Early aspects of sex differentiation in the gonads of chick embryos

In chick embryos whose sex had been previously identified by cytokaryologic methods, a light-microscope study of the number and dimensions of the germ cells (GCs) has been made from 2 to 7 days of incubation. Early differences between the sexes have been found. In females the GCs were larger and increased in number earlier than in males. This suggests an earlier differentiation of GCs in females. On the other hand, ultrastructural observations on GCs at 70 h incubation (colonization stage of the genital ridges) have revealed that male and female GCs differ from each other mainly in the amount of rough and smooth endoplasmic reticulum, mitochondria, glycogen particles and lipid droplets. This suggests early morpho-functional differences between male and female GCs.     

Tamoxifen and sex differentiation of the gonads in chick embryo

Tamoxifen or 4-hydroxytamoxifen were injected either alone or in combination with oestradiol into 4-5 day-old chick embryos in order to study their action on the sex differentiation of the gonads. The results of the histological study of the gonads performed at the stage of 16-19 days warrant the following conclusions: None of both anti-oestrogens exerts an effect on the testes. None of both compounds modifies the sex differentiation of the female gonads. Tamoxifen exerts an antagonistic action on the feminization of the testes by oestradiol. These conclusions do not lend support to the hypothesis according to which oestrogens play a role in normal sex differentiation of the female gonads.     

Lectin-binding carbohydrates in sexual differentiation of rat male and female gonads

Summary Development and sexual differentiation of the mammalian gonad involve changes in the type and distribution of different proteins and glycoproteins in and around the epithelial gonadal cords, the future seminiferous tubules in the testis, and follicles in the ovary. To study changes in cellular carbohydrate-containing compounds in the sex-specific morphogenesis of rat gonads, sections from embryonic, fetal and early postnatal gonads were labelled with seven different fluorescein isothiocyanate (FITC)-conjugated plant lectins of various carbohydrate-binding specificities. Double labelling of laminin with tetramethylrhodamine isothiocyanate (TRITC)-conjugated antibodies was used to outline the epithelial tissues. From the results we conclude that the abundance and similar distribution of carbohydrates in the early gonads of both sexes supports their sexually indifferent nature. Furthermore, the basement membranes of the differentiating gonadal cords in both sexes have common features, which differ, however, from those of the other developing urogenital organs. Changes in carbohydrate composition appear with the sexual differentiation of the gonads; the similarity of the changes in lectin binding to the gonadal cords of embryonic and fetal male, and to postnatal female, suggests similar mechanisms of cell-cell interactions in both sexes although activated at different developmental stages. These carbohydrate specificites at the tissue level should be taken into account together with cell-type specific changes, e.g. in the formation of the zona pellucida, when the phenomenon of polymorphic expression of different compounds is integrated into theories of epithelial differentiation.     

Lectin-binding carbohydrates in sexual differentiation of rat male and female gonads.

Development and sexual differentiation of the mammalian gonad involve changes in the type and distribution of different proteins and glycoproteins in and around the epithelial gonadal cords, the future seminiferous tubules in the testis, and follicles in the ovary. To study changes in cellular carbohydrate-containing compounds in the sex-specific morphogenesis of rat gonads, sections from embryonic, fetal and early postnatal gonads were labelled with seven different fluorescein isothiocyanate (FITC)-conjugated plant lectins of various carbohydrate-binding specificities. Double labelling of laminin with tetramethylrhodamine isothiocyanate (TRITC)-conjugated antibodies was used to outline the epithelial tissues. From the results we conclude that the abundance and similar distribution of carbohydrates in the early gonads of both sexes supports their sexually indifferent nature. Furthermore, the basement membranes of the differentiating gonadal cords in both sexes have common features, which differ, however, from those of the other developing urogenital organs. Changes in carbohydrate composition appear with the sexual differentiation of the gonads; the similarity of the changes in lectin binding to the gonadal cords of embryonic and fetal male, and to postnatal female, suggests similar mechanisms of cell-cell interactions in both sexes although activated at different developmental stages. These carbohydrate specificities at the tissue level should be taken into account together with cell-type specific changes, e.g. in the formation of the zona pellucida, when the phenomenon of polymorphic expression of different compounds is integrated into theories of epithelial differentiation.     

