Prenatal androgen blockade with the antiandrogen flutamide inhibits the inguinoscrotal phase of testicular descent. The evidence suggests that androgens may act indirectly via the sexually dimorphic genitofemoral nerve (GFN) to control this phase. Rats were exposed to flutamide on gestational days 16 through 19. Seven-day-old rats were subjected to retrograde fluorescent labelling of the GFN combined with immunohistochemistry for calcitonin gene-related peptide (CGRP), a neurotransmitter found in the GFN. Fluorescent-labelled and CGRP-immunoreactive neurons in the GFN spinal nucleus were quantified. Sexual dimorphism of the GFN nucleus was absent in the flutamide-treated rats but obviously present in control rats. Furthermore, control male nuclei had 24% more CGRP-immunoreactive neurons and 12% more fluorescent-labelled neurons than did flutamide-treated male nuclei. This study shows that prenatal androgen blockade with flutamide inhibits masculinization of the GFN, with significant reduction of its CGRP content. This supports the proposal that androgens act via the GFN, with CGRP as the second messenger, to control inguinoscrotal testicular descent.