Genetics of mammalian sex determination: some unloved exceptions.

The genetics of sex determination is a child of the twentieth century, which overturned the previously held view that sex was determined by the environment. The last quarter of the century witnessed an active search for sex-determining genes in mammals. Although successful, the modus operandi of these genes remained unknown, and the relationship between the sex-determining systems of mammals and other vertebrates remained enigmatic. To overcome these problems, scientists in the 21st century should heed William Bateson's counsel to treasure exceptions, for they point the way to progress. One exception to conventional concepts of sex determination is the bilaterally asymmetrical distribution of ovaries and testes in true hermaphroditism. Ovaries favour the left side in humans and the right side in mice. Observations suggesting that a reversal of asymmetry may occur with increasing organ size may point to a possible explanation. A reevaluation is also required regarding the beginning of sex differentiation, in view of mounting evidence of a sex difference in growth rates of early embryos. Another question to be settled is whether the function of SRY is confined to the fetal gonad. The recent demonstration that Sry induces cell proliferation in the fetal mouse gonad (Schmahl et al., 2000) further emphasizes the importance of differential growth in sex determination and differentiation. It is suggested that SRY represents an additional growth-promoting gene sequestered by mammals to enable the XY embryo to undergo male sex differentiation in the female hormonal environment of the uterus. An increased awareness of the relationship between growth and gonadal differentiation should lead to a better understanding of sex determination in mammals and an ability to relate the function of sex-determining genes to the effects of environmental factors. J. Exp. Zool. 290:484-489, 2001.

Genetics of sex determination: exceptions that prove the rule.

The dogma that male and female embryos develop in identical fashion until SRY initiates Sertoli cell differentiation in the hitherto bipotential gonad is in need of reevaluation in the light of data that do not fit into this scheme. One of the exceptions that proves the rule of sex determination is true hermaphroditism, the existence of individuals with both testicular and ovarian tissue. Furthermore, the two types of tissues are asymmetrically distributed, ovaries being more common on the left side and testes and ovotestes on the right. Hermaphrodite mice also exhibit bilateral asymmetry of gonad differentiation, but in the opposite direction: ovaries on the right, testes and ovotestes on the left. To explain these asymmetries, it is necessary to consider the relationship between growth and gonadal differentiation. The idea that accelerated growth precedes histological differentiation of the testis has recently been confirmed by the finding that Sry induces cell proliferation in fetal mouse gonads, suggesting that the differentiation of Sertoli cells may be dependent on a critical cell number. Recent evidence has also shown that XY embryos develop faster than their XX counterparts at very early stages of development, and it has been reported that SRY and ZFY are expressed in early human and murine embryos. The relationship between growth and sex differentiation links the mammalian system with those of nonmammalian vertebrates with temperature-dependent sex determination. Early growth differences between male and female human embryos question the belief that all sex differences in later life are due to gonadal hormones.

Gonadal development and birth weight in X*X and X*Y females of the wood lemming, Myopus schisticolor.

A quantitative histological analysis of ovaries from 8- to 10-day-old wood lemmings revealed significant differences between females with X*Y and X*X sex chromosome constitutions. The ovarian volume of X*Y females was on average 57% of X*X, and the number of oocytes was less than half in X*Y compared to X*X. However, the frequency of growing oocytes in relation to the total number was 6.5% for X*Y compared to 3.0% for X*X. Oogenesis in X*Y wood lemmings resembles in many respects that of mice heterozygous for certain translocations and with tertiary trisomy (Ts31H), and those with X0 monosomy. The fertility in X*Y wood lemmings is not reduced. On the contrary, X*Y females have a higher reproductive fitness than X*X and XX. This is discussed in relation to the present findings. The body weight at birth was 8% higher in X*Y than in X*X.

Three thousand years of questioning sex determination.

Of all the inborn differences that distinguish individual humans, as well as other animals, sex exerts the most far-reaching effects, and the question, what determines it, has been debated throughout history. A discriminating reading of Biblical and Ancient Greek sources reveals surprising insights that are relevant to present-day biology. The material basis of generation was inaccessible until, following the invention of the microscope and the discovery of "spermatic animalcules" in the 17th century, the 19th century witnessed the discovery of the mammalian egg, the nature of sperm, and the process of fertilization. Sex was thought to be determined by external conditions. The 20th century developed the genetics of sex determination. The search for the mammalian testis-determining gene during the last quarter century culminated in the discovery of SRY, soon to be accompanied by non-Y chromosome sex- determining genes. During the same period, data accumulated that testicular differentiation was accompanied by accelerated gonadal growth; subsequently, differences in growth were shown to distinguish early XX from early XY embryos. Other research showed that temperature-dependent sex determination was widely distributed among reptiles, thus illustrating that the mammalian system of sex determination is of recent evolutionary origin, adopted in response to homoiothermy and placentation. The recent discovery that Sry induces cell proliferation in the gonads of fetal mice suggests that the task for the 21st century will be to aim beyond simple genotype/phenotype correlations by unraveling the relationship between genes and epigenetic factors acting on cell growth during development and affecting the phenotype in later life.

