Disruption of Odf4 led to male infertility by impairing mitochondrial function and fertilization capacity. Mechanistically, Lrrc71 interacted with fertilization-related proteins, including ADAM2, CALR, and ZP3R. Furthermore, Lrrc71 deficiency resulted in reduced levels of mitochondrial function-associated proteins. Collectively, our findings establish Odf4 as a central regulator of mitochondrial function and fertilization, providing mechanistic insights into the pathogenesis of infertility caused by human Odf4 mutations.