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This version published online on September 6, 2007
Endocrinology, doi:10.1210/en.2007-0961
A more recent version of this article appeared on December 1, 2007
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*Gene
*Compound via MeSH
*Substance via MeSH

Submitted on July 13, 2007
Accepted on August 30, 2007

KISSPEPTIN NEURONS IN THE ARCUATE NUCLEUS OF THE EWE EXPRESS BOTH DYNORPHIN A AND NEUROKININ B

Robert L. Goodman*, Michael N. Lehman, Jeremy T. Smith, Lique M. Coolen, Cleusa V.R. de Oliveira, Mohammad R. Jafarzadehshirazi, Alda Pereira, Javed Iqbal, Alain Caraty, Philippe Ciofi, and Iain J. Clarke

Department of Physiology and Pharmacology (R.L.G.), West Virginia University, Morgantown, West Virginia 26506-9229; Departments of Anatomy and Cell Biology (M.N.L.); Physiology and Pharmacology (L.M.C., C.V.R.D.), University of Western Ontario, London, Ontario, N6A 5C1, Canada; Department of Physiology (J.T.S., M.R.J., A.P., J.I., I.J.C.), Monash University, Victoria, Australia 3880; Animal Physiology Department (M.R.J.), Tehran University, Tehran, Iran; Unité Mixte de Recherche 6175 (A.C.), Institut National de la Recherche Agronomique/Centre National de la Recherche Scientifique, Université de Tours, Haras Nationaux, Institut Fédératif de Recherche 135, Nouzilly, France; and Inserm U862 (P.C.), Institut Francois Magendie, F-33077 Cedex, Bordeaux, France

* To whom correspondence should be addressed. E-mail: bgoodman{at}hsc.wvu.edu.

Kisspeptin is a potent stimulator of GnRH secretion that has been implicated in the feedback actions of ovarian steroids. In ewes, the majority of hypothalamic kisspeptin neurons are found in the arcuate nucleus (ARC), with a smaller population located in the preoptic area (POA). Most arcuate kisspeptin neurons express estrogen receptor-{alpha}, as do a set of arcuate neurons that contain both dynorphin and neurokinin B (NKB), suggesting that all three neuropeptides are co-localized in the same cells. In this study, we tested this hypothesis using dual-immunocytochemistry and also determined if kisspeptin neurons contain melanocyte-stimulating hormone (MSH) or agouti-related peptide (AGRP). To assess co-localization of kisspeptin and dynorphin, we used paraformaldehyde-fixed tissue from estrogen-treated ovariectomized ewes in the breeding season (n=5). Almost all ARC, but no POA, kisspeptin neurons contained dynorphin. Similarly, almost all ARC dynorphin neurons contained kisspeptin. In Exp. 2, we examined co-localization of kisspeptin and NKB in picric-acid fixed tissue collected from ovary-intact ewes (n=9). Over three-quarters of ARC kisspeptin neurons also expressed NKB, and a similar percentage of NKB neurons contained kisspeptin. In contrast, no kisspeptin neurons stained for MSH or AGRP. These data demonstrate that, in the ewe, a high percentage of ARC kisspeptin neurons also produce dynorphin and NKB, and we propose that a single subpopulation of ARC neurons contains all three neuropeptides. Because virtually all of these neurons express estrogen and progesterone receptors, they are likely to relay the feedback effects of these steroids to GnRH neurons to regulate reproductive function.
Key words: kisspeptin • dynorphin • neurokinin B • steroid feedback • GnRH




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