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Marsupial Anti-Müllerian Hormone Gene Structure, Regulatory Elements, and Expression¹

Department of Zoology,⁴ University of Melbourne, Victoria 3010, Australia Department of Molecular Genetics,⁵ University of Texas M. D. Anderson Cancer Center, Houston, Texas 77030 Comparative Genomics Research Group,⁶ Research School of Biological Sciences, Australian National University, Canberra ACT 2601, Australia

During male sexual development in reptiles, birds, and mammals, anti-Müllerian hormone (AMH) induces the regression of the Müllerian ducts that normally form the primordia of the female reproductive tract. Whereas Müllerian duct regression occurs during fetal development in eutherian mammals, in marsupial mammals this process occurs after birth. To investigate AMH in a marsupial, we isolated an orthologue from the tammar wallaby (Macropus eugenii) and characterized its expression in the testes and ovaries during development. The wallaby AMH gene is highly conserved with the eutherian orthologues that have been studied, particularly within the encoded C-terminal mature domain. The N-terminus of marsupial AMH is divergent and larger than that of eutherian species. It is located on chromosome 3/4, consistent with its autosomal localization in other species. The wallaby 5' regulatory region, like eutherian AMH genes, contains binding sites for SF1, SOX9, and GATA factors but also contains a putative SRY-binding site. AMH expression in the developing testis begins at the time of seminiferous cord formation at 2 days post partum, and Müllerian duct regression begins shortly afterward. In the developing testis, AMH is localized in the cytoplasm of the Sertoli cells but is lost by adulthood. In the developing ovary, there is no detectable AMH expression, but in adults it is produced by the granulosa cells of primary and secondary follicles. It is not detectable in atretic follicles. Collectively, these studies suggest that AMH expression has been conserved during mammalian evolution and is intimately linked to upstream sex determination mechanisms.

¹ Supported by a grant from the National Institutes of Health (NIH) HD30284 to R.R.B. and the National Health and Medical Research Council of Australia (NHMRC) grant 940658 to M.B.R. and G.S.; A.J.P. was supported by a C.J. Martin Research Fellowship from the NHMRC; DNA sequencing was supported by NIH Cancer Center Support grant CA16672; A.J.P. and D.J.W. contributed equally to this work; M.B.R. and R.R.B. are co-senior authors.

² Correspondence: Richard R. Behringer, Department of Molecular Genetics, University of Texas M. D. Anderson Cancer Center, 1515 Holcombe Blvd., Houston, TX 77030. FAX: 713 794 4394; rrb{at}mdanderson.org

³ Current address: Pfizer Kalamazoo Laboratories, Pfizer Inc., Kalamazoo, MI 49007

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