In the present study, using Northern blot analysis, we demonstrated that the level of iNOS mRNA in the ovary of immature rats decreased after 6 h of pregnant mare serum gonadotropin administration and recovered gradually up to the untreated level 48 h after the administration. Both in situ hybridization and immunohistochemistry revealed that iNOS mRNA and protein was predominantly localized in granulosa cells in most of immature follicles, but not in mature follicles with an antrum, which was a consistent finding regardless of gonadotropin treatment. Furthermore, we found that cultured granulosa cells had the ability to express iNOS mRNA in the presence of cytokines such as tumor necrosis factor-alpha, interleukin-1 beta and interferon-gamma, which are inherently detectable in the ovary. These results raise the possibility that locally produced NO synthesized by iNOS may be involved in the developmental status of ovarian follicles in concert with gonadotropin.