Sex differentiation of germ cells in the teleost, Oryzias latipes, during normal embryonic development

Development ◽  
1972 ◽  
Vol 28 (2) ◽  
pp. 385-395
Author(s):  
Noriyuki Satoh ◽  
Nobuo Egami

Mitotic and meiotic activities of germ cells during early development in the medaka, Oryzias latipes, are dealt with in this report. Primordial germ cells were obviously distinguishable from somatic cells 3 days after fertilization and began to proliferate about 7 days after fertilization. The mean number of primordial germ cells increased during a period of 7–10 days after fertilization, reaching about 90 immediately before hatching. Newly hatched fry could be classified into two types according to the number and the nucleic activity of germ cells in the gonadal rudiment. One type consisted of fry containing about 100 germ cells and no cells in the meiotic prophase. In the other type of fry the number of germ cells increased by mitotic divisions and some of the cells began to enter into the meiotic prophase. During the course of further development the fry of the former type differentiated into males and the latter into females. Therefore it can be concluded that the morphological sex differentiation of germ cells occurs at the time of hatching. However, no sexual differences in the histological structure of somatic elements in the gonad are observable at that time.

1953 ◽  
Vol 31 (2) ◽  
pp. 73-79 ◽  
Author(s):  
J. G. Robertson

The differentiation of the gonad is described in chum salmon embryos and alevins. Contrary to classical findings in teleosts, sex differentiation in the chum salmon proceeds in the male or female direction without an intermediate female phase. From an initially indifferent gonad there is a progressive development of one sex or the other. The organ forms as a fold from the splanchnic mesoderm and, at the time of first appearance, contains primordial germ cells. These enlarge to form the definitive germ cells which, after a series of divisions, form smaller oogonia or spermatogonia. Oogonia are followed by primary and secondary (growing) oocytes, the appearance of which is the criterion of sex distinction. Spermatogonia continue to multiply but do not undergo growth in the alevin. The ovary develops an open endovarial canal and is supported by a prominent mesovarium. The testis remains small and, in the alevin, develops no ducts. It is suspended by a mesorchium.


Development ◽  
1974 ◽  
Vol 32 (1) ◽  
pp. 195-215
Author(s):  
Noriyuki Satoh

Electron-microscopic observations on sex differentiation during normal development in the medaka, Oryzias latipes, are presented in this report. Primordial germ cells in embryos 6–10 days after fertilization are clearly distinguishable from somatic cells by the presence in the cytoplasm of the former of germinal dense bodies, closely associated with large aggregations of mitochondria. The gonadal primordium is composed of the primordial germ cells and the enveloping somatic cells; no special ultrastructural relationships have been detected between these two cell types. The sex of an embryo cannot be decided by means of the electron microscope. The newly formed ovary is distinguishable in newly hatched fry of about 5 mm total body length. In the ovary, a layer of cells forming the ovarian wall encloses the ovarian matrix, consisting of oogonia and their surrounding follicle cells. There are often observable desmosomes between two neighbouring follicle cells. Although marked rearrangement of somatic cells takes place during sex differentiation of the gonad, few visible changes are observed in the ultrastructure of the primordial germ cells during their transition to oogonia. The transition from the oogonium to the oocyte is, however, characterized by a distinctive change in the nucleus, associated with the onset of meiosis. In the young ovary of 5-day-old fry, ooctye chromatin is visibly organized into electron-dense axial elements at the leptotene stage of meiotic prophase. But in the ovary of 10-day-old fry (about 6 mm body length), many oocytes at zygotene or pachytene stages are found, with synaptonemal complex configurations essentially the same as those described in numerous other meiosing plant and animal cells. In contrast, the testis of newly hatched male fry remains in an undifferentiated state, with the somatic cells simply enveloping the spermatogonia. No cells with the ultrastructural characteristics of Leydig cells (as observed in adult testis) can be distinguished in the young ovary or in the undifferentiated testis 10 days after hatching. Cells with the ultrastructural characteristics of adult Leydig cells are detectable for the first time in the matrix of the testis of 25-day-old young, about 8 mm in total length. When proliferation of spermatogonia has occurred, to form the typical testis of 45-day-old (about 15 mm), young, these cells appear in the interstitial region of the testis. Observations from the present study indicate that sex differentiation of germ cells and somatic cells in the gonad precedes the differentiation of steroid-secreting cells. Therefore, the hypothesis that sex hormones are natural sex-inducers is not supported by the present results. The possibility is emphasized that some intracellular mechanisms may be involved in the natural course of sex differentiation of the germ cells and the gonad in this fish.