Phenotypic manifestations during the development of the dominant and default gonads in mammals and birds.

The dominant embryonic gonad--testis in mammals and ovary in birds--secretes one or more morphogenetic substances that exert a major effect on the phenotype of the embryo. When deprived of their gonads, mammalian embryos develop into females, and avian embryos assume predominantly male characteristics, although retaining both oviducts. In order to fulfill their task of masculinizing the reproductive tract, mammalian testes grow and differentiate faster than ovaries. In birds the pattern is less straightforward. In 5-day-old embryos of White Leghorn chickens, sexual differentiation manifests itself in two different ways: (1) the gonads of ZZ embryos are larger, and on day 6 contain more protein and DNA than those of ZW embryos; (2) in both sexes, left gonads are larger than right gonads and contain a thick "germinal epithelium" capable of giving rise to an ovarian cortex under the influence of oestrogen. The pattern changes in embryos aged between 7 and 8 days, when the left gonad of ZW embryos outgrows all others, developing into an ovary, and when the bilateral asymmetry between left and right gonads increases in female embryos. A remnant of gonadal bilateral asymmetry is seen in the distribution of gonads in cases of true hermaphroditism in humans and other mammals. Whereas the initial fast growth of the mammalian testis is assumed to be due to one or more Y-chromosomal genes, that of the early avian testis is mostly simply explained as the effect in the disomic state of one or more genes on the Z chromosome. However, the later growth of the avian ovary is more likely to be due to oestrogen than to a direct gene effect. It is postulated that oestrogen has lost its power to determine ovarian development in mammals, in which both sexes are exposed to the oestrogen-rich environment of the uterus. Hence, the task of sex determination devolves on the fetal testis, whose early development and hormonal function are required to induce the male phenotype, the female phenotype arising in default mode.

Sex-determining mechanisms in animals.

Biological mechanisms leading to the development of males and females are extremely varied. In the XX/XY system, the male has an unequal pair of chromosomes, while in the ZZ/ZW system, the unequal pair is in the female. Sex can also be determined by the temperature of incubation. Recent research has focused on the identification of sex-determining genes, culminating in the demonstration that the Sry gene on the Y chromosome of mice can induce male development in genetically female XX mouse embryos. Nevertheless, the occurrence of phenotypes in apparent contrast to the genotype suggests that the genetic male/female switch is not simple, and there may be common features linking different sex-determining mechanisms. There is increasing evidence that such a link may be provided by the induction of growth differences, and that the primary sex difference may result from the distinction between fast versus slow growth.

A quantitative investigation of gonadal feminization by diethylstilboestrol of genetically male embryos of the quail Coturnix coturnix japonica.

The effect of diethylstilboestrol on gonad development in quail embryos has been quantitatively analysed. Quail embryos at 4 days of incubation were treated with diethylstilboestrol (DES), using the egg dipping method. At 10 days of incubation, embryos were removed and killed by decapitation. Tissues were prepared for chromosome analysis, and the parts of the abdomen containing the gonads were prepared for serial sectioning and quantitative assessment. Left gonads of DES-treated male embryos resembled ovaries histologically, while their right gonads were markedly reduced in size. Right gonads of DES-treated female embryos were also further reduced by treatment with DES. There was no statistically significant effect by DES treatment on the size of left gonads, although the ratio of left compared with right gonadal volumes was highly significant. Since, in birds, the left embryonic gonad has ambisexual potential, while the potential of the right gonad is exclusively masculine, these results exemplify the adverse effect exerted by oestrogen on male sexual development in vertebrates.

Blastocysts prepare for the race to be male.

Recent findings in different mammalian species have demonstrated that XY embryos grow faster than XX embryos before the gonads are differentiated. In mice and cattle, accelerated development is already evident in XY blastocysts, while in the rat and in human fetuses a quantitative sex difference has been shown to be present before testicular differentiation has occurred. These data demonstrate that in these species the histological differentiation of the testis, which occurs early and rapidly, is preceded by an increased growth rate of the embryo. This may be expected to increase the probability of the gonad reaching the threshold for testis development, since it is known that developmental delay can result in ovarian differentiation. It is postulated that the fast development of the male may be an adaptation to the reproductive biology of eutherian mammals, in which development of both sexes occurs in the hormonal environment of the uterus. The question is raised as to a possible connection between sex-related growth and other sex differences, such as longevity.

