Nitric oxide signaling in ghrelin-induced LH release from goldfish pituitary cells

Among its many known functions, ghrelin has been proposed to participate in the regulation of reproduction; however, its effect on pituitary LH release is controversial, especially in mammals. In the goldfish, ghrelin directly stimulates pituitary LH release via increased entry of calcium through voltage sensitive channels and activation of protein kinase C. Nitric oxide (NO) is an important signaling molecule in many physiological systems including hormone regulation at the level of the pituitary. Goldfish pituitary cells and extracts have previously been reported to express immunoreactivity for inducible and neuronal NO synthase (iNOS and nNOS). In this study, we determined if NO is involved in goldfish ghrelin (gGRLN19)-induced LH release from primary cultures of dispersed goldfish pituitary cells in column perifusion. Treatment with the NO scavenger PTIO significantly decreased gGRLN19-induced LH release and co-treatment with the NO donor SNP and gGRLN19 did not induce an additive increase in LH release, suggesting that NO is critical to gGRLN19 stimulation of LH release in goldfish pituitary cells. Further work examined the involvement of the NOS using the NOS isoform-selective inhibitors 1400 W, 7-Ni, and AGH. While 1400 W (selective for iNOS) and AGH (selective for iNOS and nNOS) abolished gGRLN19-induced LH release from goldfish pituitary cells, 7-Ni (selective for nNOS and endothelial NOS) had no significant effect on this stimulation. Our results indicate, for the first time in a teleost species, that gGRLN19-induced LH release from pituitary cells is NO-dependent and likely involves iNOS, adding to the understanding of GRLN intracellular signaling in general and specifically to the regulation of LH release from the pituitary.

► 1st demonstration of NOS/NO involvement in ghrelin-induced LH release in any teleost. ► Identification of iNOS as the most likely NOS isoform involved. ► Implication for cell-type specific NOS involvement in ghrelin pituitary signalling.