Estrogen target sites in the cloacal region of female and male chick embryos

Summary After intravenous injection of ³H-estradiol in the 12-day old chick embryo, radioactivity is concentrated in nuclei of certain cells in the cloacal area. The nuclear labeling is observed in mesenchymal cells along the different portions of the cloaca, and in an unidentified tissue located laterally to the cloaca. The labeled mesenchymal cells display a definite pattern of distribution along the epithelial wall of the cloaca, identical both in male and in female embryos. In the adjacent bursa of Fabricius, cells do not concentrate labeled hormone in their nuclei. The presence of estrogen receptors in the cloacal area of embryos of either sex adds evidence, at the cellular level, to support the concept of a neutral, or undifferentiated, sex with estradiol inhibiting this neutral male differentiation.Visiting scientistSupported by a Fellowship from the Délégation Générale à la Recherche scientifique et technique, Paris, France     

Effects of steroid sex hormones on chick embryo gonads in organ culture, with special reference to hormonal control of gonadal sex differentiation

At the initial stages of sex differentiation (7.5 and 8.5 days of incubation), chick embryo gonads were treated directly with testosterone or estradiol-17 beta in organ cultures. Chemically-defined media containing cholesterol as a steroid precursor were used. The differentiation of gonads in the 10 to 12-day controls, cultured in media containing no hormones, was close to that of gonads of equivalent age in ovo. Testosterone added to the medium exerted an inhibitory effect on the cortex of the female gonad and a masculinizing one on its medulla. The results of estradiol treatment confirmed the known feminizing effect of that hormone on the male gonad, the meiotic prophase in the genetically male germ cells being initiated in the induced cortex. These data may be interpreted in favour of a bihormonal theory of gonadal sex differentiation in birds, where the predominantly-synthesized male or female hormone in the gonad determines the male or female pattern of development of the corresponding gonad.     

Correlation between X-chromosome inactivation and cell differentiation in female preimplantation mouse embryos

By means of a cytological method involving BrdU incorporation and acridine orange fluorescence staining in combination with embryo manipulation, we studied X-chromosome activity in female preimplantation mouse embryos with special reference to the correlation between X-chromosome inactivation and cell differentiation. There was no sign of asynchronous replication between the two X chromosomes from the one-cell to intermediate blastocyst stage. The allocyclic X chromosome, first detected in late blastocysts, was paternal in origin, mostly replicating early in the S phase and limited to the trophectoderm. Subsequent X-chromosome inactivation occurring in the primary endoderm was also characterized by the involvement of the paternal X and early replication. Both X chromosomes continued to replicate synchronously in the embryonic ectoderm or epiblast at this stage. It was evident that overt cell differentiation preceded the appearance of the asynchronously replicating X chromosome in the trophectoderm and primary endoderm. This finding seems to support the view that cell differentiation is an important correlate of X-chromosome inactivation.     

Immunoreactive inhibin in plasma, amniotic fluid, and gonadal tissue of male and female chick embryos.

Immunoreactive inhibin was measured in plasma, amniotic fluid, gonads, and Wolffian bodies (mesonephros) of male and female chick embryos during the last week of their 21-day incubation period. The antiserum used was raised against bovine 31-kDa inhibin and was validated for RIA of inhibin in the chicken. Amniotic fluid concentrations of immunoreactive inhibin were relatively low and remained constant between Days 14 and 19. Plasma concentrations, in contrast, were high on Day 14 but declined steeply thereafter. Significantly higher plasma concentrations were noted in male than in female embryos and an even more pronounced sex difference was observed for the gonadal inhibin content. On Day 21, testes contained approximately 35 times more immunoreactive inhibin than ovaries. Surprisingly, inhibin contents in testes and male Wolffian bodies increased rather than decreased towards the end of the incubation period, indicating that gonadal and plasma inhibin concentrations are regulated, at least in part, independently. It is concluded that the chick embryo presents a convenient model for study of the secretion, the control, and the role of inhibin from fetal origin. The sex difference in plasma and gonadal inhibin suggests a differential role of inhibin in the development of the reproductive system of both sexes.