Endocrinology ◽  
2007 ◽  
Vol 149 (4) ◽  
pp. 1813-1819 ◽  
Author(s):  
Eri Shiraishi ◽  
Norifumi Yoshinaga ◽  
Takeshi Miura ◽  
Hayato Yokoi ◽  
Yuko Wakamatsu ◽  
...  

Müllerian inhibiting substance (MIS) is a glycoprotein belonging to the TGF-β superfamily. In mammals, MIS is responsible for the regression of Müllerian ducts in the male fetus. However, the role of MIS in gonadal sex differentiation of teleost fish, which have no Müllerian ducts, has yet to be clarified. In the present study, we examined the expression pattern of mis and mis type 2 receptor (misr2) mRNAs and the function of MIS signaling in early gonadal differentiation in medaka (teleost, Oryzias latipes). In situ hybridization showed that both mis and misr2 mRNAs were expressed in the somatic cells surrounding the germ cells of both sexes during early sex differentiation. Loss-of-function of either MIS or MIS type II receptor (MISRII) in medaka resulted in suppression of germ cell proliferation during sex differentiation. These results were supported by cell proliferation assay using 5-bromo-2′-deoxyuridine labeling analysis. Treatment of tissue fragments containing germ cells with recombinant eel MIS significantly induced germ cell proliferation in both sexes compared with the untreated control. On the other hand, culture of tissue fragments from the MIS- or MISRII-defective embryos inhibited proliferation of germ cells in both sexes. Moreover, treatment with recombinant eel MIS in the MIS-defective embryos dose-dependently increased germ cell number in both sexes, whereas in the MISRII-defective embryos, it did not permit proliferation of germ cells. These results suggest that in medaka, MIS indirectly stimulates germ cell proliferation through MISRII, expressed in the somatic cells immediately after they reach the gonadal primordium.


Development ◽  
1976 ◽  
Vol 35 (3) ◽  
pp. 637-648
Author(s):  
Par Eliane Didier ◽  
Noël Fargeix

Quantitative aspects of the colonization of the gonads by germ cells in the quail embryo (Coturnix coturnix japonica) A quantitative analysis made on quail embryos coming from 13 isolated parent couples reveals some significative variations of a genetic origin, between some of the off spring studied: the differences observed concern both the quantitative importance of the colonization of gonads by germ cells and the asymmetrical distribution of PGCs (primordial germ cells) between the two genital ridges. The chronological study of the colonization in the quail shows, as in both the duck and the chick, two periods of rapid and regular increase of the number of gonadic PGC, at stages from 13 to 18 and from 24 to 30 of Hamburger & Hamilton. The distribution of germ cells between the two genital ridges is, at the beginning of the colonization, not very asymmetrical. Between stages 18 and 24 the asymmetry increases and remains stable so that the mean value of D % (percentage of the number of PGC contained in the right gonad) from that moment on is equal to 29–34 %. This value is specific for the quail embryo.


2005 ◽  
Vol 17 (2) ◽  
pp. 236
Author(s):  
T. Horii ◽  
Y. Nagao ◽  
M. Kimura ◽  
I. Hatada

