Fluorine, an active element which is widely distributed in the environment, has been demonstrated to have adverse effects on male reproductive systems at a high dose. The objective of the present study was to explore the key role of cysteine-rich secretory protein (CRISP)2 in fluoride-induced male reproductive toxicity by investigating, in rats, the effect of the administration of fluoride on the expression of CRISP2 in the testes, in the immature sperm in the initial segment of the epididymis, and in the mature sperm in the tail of the epididymis. In this study, after mating, female pregnant rats were randomly divided into four groups: control (distilled water) and NaF (25, 50, and 100 mg L-1) groups. After weaning, the dosage was continued for 8 weeks for 36 puppies. After exposure, the histological structure of the testis was assessed using HE, and the localization and protein expression of CRISP2 in the rat testis and epididymal sperm were evaluated by indirect immunofluorescence. In addition, sperm survival and mitochondrial transmembrane potential were evaluated. The results showed that fluoride impaired the histological structure of the testis, decreased the level of CRISP2, and reduced sperm survival and mitochondrial transmembrane potential. Together, our present data suggest that the decreased fertility in rats affected by fluorosis might be, in part, through the low expression of CRISP2 in testis and epididymal sperm.