Sexual differentiation in three unconventional mammals: spotted hyenas, elephants and tammar wallabies.

The present review explores sexual differentiation in three non-conventional species: the spotted hyena, the elephant and the tammar wallaby, selected because of the natural challenges they present for contemporary understanding of sexual differentiation. According to the prevailing view of mammalian sexual differentiation, originally proposed by Alfred Jost, secretion of androgen and anti-Mullerian hormone (AMH) by the fetal testes during critical stages of development accounts for the full range of sexually dimorphic urogenital traits observed at birth. Jost's concept was subsequently expanded to encompass sexual differentiation of the brain and behavior. Although the central focus of this review involves urogenital development, we assume that the novel mechanisms described in this article have potentially significant implications for sexual differentiation of brain and behavior, a transposition with precedent in the history of this field. Contrary to the "specific" requirements of Jost's formulation, female spotted hyenas and elephants initially develop male-type external genitalia prior to gonadal differentiation. In addition, the administration of anti-androgens to pregnant female spotted hyenas does not prevent the formation of a scrotum, pseudoscrotum, penis or penile clitoris in the offspring of treated females, although it is not yet clear whether the creation of masculine genitalia involves other steroids or whether there is a genetic mechanism bypassing a hormonal mediator. Wallabies, where sexual differentiation occurs in the pouch after birth, provide the most conclusive evidence for direct genetic control of sexual dimorphism, with the scrotum developing only in males and the pouch and mammary glands only in females, before differentiation of the gonads. The development of the pouch and mammary gland in females and the scrotum in males is controlled by genes on the X chromosome. In keeping with the "expanded" version of Jost's formulation, secretion of androgens by the fetal testes provides the best current account of a broad array of sex differences in reproductive morphology and endocrinology of the spotted hyena, and androgens are essential for development of the prostate and penis of the wallaby. But the essential circulating androgen in the male wallaby is 5alpha androstanediol, locally converted in target tissues to DHT, while in the pregnant female hyena, androstenedione, secreted by the maternal ovary, is converted by the placenta to testosterone (and estradiol) and transferred to the developing fetus. Testicular testosterone certainly seems to be responsible for the behavioral phenomenon of musth in male elephants. Both spotted hyenas and elephants display matrilineal social organization, and, in both species, female genital morphology requires feminine cooperation for successful copulation. We conclude that not all aspects of sexual differentiation have been delegated to testicular hormones in these mammals. In addition, we suggest that research on urogenital development in these non-traditional species directs attention to processes that may well be operating during the sexual differentiation of morphology and behavior in more common laboratory mammals, albeit in less dramatic fashion.

Metabolic assessment of wallaby blastocysts during embryonic diapause and subsequent reactivation.

The metabolism of tammar wallaby (Macropus eugenii) blastocysts was analysed by means of quantitative fluorescence microscopy during embryonic diapause and 2, 3, 4, 5, 8 and 10 days after reactivation to determine nutrient preferences during metabolic reactivation of the blastocyst. The surface area of quiescent blastocysts was 0.16 +/- 0.02 mm2 (mean +/- s.e.m.), and increased to 0.44 +/- 0.04 mm2 (P < 0.05) by Day 8 after removal of the sucking stimulus of the pouch young (RPY). Day-10 blastocysts, analysed over two successive breeding seasons, were significantly different in size from each other (Group A, 1992: 4.44 +/- 1.47 mm2; Group B, 1993: 18.87 +/- 4.62 mm2; P < 0.01), and both groups were significantly different in size from diapausing blastocysts (P < 0.01). There was no significant difference in carbohydrate uptake or production by blastocysts during the first five days after RPY. Glucose uptake by blastocysts recovered 8 days after RPY (61.9 +/- 30.0 pmol embryo-1 h-1) was significantly greater than that by Day-0 blastocysts (17.9 +/- 5.5 pmol embryo-1 h-1) and glucose uptake by both groups of Day-10 blastocysts (Group A, 174.0 +/- 28.4 pmol embryo-1 h-1; Group B, 616.0 +/- 239.0 pmol embryo-1 h-1) was significantly different from that by Day-0 blastocysts (P < 0.01). Pyruvate uptake by Day-10 blastocysts (Group A, 46.0 +/- 32.2 pmol embryo-1 h-1; Group B, 250.0 +/- 136.0 pmol embryo-1 h-1; P < 0.01) increased significantly compared with that by Day-0 blastocysts (6.4 +/- 1.6 pmol embryo-1 h-1; P < 0.01). Lactate production by Day-10 blastocysts (Group A, 186.7 +/- 30.3 pmol embryo-1 h-1; Group B, 285 +/- 129 pmol embryo-1 h-1; P > 0.01) was also significantly different from that by quiescent blastocysts (41.20 +/- 9.6 pmol embryo-1 h-1). There was a linear relationship between surface area and glucose uptake and surface area and pyruvate uptake (r2 = 0.965 and r2 = 0.971 respectively). Despite increases in carbohydrate uptake, there was a proportional decrease in lactate production indicating an increase in oxidative metabolism during reactivation. This suggests that there may be a metabolic switch at, or around, Day 5 after RPY.