H-Y Antigen in the Teleost

H-Y antigen, presumably the product of mammalian testis-determining genes, has been detected in three species of teleost fish, Xiphophorus maculatus, Haplochromis burtoni , and Oryzias latipes , and in hybrids of the genus Tilapia . In X. maculatus H-Y was most readily detected in YY males, suggesting that a genetic determinant of H-Y antigen expression may exist on the teleost Y-chromosome. Although H-Y was detected in males and not in females in each of the species that we studied, male heterogamety has not been firmly established for H. burtoni . Thus despite the extreme phlyogenetic conservation of H-Y genes and their association with the Y-chromosome, it remains open to question whether H-Y will always be found in the heterogametic sex, and whether serologically defined H-Y antigen plays any part in the differentiation of the teleost gonad.     

H-Y Antigen and Sex-Determination in Turtles

H-Y antigen was investigated in 14 turtle species belonging to five different families. In 13 species the female was typed as H-Y antigen positive, only in Chinemys reevesi was the male H-Y antigen positive. Since in all vertebrate species studied till now, the expression of H-Y antigen is strictly correlated with the heterogametic sex, it can be assumed that in turtles both types of sex determining mechanisms are realized, namely the ZZ/ZW-and the XX/XY mechanism; both mechanisms are realized in species belonging to one and the same turtle family. However, most turtle species seem to be endowed with a ZZ/ZW sex determining mechanism.     

生殖细胞及性腺移植

生殖细胞和性腺移植研究近年来已取得了突破性进展。这两项技术对于农业、医学及动物繁殖学的研究具有深远的意义和很大的应用价值。本文从同源移植、异源移植、移植技术及其它移植相关问题等方面对生殖细胞和性腺移植进行了简要介绍,并阐述了近年来在这方面所取得的进展。     

Evidence for an H-Y Crossreactive Antigen in Invertebrates

A sex specific antigen which crossreacts with the mammalian H-Y antigen has been identified on the cell surface of hemocytes from the lobster ( Homarus americanus ) and the gonadal cells of three insect species. The hemocytes from the male lobster, the testicular cells from the male beetle ( P. cornutus ), and the ovarian cells from two Orthopteran species ( L. maderae and D. punctata ) specifically absorbed H-Y antibodies. The specificity of H-Y antibody absorptions by cells from only one sex, suggest that an ancestral H-Y-like antigen may be present in invertebrates which could be engaged in sexual (cellular) recognition events.     

Gonadal agenesis in a phenotypically normal female with positive H-Y antigen

Summary In a girl with primary amenorrhea, born from a consanguineous marriage, bilateral absence of gonads was established histologically. Cytogenetic studies demonstrated a 46,XY karyotype, and H-Y antigen determination was positive. In contrast with most reported cases of XY females with gonadal agenesis, normal development of female internal and external genitalia was present. Clinical and endocrinological features are reported, and the possible basis for the malformation is discussed.This work was supported by CNR, Centro di Studio Immunogenetica ed istocompatibilità, Torino, Italy     

Improved Transfection Efficiency of Chicken Gonadal Primordial Germ Cells for the Production of Transgenic Poultry

Electroporation is a common method of DNA transfection for many types of eukaryotic cells, but has not been attempted in avian primordial germ cells (PGCs). DNA uptake in chicken primordial germ cells (PGCs) was tested using electroporation with and without dimethyl sulfoxide (DMSO). Gonadal tissue and chicken embryonic fibroblasts (CEFs) were isolated from 6-day-old embryos (stage 29), transfected with pCMV carrying the bacterial lacZ gene, and cultured for 24 h. Gonadal primordial germ cells (gPGCs) were purified from culture using a Ficoll gradient. The addition of DMSO significantly increased the transfection efficiency of gPGCs but had no effect on chicken embryonic fibroblasts. Electroporation of gPGCs resulted in an 80% transfection efficiency, compared with about 17% observed with liposomes. Approximately 200 transfected gPGCs were injected into 2.5-day-old (stage 17) recipient embryos and the eggs were incubated for an additional 3.5 days, 7.5 days or to ...     

Dissecting Cell Lineage Specification and Sex Fate Determination in Gonadal Somatic Cells Using Single-Cell Transcriptomics

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Appearance of Primordial Germ Cells in Young Chick Blastoderms Cultured in vitro

Appearance of primordial germ cells (PGCs) in young chick blastoderms was investigated by the cultivation of only the epiblast or hypoblast. Presumptive PGCs exist in the epiblast before primitive-streak formation. They translocate gradually to the lower layer during early stages of primitive-streak formation, though substantial number of presumptive PGCs remain in the upper layer. The existing primary hypoblast under the epiblast is dispensable for the further differentiation of the PGCs.     

Ultrastructural Evidence that Chick Primordial Germ Cells Leave the Blood-Vascular System Prior to Migrating to the Gonadal Anlagen

It is known that chick primordial germ cells (PGCs), after separation from the endoderm in early embryonic development, temporarily circulate via the blood-vascular system and eventually migrate to the gonadal anlagen. However, direct evidence that circulating PGCs leave the blood vessels is lacking. The purpose of present study is to describe the ultrastructural features of PGCs as they emerge from the blood vessels. PGCs leaving the blood vessels were first examined with semi-thin sections stained with toluidine blue. Then, some of the sections were re-embedded in Epon 812, and sectioned for electron microscopy. PGCs were observed emerging from the capillaries in the region posterior to the omphalomesenteric arteries of the embryo, between the splanchnic mesoderm and open-gut endoderm, at stages 15–18 (about 2.5 days of incubation). Ultrastructurally, PGCs exhibited the protruding, bulge-like cytoplasmic processes through the endothelial gaps in the capillary walls. Prior to emerging, intravascular PGCs seemed to stick to the endothelium of the blood vessels. Thus, our results offer ultrastructural evidence that the circulating PGCs exit the blood vessels prior to migrating to the gonadal anlagen.     

