Recently, the structures of two forms of inhibin present in human follicular fluid were elucidated from the corresponding cDNA sequences. Using specific antisera generated against the alpha-chain common to both forms, we have examined the cellular localization of inhibin in the male and female rat gonads and in human placental tissue. Specific alpha-inhibin immunoreactivity was localized within the Sertoli cells of a number of tubules in each testes section. However, other adjacent tubules were unstained suggesting a stage-specific production of inhibin. Intense immunostaining was observed in the granulosa cells of ovarian follicles at various stages but not in the thecal cells. Immunostaining was present in the human placenta and limited to the cytotrophoblast cells, suggesting a role of inhibin during pregnancy. The present study demonstrates the probable site of production of inhibin in the gonads and placenta and further implicates this important factor as a key regulator of reproductive functions.