Mouse spermatozoa are able to produce ATP and sustain vigorous motility via two metabolic pathways, glycolysis and oxidative phosphorylation (OXPHOS). Numerous studies showed that glycolysis is the main source of ATP required to sustain the motility of mouse sperm and that an active glycolytic pathway is essential for sperm motility in this species. All available evidence has been gathered from strains of the laboratory mouse (Mus musculus). However, comparative studies between several mouse species (genus Mus) have revealed that (a) there is considerable interspecific variability regarding sperm motility and swimming velocity within this group, (b) these variations are strongly associated to divergences in sperm ATP content, and (c) M. musculus is at the lower end of the distribution of these sperm parameters. Since M. musculus sperm metabolism relies heavily on glycolysis, a less efficient ATP generator than OXPHOS, we asked whether variations in ATP production and sperm swimming velocity between Mus species could relate to species differences in these two metabolic pathways. In this study we compared the relative reliance on the usage of glycolysis and OXPHOS as ATP source to sustain sperm motility between Mus species that have significantly different basal sperm performance parameters. We found significant interspecific variations in the glycolysis and OXPHOS rates between different Mus species. The species with higher OXPHOS/glycolysis ratio showed higher sperm ATP content, motility index, and swimming velocity. Moreover, inhibition of OXPHOS significantly reduced ATP content and performance parameters in the species with a highly respiratory sperm, while having no effect in the species with highly glycolytic sperm. Overall, our findings support the idea that the diversity in sperm performance within the genus Mus can be explained by variations in the relative usage ratio of these two metabolic pathways.