Mammalian parthenotes cannot develop normally to term. Mouse parthenogenetic embryos die by Day 10 of gestation. On the other hand, viable parthenogenetic chimeras were produced by normal host embryos, although parthenogenetic cells were observed in a limited number of tissues and organs and, even in these instances, their contribution was substantially reduced. This can be explained by the aberrant expressions of imprinted genes in parthenogenetic cells. In female mice, erasure of imprints occurs around the time that primordial germ cells enter the gonad, and establishment of imprints occurs in the postnatal growth phase of oogenesis. In this study, we investigated whether aberrant imprints in parthenogenetic embryonic stem (PgES) cells can be erased through the germline. Diploid parthenogenetic embryos were produced by activation of (CBA × C57BL/6-EGFP) F1 mouse superovulated unfertilized oocytes by exposure to Sr2+ and cytochalasin B. Ten parthenogenetic blastocysts were plated and three PgES cell lines were isolated. Chimeras were made by injecting 10–15 PgES cells into ICR(CD-1) mouse blastocysts. Chimeras and chimeric tissues were detected by fluorescent microscopy. In all, 173 chimeric blastocysts were transferred to 9 recipient females, and 101 live pups containing 9 female and 21 male chimeras were born. No significant growth retardation was apparent in PgES chimeras, irrespective of their degree of chimerism. In 5 male chimeras killed at 1 day postpartum (dpp), PgES cells showed a restricted tissue contribution. The contribution to lung, liver, and intestine was considerably lower than in the other tissues such as brain, heart, spleen, and kidney. PgES derived or host embryo derived non-growing oocytes were isolated from dissociated ovaries of female chimeras at 1 dpp under fluorescent microscopy. Methylation imprints in non-growing oocytes were analyzed for maternally methylated imprinted genes Peg1, Snrpn, and Igf2r by the combined bisulfite restriction analysis (COBRA). In normal oocytes, imprints are expected to be erased and these genes are unmethylated at this stage. We observed that these genes were unmethylated in both PgES derived and host embryo derived non-growing oocytes. These results suggest that aberrant imprints in PgES cells can also be erased normally through the germline.


2018 ◽  
Vol 100 (3) ◽  
pp. 697-710 ◽  
Author(s):  
Akihiko Sakashita ◽  
Takuya Wakai ◽  
Yukiko Kawabata ◽  
Chiaki Nishimura ◽  
Yusuke Sotomaru ◽  
...  

Abstract The fertility of sex-reversed XY female mice is severely impaired by a massive loss of oocytes and failure of meiotic progression. This phenomenon remains an outstanding mystery. We sought to determine the molecular etiology of XY oocyte dysfunction by generating sex-reversed females that bear genetic ablation of Sry, a vital sex determination gene, on an inbred C57BL/6 background. These mutant mice, termed XYsry− mutants, showed severe attrition of germ cells during fetal development, resulting in the depletion of ovarian germ cells prior to sexual maturation. Comprehensive transcriptome analyses of primordial germ cells (PGCs) and postnatal oocytes demonstrated that XYsry− females had deviated significantly from normal developmental processes during the stages of mitotic proliferation. The impaired proliferation of XYsry− PGCs was associated with aberrant β-catenin signaling and the excessive expression of transposable elements. Upon entry to the meiotic stage, XYsry− oocytes demonstrated extensive defects, including the impairment of crossover formation, the failure of primordial follicle maintenance, and no capacity for embryo development. Together, these results suggest potential molecular causes for germ cell disruption in sex-reversed female mice, thereby providing insights into disorders of sex differentiation in humans, such as “Swyer syndrome,” in which patients with an XY karyotype present as typical females and are infertile.


Author(s):  
E. M. Eddy

Primordial germ cells are readily recognizable in embryos of the rat due to their large size, generally rounded shape and prominent nuclei with uniformly dispersed heterochromatin. They often have blunted pseudopodal processes at one end and small ruffles or trailing processes at the other, characteristics expected from their known ameboid activity- and migratory abilities. Also, the cytoplasm is rich in polyribosomes and contains a modest amount of rough endoplasmic reticulum and the mitochondria are frequently larger and less dense than those of adjacent somatic cells.In addition to these general characteristics, there are features unique to germ cells which allow them to be identified with certainty. These are: 1) small vesicles containing an irregular, dense core and 2) discrete accumulations of fibrous material known as nuage. Both of these features are present in other species and at other times in the life history of germ cells. The dense-cored vesicles have been noted in fetal and early postnatal mouse oogonia and oocytes, and in hamster and rabbit oocytes.


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