Hypogonadotropic hypogonadism in mice lacking a functional <em>Kiss1</em> gene

Xavier de Tassigny

Lisa Fagg

John Dixon

Kate Day

Harry Leitch

Alan Hendrick

Dirk Zahn

Isabelle Franceschini

Alain Caraty+3 authors

*Reproductive Physiology Group, Department of Physiology, Development, and Neuroscience, University of Cambridge, Cambridge CB2 3EG, United Kingdom;

^{Paradigm Therapeutics Ltd. (now Takeda Cambridge Ltd.), 418 Cambridge Science Park, Milton Road, Cambridge CB4 0PA, United Kingdom;}

^{Unité de Physiologie de la Reproduction et des Comportements, Unité Mixte de Recherche 6175, Institut National de la Recherche Agronomique/Centre National de la Recherche Scientifique/Université Tours, 37380 Nouzilly, France; and}

^{BC Cancer Agency, 675 West Tenth Avenue, Vancouver, BC, Canada V5Z 1LR}

^{To whom correspondence should be addressed. E-mail:}ku.ca.mac@32chw

Communicated by Etienne-Emile Baulieu, Collège de France, Le Kremlin-Bicetre, France, May 5, 2007.

Author contributions: X.d.A.d.T. and W.H.C. designed research; X.d.A.d.T., L.A.F., J.P.C.D., K.D., H.G.L., A.G.H., I.F., D.Z., and W.H.C. performed research; I.F., A.C., and M.B.L.C. contributed new reagents/analytic tools; X.d.A.d.T., J.P.C.D., I.F., A.C., S.A.J.R.A., and W.H.C. analyzed data; and X.d.A.d.T., L.A.F., and W.H.C. wrote the paper.

Received 2006 Dec 12

Abstract

The G protein-coupled receptor GPR54 (AXOR12, OT7T175) is central to acquisition of reproductive competency in mammals. Peptide ligands (kisspeptins) for this receptor are encoded by the Kiss1 gene, and administration of exogenous kisspeptins stimulates hypothalamic gonadotropin-releasing hormone (GnRH) release in several species, including humans. To establish that kisspeptins are the authentic agonists of GPR54 in vivo and to determine whether these ligands have additional physiological functions we have generated mice with a targeted disruption of the Kiss1 gene. Kiss1-null mice are viable and healthy with no apparent abnormalities but fail to undergo sexual maturation. Mutant female mice do not progress through the estrous cycle, have thread-like uteri and small ovaries, and do not produce mature Graffian follicles. Mutant males have small testes, and spermatogenesis arrests mainly at the early haploid spermatid stage. Both sexes have low circulating gonadotropin (luteinizing hormone and follicle-stimulating hormone) and sex steroid (β-estradiol or testosterone) hormone levels. Migration of GnRH neurons into the hypothalamus appears normal with appropriate axonal connections to the median eminence and total GnRH content. The hypothalamic–pituitary axis is functional in these mice as shown by robust luteinizing hormone secretion after peripheral administration of kisspeptin. The virtually identical phenotype of Gpr54- and Kiss1-null mice provides direct proof that kisspeptins are the true physiological ligand for the GPR54 receptor in vivo. Kiss1 also does not seem to play a vital role in any other physiological processes other than activation of the hypothalamic–pituitary–gonadal axis, and loss of Kiss1 cannot be overcome by compensatory mechanisms.

Keywords: Gpr54, kisspeptin, mouse, puberty

Abstract

Neuroendocrine events within the hypothalamus control sexual maturation and seasonal breeding in mammals (1). The gonadotropin-releasing hormone (GnRH)-induced secretion of the gonadotropic hormones luteinizing hormone (LH) and follicle-stimulating hormone (FSH) from the anterior pituitary is essential to invoke puberty and maintain reproductive function. The G protein-coupled receptor GPR54 (2) is a key protein involved in the pubertal activation of the hypothalamic pituitary gonadal axis because mice and humans with mutations in this receptor are sterile with hypogonadotropic hypogonadism (3 –7).

A series of overlapping peptide ligands (kisspeptins) for the GPR54 receptor are produced by the Kiss1 gene (8 –10). Kiss1 mRNA is expressed in hypothalamic regions that regulate gonadotropin secretion including the anteroventral periventricular nucleus (AVPV), the periventricular nucleus, and the arcuate nucleus (ARC) (11). Kiss1 expression increases at puberty in rodents (12 –14) and primates (15) and fluctuates during the rat estrous cycle (12, 16). Kiss1 expression is also subject to differential regulation by sex steroids, providing a plausible mechanism for the feedback control of gonadotropin secretion by these hormones (17, 18). Administration of kisspeptins potently stimulates gonadotropin secretion in several mammalian species (11, 12, 19 –24). These effects are likely mediated by a direct action on GnRH neurons, which express the GPR54 receptor (13, 21, 25). Kisspeptins can stimulate GnRH release from explanted rat hypothalamic fragments (20, 26) and after central injection in sheep (25). Kisspeptin immunoreactive neurons have been shown to project fibers directly onto GnRH neurons (14, 24). Finally, direct electrophysiological studies on GnRH-GFP neurons in mice revealed that kisspeptin stimulates a robust and long-lasting depolarization in most of the GnRH neurons (13).

Although the effects of acute administration of kisspeptins on gonadotropin secretion have been described, the role that endogenous kisspeptins play in this process has not been determined, and it is not known whether there are other physiological roles for this ligand. It is important to establish the function of endogenous neurotransmitters because exogenously administered neurotransmitters do not always indicate what happens at the physiological level in a whole animal. As a step toward understanding the physiological role of endogenous kisspeptins, we have generated and characterized Kiss1-null mutant mice.

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Acknowledgments

This work was partly funded by a Biotechnology and Biological Sciences Research Council Industrial Partnership Award with Paradigm Therapeutics (BB/C003861/1). S.A.J.R.A. is supported by a Canada Research Chair in Molecular Oncology. W.H.C. is supported by the Ford Physiology Fund.

Acknowledgments

Abbreviations

IRES	internal ribosome entry site
FSH	follicle-stimulating hormone
LH	luteinizing hormone
AVPV	anteroventral periventricular nucleus
ARC	arcuate nucleus
GnRH	gonadotropin-releasing hormone.

Abbreviations

Footnotes

The authors declare no conflict of interest.

This article contains supporting information online at www.pnas.org/cgi/content/full/0704114104/DC1.

Footnotes

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