Expression of H-Y Antigen in the Guppy (Lebistes reticulatus)

Expression of a mammalian cross-reactive H-Y antigen on the surface of cells derived from the male guppy ( Lebistes reticulatus ) is demonstrated. This finding further establishes the evolutionary conservation of H-Y antigen among lower vertebrates and provides a basis for speculation on the possible evolutionary association between H-Y antigen and sex determination.     

H-Y Antigen Expression in Temperature Sex-Reversed Turtles (Emys orbicularis)

H-Y antigen has been used as a marker for the heterogametic sex and is assumed to be an organizing factor for the heterogametic gonad. In the turtle Emys orbicularis , H-Y antigen is restricted to the female cells, indicating a female heterogamety (ZZ/ZW) sex-determining mechanism. Moreover, the sexual differentiation of the gonads is temperature sensitive, and complete sex reversal can be obtained at will. In this framework the relationships between H-Y antigen, temperature, and gonadal phenotype were studied. Mouse H-Y antiserum was absorbed with blood and gonadal cells of control wild male and female adults, and with blood and gonadal cells from three lots of young turtles from eggs incubated at 25–26°C (100% phenotypic males), at 30–30.5°C (100% phenotypic females), or at 28.5–29°C (majority of females with some males and intersexes). The residual activity of H-Y antiserum was then estimated using an immunobacterial rosette technique. In adults, both blood cells and gonadal cells were typed as H-Y negative in males and as H-Y positive in females. In each of the three lots of young, blood cells were H-Y negative in some individuals and H-Y positive in others. The proposed interpretation is that the H-Y negative individuals were genotypic males (ZZ) and the H-Y positive were genotypic females (ZW). The gonads of these animals were then pooled in different sets according to their sexual phenotype and to the presumed genotypic sex (i.e., blood H-Y phenotype). Testicular cells were typed as H-Y negative in genotypic males as well as in the presumed sex-reversed genotypic females; likewise, ovarian cells were typed as H-Y positive in genotypic females as well as in the presumed sex-reversed genotypic males. These results provide additional evidence that H-Y antigen expression is closely associated with ovarian structure in vertebrates displaying a ZZ/ZW sex-determining mechanism.     

Effects of H-Y Antigen on Morphologic and Endocrine Differentiation of Gonads in Mammals

The effects of H-Y antigen, released in a soluble state by male human Burkitt lymphomas and mouse teratomas, were studied in the fetal gonads from 13 to 21-day-old rats and 98-day-old calves, using histologic techniques, radioimmunoassays for testosterone, and bioassays for Müllerian-inhibiting substance (MIS). Whereas no effect was detected in the younger gonads, some structural changes were observed in bovine and 21-day-old rat ovaries when cultured for 5 days with Daudi and male teratoma supernatants, but there was no synthesis of testosterone or MIS. These observations raise questions concerning the critial period of sensitively to H-Y antigen of female gonads, and the dissociation between morphologic and biochemical events.     

Induction of H-Y Antigen in the Gonads of Male Quail Embryos by Diethylstilbestrol

In quails, H-Y antigen is induced by oestrogens in the gonads of the originally H-Y negative homogametic sex but not in non-gonadal tissues. This is consistent with the view that oestrogens act via H-Y antigen in the organization of the avian gonad.     

Gonadal Steroid Hormone Receptors and Sex Differences in the Hypothalamo-Pituitary-Adrenal Axis

The rapid activation of stress-responsive neuroendocrine systems is a basic reaction of animals to perturbations in their environment. One well-established response is that of the hypothalamo-pituitary-adrenal (HPA) axis. In rats, corticosterone is the major adrenal steroid secreted and is released in direct response to adrenocorticotropin (ACTH) secreted from the anterior pituitary gland. ACTH in turn is regulated by the hypothalamic factor, corticotropin-releasing hormone. A sex difference exists in the response of the HPA axis to stress, with females reacting more robustly than males. It has been demonstrated that in both sexes, products of the HPA axis inhibit reproductive function. Conversely, the sex differences in HPA function are in part due to differences in the circulating gonadal steroid hormone milieu. It appears that testosterone can act to inhibit HPA function, whereas estrogen can enhance HPA function. One mechanism by which androgens and estrogens modulate stress responses is through the binding to their cognate receptors in the central nervous system. The distribution and regulation of androgen and estrogen receptors within the CNS suggest possible sites and mechanisms by which gonadal steroid hormones can influence stress responses. In the case of androgens, data suggest that the control of the hypothalamic paraventricular nucleus is mediated trans-synaptically. For estrogen, modulation of the HPA axis may be due to changes in glucocorticoid receptor-mediated negative feedback mechanisms. The results of a variety of studies suggest that gonadal steroid hormones, particularly testosterone, modulate HPA activity in an attempt to prevent the deleterious effects of HPA activation on reproductive function.