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The Secretogranin II-Derived Peptide Secretoneurin Stimulates Luteinizing Hormone Secretion from Gonadotrophs

Department of Biology (E.Z., V.L.T.), Centre for Advanced Research in Environmental Genomics, and the Chronic Disease Program (A.B., A.C.), Ottawa Health Research Institute, Ottawa, Ontario, Canada K1Y 4E9; School of Biological Sciences (A.O.L.W., W.K.), The University of Hong Kong, Hong Kong SAR, China; Laboratorios de Ictiofisiologia y Acuicultura (G.C.L., G.M.S.) and Neuroendocrinología Comparada (L.F.C.), Instituto de Investigaciones Biotecnológicas-Instituto Tecnologico Chascomús, Provincia de Buenos Aires, C1428DN Buenos Aires, Argentina; and Department of Biological Sciences (C.L.G., J.P.C.), University of Alberta, Edmonton, Alberta, Canada T6G 2E9

Address all correspondence and requests for reprints to: Vance L. Trudeau, Department of Biology, Centre for Advanced Research in Environmental Genomics, Ottawa Health Research Institute, Ottawa, Ontario, Canada K1Y 4E9. E-mail: trudeauv{at}uottawa.ca.

Secretoneurin (SN) is a 33- to 34-amino acid neuropeptide derived from secretogranin-II, a member of the chromogranin family. We previously synthesized a putative goldfish (gf) SN and demonstrated its ability to stimulate LH release in vivo. However, it was not known whether goldfish actually produced the free SN peptide or whether SN directly stimulates LH release from isolated pituitary cells. Using a combination of reverse-phase HPLC and mass spectrometry analysis, we isolated for the first time a 34-amino acid free gfSN peptide from the whole brain. Moreover, Western blot analysis indicated the existence of this peptide in goldfish pituitary. Immunocytochemical localization studies revealed the presence of SN immunoreactivity in prolactin cells of rostral pars distalis of the anterior pituitary. Additionally, we found that magnocellular cells of the goldfish preoptic region are highly immunoreactive for SN. These neurons send heavily labeled projections that pass through the pituitary stalk and innervate the neurointermediate and anterior lobes. In static 12-h incubation of dispersed pituitary cells, application of SN antiserum reduced LH levels, whereas 1 and 10 nM gfSN, respectively, induced 2.5-fold (P < 0.001) and 1.9-fold (P < 0.01) increments of LH release into the medium, increases similar to those elicited by 100 nM concentrations of GnRH. Like GnRH, gfSN elevated intracellular Ca²⁺ in identified gonadotrophs. Whereas we do not yet know the relative contribution of neural SN or pituitary SN to LH release, we propose that SN could act as a neuroendocrine and/or paracrine factor to regulate LH release from the anterior pituitary.