Animal development requires precise regulation of mitochondrial fusion and fission to ensure optimal mitochondrial function and cellular activity. Perturbations in the balance between these processes can lead to mitochondrial fragmentation with loss of normal membrane potential in individual mitochondria (ΔΨm). In this study, we showed that MIRO-1 is stochastically elevated in individual fragmented mitochondria, potentially acting as a compensatory mechanism to maintain ΔΨm. This observation is supported by our finding of higher levels of ΔΨm in fragmented mitochondria in fzo-1 mutants and animals after wounding. Moreover, MIRO-1 interacts with the voltage-dependent anion channel (VDAC-1), a crucial component of the mitochondrial ion channel located on the outer mitochondrial membrane. We find that the interaction between MIRO-1 and VDAC-1 depends on the residue E473 in MIRO-1 and K163 in VDAC-1. Disrupting their interaction by E473G point mutation reduces the ΔΨm. Our findings suggest that MIRO-1 interacts with VDAC-1 to regulate ΔΨm to maintain mitochondrial activity and animal health through regulating VDAC-1 activity. This study provides insight into the mechanisms underlying the stochastic maintenance of residual ΔΨm in fragmented mitochondria.