Reproduction, Fertility and Development Reproduction, Fertility and Development Society
Vertebrate reproductive science and technology

Formation of 5α-reduced androgens in the testes and urogenital tract of the grey short-tailed opossum, Monodelphis domestica

Jean D. Wilson A B C , Marilyn B. Renfree A , Richard J. Auchus B , Andrew J. Pask A and Geoffrey Shaw A
+ Author Affiliations
- Author Affiliations

A Department of Zoology, University of Melbourne, Vic. 3010, Australia.

B Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, TX 75390-8857, USA.

C Corresponding author. Email:

Reproduction, Fertility and Development 21(5) 649-654
Submitted: 10 November 2008  Accepted: 20 March 2009   Published: 21 May 2009


Testicular 5α-reduced androgens, largely 5α-androstane-3α,17β-diol (androstanediol), are responsible for virilisation of pouch young in one marsupial (the tammar wallaby), but are not formed until later in development in another marsupial (the brushtail possum) and in rodents. Because the mechanism of virilisation of the urogenital tract in the grey short-tailed opossum Monodelphis domestica has never been defined, androgen formation and metabolism were investigated in this species. Testis fragments from grey short-tailed opossums of a wide range of ages were incubated with [3H]-progesterone and the metabolites were separated by high-performance liquid chromatography (HPLC). The only 19-carbon metabolites identified in the youngest ages (5–26 days) and the major metabolites in adult testes were testosterone and androstenedione. At 30, 42 and 49 days of age, dihydrotestosterone and small amounts of androstanediol were present. Time-sequence studies indicated that dihydrotestosterone and androstanediol were formed from the 5α-reduction (and 3-keto reduction) of testosterone. In a second series of experiments, tissue fragments of a variety of urogenital tract tissues were incubated with [3H]-testosterone and the metabolites separated by HPLC. During the interval in which male urogenital tract differentiation takes place in this species (between Days 15 and 28), the major metabolite identified was dihydrotestosterone. We conclude that the timing of 5α-reductase expression in the testes of the grey short-tailed possum resembles that of rodents and the brushtail possum rather than that of the tammar wallaby and that dihydrotestosterone is probably the intracellular androgen responsible for virilisation of the urogenital tract in this species.


This study was supported by grant 208911 from the National Health and Medical Research Council of Australia (NHMRC) and by Grant I-1493 from the Robert A. Welsh Foundation. A.J.P. was supported by an NHMRC RD Wright Fellowship and M.B.R. was supported by an Australian Research Council Federation Fellowship. The authors thank C. David Vance for performing the HPLC of the samples and Professor Norman Saunders and Dr Joakim Ek for providing the opossums.


Chase, D. J. , and Payne, A. H. (1983). Changes in Leydig cell function during sexual maturation in the mouse. Biol. Reprod. 29, 1194–1200.
CrossRef | CAS | PubMed |

Fadem, B. H. , and Harder, J. D. (1992). Evidence for high levels of androgen in peripheral plasma during postnatal development in a marsupial: The gray short-tailed opossum (Monodelphis domestica). Biol. Reprod. 46, 105–108.
CrossRef | CAS | PubMed |

Frungieri, M. B. , Gonzalez-Calvar, S. I. , Bartke, A. , and Calandra, R. S. (1999). Influence of age and photoperiod on steroidogenic function of the testis of the golden hamster. Int. J. Androl. 22, 243–252.
CrossRef | CAS | PubMed |

Ge, R.-S. , and Hardy, M. P. (1998). Variation in the end products of androgen biosynthesis and metabolism during postnatal differentiation of rat Leydig cells. Endocrinology 139, 3787–3795.
CrossRef | CAS | PubMed |

Ge, R.-S. , Hardy, D. O. , Catterall, J. F. , and Hardy, M. P. (1999). Opposing changes in 3α-hydroxysteroid dehydrogenase oxidative and reductive activities in rat Leydig cells during pubertal development. Biol. Reprod. 60, 855–860.
CrossRef | CAS | PubMed |

Gloyna, R. E. , and Wilson, J. D. (1969). A comparative study of the conversion of testosterone to 17β-hydroxy-5α-androstane-3-one (dihydrotestosterone) by prostate and epididymis. J. Clin. Endocrinol. Metab. 29, 970–977.
CAS | PubMed |

Leihy, M. W. , Shaw, G. , Wilson, J. D. , and Renfree, M. B. (2004). Penile development is initiated in the tammar wallaby pouch young during the period when 5α-androstane-3α,17β-diol is secreted by the testes. Endocrinology 145, 3346–3352.
CrossRef | CAS | PubMed |

Mackay, S. , Xie, Q. , Ullmann, S. L. , Gilmore, D. P. , and Payne, A. P. (2004). Postnatal development of the reproductive system in the grey short-tailed opossum, Monodelphis domestica. Anat. Embryol. 208, 121–133.
CrossRef | CAS | PubMed |

Mahendroo, M. , Wilson, J. D. , Richardson, J. A. , and Auchus, R. J. (2004). Steroid 5α-reductase 1 promotes 5α-androstane-3α,17β-diol synthesis in immature mouse testes by two pathways. Mol. Cell. Endocrinol. 222, 113–120.
CrossRef | CAS | PubMed |

R Development Core Team (2008). ‘R: A Language and Environment for Statistical Computing.’ (R Foundation for Statistical Computing, Vienna.) Available at

Shaw, G. , Renfree, M. B. , Leihy, M. W. , Shackleton, C. H. L. , Roitman, E. , and Wilson, J. D. (2000). Prostate formation in a marsupial is mediated by the testicular androgen 5α-androstane-3α,17β-diol. Proc. Natl Acad. Sci. USA 97, 12 256–12 259.
CrossRef | CAS |

Shaw, G. , Fenelon, J. , Sichlau, M. , Auchus, R. J. , Wilson, J. D. , and Renfree, M. B. (2006). Role of the alternate pathway of dihydrotestosterone formation in virilization of the wolffian ducts of the tammar wallaby, Macropus eugenii. Endocrinology 147, 2368–2373.
CrossRef | CAS | PubMed |

Sheffield, J. W. , and O’Shaughnessy, P. J. (1988). Testicular steroid metabolism during development in the normal and hypogonadal mouse. J. Endocrinol. 119, 257–264.
CrossRef | CAS | PubMed |

Wilson, J. D. , Auchus, R. J. , Leihy, M. W. , Guryev, O. L. , Estabrook, R. W. , Osborn, S. M. , Shaw, G. , and Renfree, M. B. (2003). 5α-Androstane-3α,17β-diol is formed in tammar wallaby pouch young by a pathway involving 5α-pregnane-3α,17α-diol-20-one as a key intermediate. Endocrinology 144, 575–580.
CrossRef | CAS | PubMed |

Wilson, J. D. , Shaw, G. , Renfree, M. B. , Auchus, R. J. , Leihy, M. W. , and Eckery, D. C. (2005). Ontogeny and pathway of formation of 5α-androstane-3α,17β-diol in the testes of the immature brushtail possum Tricosurus vulpecula. Reprod. Fertil. Dev. 17, 603–609.
CrossRef | CAS | PubMed |

Xie, Q. , Mackay, S. , Ullmann, S. L. , Gilmore, D. P. , Payne, A. P. , and Gray, C. (1998). Postnatal development of Leydig cells in the opossum (Monodelphis domestica). Biol. Reprod. 58, 664–669.
CrossRef | CAS | PubMed |

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