The effect of vitamin D deficiency on the fertility and reproductive capacity of male rats was investigated. Male weanling rats were fed vitamin D-deficient or vitamin D-replete diets until maturity, and mated to age-matched, vitamin D-replete females. Vitamin D-deficient males were capable of reproduction. However, successful matings, i.e., presence of sperm in the vaginal tract of the female, by vitamin D-deficient males were reduced by 45% when compared to matings by vitamin D-replete males. Fertility (successful pregnancies in sperm-positive females) was reduced by 73% in litters from vitamin D-deficient male inseminations when compared to litters from females inseminated by vitamin D-replete males. These results demonstrate that vitamin D and its metabolites are necessary for normal reproductive functions in the male rat.