Premature ovarian failure in androgen receptor-deficient mice

Hiroko Shiina

Takahiro Matsumoto

Takashi Sato

Katsuhide Igarashi

Takashi Nakamura+5 authors

^{Institute of Molecular and Cellular Biosciences, University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-0032, Japan;}^{Exploratory Research for Advanced Technology, Japan Science and Technology, 4-1-8 Honcho, Kawaguchi, Saitama 332-0012, Japan;}^{Department of Obstetrics and Gynecology, Institute of Clinical Medicine, University of Tsukuba, 1-1-1 Tennoudai, Tsukuba, Ibaraki 305-8575, Japan;}^{Division of Cellular and Molecular Toxicology, National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan; and}^{Institut de Genetique et de Biologie Moleculaire et Cellulaire, Centre National de la Recherche Scientifique, Institut National de la Santé et de la Recherche Médicale, Universite Louis Pasteur, College de France, 67404 Illkirch, Strasbourg, France}

^{To whom correspondence should be addressed. E-mail:}pj.ca.oykot-u.cce.liam@otaksu.

^{H.S. and T.M. contributed equally to this work.}

Edited by Bert W. O'Malley, Baylor College of Medicine, Houston, TX, and approved November 10, 2005

Received 2005 Aug 5

Abstract

Premature ovarian failure (POF) syndrome, an early decline of ovarian function in women, is frequently associated with X chromosome abnormalities ranging from various Xq deletions to complete loss of one of the X chromosomes. However, the genetic locus responsible for the POF remains unknown, and no candidate gene has been identified. Using the Cre/LoxP system, we have disrupted the mouse X chromosome androgen receptor (Ar) gene. Female AR^{mice appeared normal but developed the POF phenotype with aberrant ovarian gene expression. Eight-week-old female}AR^{mice are fertile, but they have lower follicle numbers and impaired mammary development, and they produce only half of the normal number of pups per litter. Forty-week-old}AR^{mice are infertile because of complete loss of follicles. Genome-wide microarray analysis of mRNA from}AR^{ovaries revealed that a number of major regulators of folliculogenesis were under transcriptional control by AR. Our findings suggest that AR function is required for normal female reproduction, particularly folliculogenesis, and that AR is a potential therapeutic target in POF syndrome.}

Keywords: male hormone, nuclear receptor, female physiology, folliculogenesis, kit ligand

Abstract

Premature ovarian failure (POF) is defined as an early decline of ovarian function after seemingly normal folliculogenesis (1). Genetic causes of POF have been frequently associated with X chromosome abnormalities (1, 2). Complete loss of one of the X chromosomes, as in Turner syndrome, and various Xq deletions are commonly identified as a cause of POF. However, responsible X-linked genes and their downstream targets have not been identified so far.

The androgen receptor (Ar) gene, which is the only sex hormone receptor gene on the X chromosome, is well known to be essential not only for the male reproductive system, but also for male physiology. In contrast, androgens are considered as male hormones; therefore, little is known about androgens' actions in female physiology, although AR expression in growing follicles has been described (3). However, because excessive androgen production in polycystic ovary syndrome causes infertility with abnormal menstrual cycles (4, 5), it is possible that AR-mediated androgen signaling also plays an important physiological role in the female reproductive system. Recently, using Cre/LoxP system, we generated an AR-null mutant mouse line (6) and demonstrated that inactivation of AR resulted in arrest of testicular development and spermatogenesis, impaired brain masculinization, high-turnover osteopenia, and late onset of obesity in males (7–9). At the same time, no overt physical or growth abnormalities were observed in female AR^{mice. Therefore, to further examine potential role of AR in female physiology, we characterized female reproductive system in}AR^{females. Herein we show that female}AR^{mice develop the POF phenotype. At 3 weeks of age,}AR^{females had apparently normal ovaries with numbers of follicles similar to those in the wild-type females. However, thereafter the number of healthy follicles in the}AR^{ovary gradually declined, with a marked increase of atretic follicles, and by 40 weeks}AR^{mice became infertile, with no follicle detectable in the ovary. Reflecting this age-dependent progression in ovarian abnormality, several genes known to be involved in the oocyte–granulosa cell regulatory loop were identified by microarray analysis as AR downstream target genes. These findings clearly demonstrate that AR-mediated androgen signaling is indispensable for the maintenance of folliculogenesis and implicate impaired androgen signaling as a potential cause of the POF syndrome.}

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Acknowledgments

We thank T. Iwamori and H. Tojo for expert advice on mammary gland anatomy, Y. Kanai for ovarian phenotypic analysis, members of the KO project team at the laboratory of Nuclear Signaling (Institute of Molecular and Cellular Biosciences) for their support, A. P. Kouzmenko for helpful suggestions, and H. Higuchi for manuscript preparation. This work was supported in part by the Program for Promotion of Basic Research Activities for Innovative Biosciences and priority areas from the Ministry of Education, Culture, Sports, Science, and Technology (to S.K.).

Acknowledgments

Notes

Author contributions: J.K., H.Y., and S.K. designed research; H.S., T.M., T.S., K.I., J.M., S.T., M.S., I.T., and T.N. performed research; D.M. and P.C. contributed new reagents/analytic tools; H.S. analyzed data; and S.K. wrote the paper.

Conflict of interest statement: No conflicts declared.

This paper was submitted directly (Track II) to the PNAS office.

Abbreviations: AR, androgen receptor; DHT, 5α-dihydrotestosterone; POF, premature ovarian failure.

Notes

Conflict of interest statement: No conflicts declared.

This paper was submitted directly (Track II) to the PNAS office.

Abbreviations: AR, androgen receptor; DHT, 5α-dihydrotestosterone; POF, premature ovarian failure.

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