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Cloning and Localization of Three Forms of Gonadotropin-Releasing Hormone, Including the Novel Whitefish Form, in a Salmonid, Coregonus clupeaformis¹

Department of Biology,³ University of Victoria, Victoria, British Columbia, Canada V8W 3N5 Department of Zoology,⁴ University of Manitoba, Winnipeg, Manitoba, Canada R3T 2N2 Freshwater Institute,⁵ Fisheries and Oceans Canada, Winnipeg, Manitoba, Canada R3T 2N6

Cells containing different GnRH peptides currently are thought to have distinct locations and functions in the brain. Lake whitefish is the first salmonid species to have three forms of GnRH peptide in contrast to later-evolving salmonids (salmon and trout) in which only two forms have been identified. Our objective was to isolate the cDNAs that code for these transcripts and to localize the transcripts for the three forms of GnRH in adult lake whitefish brain. Also, we provide phylogenetic analysis of these three whitefish genes based on their preprohormone sequence. From whitefish we isolated cDNAs encoding chicken (c)GnRH-II, salmon (s)GnRH, and the novel whitefish (wf)GnRH. The three cDNAs each encode only one GnRH and are placed in separate groups with phylogenetic analysis. A combination of in situ hybridization and immunocytochemistry with two antisera revealed neurons that expressed protein and/or mRNA for cGnRH-II in the midbrain and hindbrain; sGnRH in the olfactory nerve and bulb, ventral telencephalon, and preoptic area; and wfGnRH in the same latter two brain regions and the hypothalamus. Thus, in the anterior brain, cells containing sGnRH and wfGnRH were in the same brain areas but not at identical locations in the ventral telencephalon and preoptic area. Based on our results, we speculate that both sGnRH and wfGnRH have gonadotropin-releasing roles in the lake whitefish brain.

¹ Supported by the Canadian Institutes of Health Research grant (N.M.S.) and fellowship (B.A.A.); Natural Sciences and Engineering Research Council of Canada grant OGP0007576 (T.J.H.); Fonds pour la formation de Chercheurs et l'Aide à la Recherche (Québec); and the University of Manitoba Graduate Fellowship (F.L.).

² Correspondence: Nancy M. Sherwood, Department of Biology, 3800 Finnerty Road, University of Victoria, Victoria, BC, Canada, V8W 3N5. FAX: 250 721 7120; nsherwoo{at}uvic.ca

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