Participation of arachidonic acid metabolism in gonadotropin-releasing hormone stimulation of goldfish gonadotropin release

Two intraperitoneal injections of a mammalian gonadotropin-releasing hormone (GnRH) analog, [D-Ala6, Pro9-N-ethylamide]-GnRH (mGnRHa; 0.1 micrograms/g), at 12-hr intervals increased serum gonadotropin (GTH) levels in sexually mature and sexually regressed female goldfish 2 and 6 hr after the second injection. This serum GTH response was decreased by the coinjection of a lipoxygenase enzyme inhibitor, nordihydroguaiaretic acid (NDGA: 0.1 to 10 micrograms/g) at the time of the second mGnRHa application. In static cultures of dispersed goldfish pituitary cells, 1-100 microM arachidonic acid (AA) and 0.1-1000 nM [Trp7, Leu8]-GnRH (salmon GnRH, sGnRH) and [D-Arg6, Pro9-N-ethylamide]-sGnRH (sGnRHa) caused dose-dependent increases in GTH release. Additions of 1-40 microM NDGA reduced the sGnRH-stimulated GTH release in a dose-dependent manner, and completely inhibited the GTH response to increasing concentrations of AA. NDGA 40 microM also decreased the elevated GTH levels induced by sGnRHa treatment. Exposure to 10 microM 5,8,11,14-eicosatetraynoic acid, an inhibitor with mixed action on lipoxygenase and cyclooxygenase enzymes, reduced the dose-dependent GTH response to sGnRH and AA. In contrast, coincubation with another cyclooxygenase blocker, indomethacin, at 10 microM, did not alter AA and sGnRH-induced GTH release. These results provide in vivo and in vitro evidence for the participation of AA metabolism in mediating GnRH-stimulated GTH release in the goldfish. The importance of AA metabolism through the lipoxygenase pathway is also indicated.