body CHCHD2 knockout mouse and performed a variety of behavioral, molecular, and biochemical analyses. We found that the CHCHD2 loss causes mild motor deficits in aged mice and decreased DA levels in the striatum. In addition, we showed that CHCHD2 ablation leads to discernible effect on oxidative phosphorylation. In mouse tissues, CHCHD2 and CHCHD10 do not exist as monomers but form a high molecular weight complex, which accumulates in response to severe mitochondrial damage both in vivo and in vitro. Collectively, our results underscore the critical role of CHCHD2-CHCHD10 protein complex in maintaining mitochondrial function, offering novel insights in understanding the contribution of mitochondria to neuronal survival.