Gonadal sex differentiation in embryos and neonates of the marsupial, Monodelphis domestica: arrest of testis development in postterm embryos.

Growth and histological differentiation were studied in 8 litters of embryos and 4 litters of neonate grey short-tailed opossums, Monodelphis domestica. The embryonic litters included 2 that had passed their expected birth date, and whose weights exceeded the usual birthweights; we refer to these litters as 'postmature'. There was an abrupt increase in the growth rate of XY gonads after birth, but this was not seen in XX gonads. Although there was evidence of testicular differentiation in XY gonads on the day before the expected birth, testicular differentiation was found to be blocked in postmature litters. The growth of XX gonads in postmature embryos was not affected. In view of evidence that exogenous oestrogens feminise the gonads of genetic males in some species of marsupials including Monodelphis domestica, the question arises whether oestrogen is responsible for the failure of testes to continue their development in utero. We suggest that the ability of functional testes to develop in the presence of oestrogen may be a fundamental requirement distinguishing eutherian mammals from other vertebrates, including marsupials.

Sex determination and sex reversal: genotype, phenotype, dogma and semantics.

The genetic terminology of sex determination and sex differentiation is examined in relation to its underlying biological basis. On the assumption that the function of the testis is to produce hormones and spermatozoa, the hypothesis of a single Y-chromosomal testis-determining gene with a dominant effect is shown to run counter to the following observed facts: a lowering in testosterone levels and an increase in the incidence of undescended testes, in addition to sterility, in males with multiple X chromosomes; abnormalities of the testes in autosomal trisomies; phenotypic abnormalities of XX males apparently increasing with decreasing amounts of Y-chromosomal material; the occurrence of patients with gonadal dysgenesis and XY males with ambiguous genitalia in the same sibship; the occurrence of identical SRY mutations in patients with gonadal dysgenesis and fertile males in the same pedigree; and the development of XY female and hermaphrodite mice having the same genetic constitution. The role of X inactivation in the production of males, females and hermaphrodites in T(X;16)16H mice has previously been suggested but not unequivocally demonstrated; moreover, X inactivation cannot account for the observed bilateral asymmetry of gonadal differentiation in XY hermaphrodites in humans and mice. There is evidence for a delay in development of the supporting cells in XY mice with ovarian formation. Once testicular differentiation and male hormone secretion have begun, other Y-chromosomal genes are required to maintain spermatogenesis and to complete spermiogenesis, but these genes do not function effectively in the presence of more than one X chromosome. The impairment of spermatogenesis by many other chromosome abnormalities seems to be more severe than that of oogenesis. It is concluded that the notion of a single testis-determining gene being responsible for male sex differentiation lacks biological validity, and that the genotype of a functional, i.e. fertile, male differs from that of a functional female by the presence of multiple Y-chromosomal genes in association with but a single X chromosome. Male sex differentiation in XY individuals can be further impaired by a euploid, but inappropriate, genetic background. The genes involved in testis development may function as growth regulators in the tissues in which they are active.

Unpaired chromosomes at meiosis: cause or effect of gametogenic insufficiency?

Pairing failure at meiosis has been postulated as a cause of gametogenic arrest in both heterozygous translocation carriers and males whose spermatocytes exhibit univalent X and Y chromosomes. The present investigation is a survey of pachytene translocation configurations, at the electron microscopic level, in six stocks of mice, comprising a total of 464 spermatocytes and 343 oocytes. Univalence of the X and Y chromosomes was studied in the same stocks, as well as in three additional homozygous translocation stocks. Fully paired as well as asynaptic configurations were found in all translocation stocks, and the proportions of each configuration differed considerably between spermatocytes and oocytes of mice carrying the same translocation. In both spermatocytes and oocytes, other pairing anomalies were more frequent in cells with asynapsed than with fully synapsed configurations, and spermatocytes with univalent sex chromosomes had a higher proportion of autosomal anomalies than did spermatocytes with XY bivalents. It is concluded that pairing failure at meiosis is primarily a symptom, rather than a cause, of gametogenic arrest, and that chromosome rearrangements, even if they appear to be balanced, may affect the rate of atresia by interfering with the normal rate of meiotic progression. Once pairing failure is established, it could secondarily increase the probability of gametogenic failure.