The short-term effect of recombinant human leptin (rhleptin) on FSH and LH production (release + intracellular content) was studied in vitro using pituitary cells from male and female rainbow trout during the first gametogenesis cycle. In our rearing conditions, we found a direct action of rhleptin at the pituitary level, which depends on the sexual stage of the fish. No effect of rhleptin on FSH or LH release and cellular content could be detected in immature fish and post-ovulatory females. However, throughout the process of spermatogenesis and ovogenesis, high concentrations (0.5 and 1×10−6 M) of rhleptin stimulated FSH and LH release, without observable action on intracellular content of gonadotropins. A relatively constant response to rhleptin for FSH was observed throughout gonad maturation, while LH response tended to be higher at the first stages of gametogenesis (beginning of spermatogenesis and endogenous vitellogenesis). Preliminary results on the potential interaction of rhleptin and salmon GnRH (sGnRH) suggest a possible synergistic effect of high concentration of rhleptin (10−6 M) and sGnRH only at restricted phases of gonadal development when the gametogenetic process was already fully started (full spermatogenesis and early vitellogenesis). The direct action of leptin on FSH and LH release, evident only when gametogenesis had already started suggests that leptin is not the unique signal for the activation of the gonadotropic axis but requires a combined action with other promoting factors.