The present study was undertaken to evaluate the effects of the chemosterilant 2,4-dinitrotoluene (DNT) on the rat testis. Adult male rats were fed control, or 0.1%, or 0.2% DNT for 3 weeks. An ultrastructural study of the testes was performed, serum was assayed for testosterone and gonadotropins, and sperm reserve count was determined. A marked change in Sertoli cell morphology was found after 3 weeks of 0.2% DNT exposure. Varying sized vesicles associated with swollen mitochondria and distended endoplasmic reticulum were visible in cells from DNT-treated animals. Circulating levels of follicle stimulating hormone and luteinizing hormone were increased in 0.2% DNT-treated animals. Reduced weights of the epididymides and decreased epididymal sperm reserves were observed in DNT-treated animals. These results indicate that DNT is capable of inducing testicular injury, of directly or indirectly disturbing pituitary function, and of exerting a toxic effect at the late stages of spermatogenesis. These findings suggest that a locus of DNT action is the Sertoli cell, resulting in both inhibition of spermatogenesis and changes in testicular-pituitary endocrine activity.