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Development of bovine nuclear transfer embryos made with oogonia.

The pluripotency of embryonic germ cells in the mouse suggests that mitotic bovine fetal germ cells might also be a source of pluripotent cells. To investigate the pluripotency of bovine oogonia, the development in vitro of bovine embryos reconstructed by fusing oogonia with enucleated oocytes was compared with that of embryos made similarly with either blastomeres or granulosa cells. The donor cells (fresh oogonia, cryopreserved oogonia, 16- to 32-cell-stage blastomeres, or granulosa cells) were fused to the enucleated oocytes electrically. The proportions of reconstructed embryos that had cleaved at 40 h after fusion using these types of donor cells were not significantly different (37%, 33%, 56%, and 31%, respectively; p > 0.05). However, the proportions of cleaved reconstructed embryos that developed to the blastocyst stage were 9%, 13%, 36%, and 3%, respectively, significantly higher (p < or = 0.05) with blastomeres than with the other three types of donor cells. After transfer of 3 morulae and 4 blastocysts made with oogonia into three recipient heifers, embryonic and extra-embryonic tissues developed in one animal. On recovery after 43 days gestation, this conceptus was shown to be genetically identical, at 11 microsatellite loci, to the fetus that had provided the oogonia. Cytological analysis of the embryos made with oogonia at 40-44 h after fusion and at the morula and blastocyst stages revealed that aberrant cytokinesis and nucleokinesis had given rise to multinucleated, anucleate, and polyploid cells in the reconstructed embryos. It is concluded that limited pluripotency of bovine oogonia has been demonstrated, warranting further study in this area.

Animals↗

Fine structure of human oogonia in the foetal ovary.

Foetal ovarian tissue is now being cultured or frozen, to generate oocytes for assisted reproduction, an emerging technology. This study examines the ultrastructure of oogonia at 13-15 weeks of gestation, which could be used as a control for culture and freezing of foetal ovaries. Oogonia are largely located in the ovarian cortex, whilst primordial germ cells (PGC) and somatic follicle cells compose the surface epithelium. Oogonia and PGC have large vesicular nuclei with clear cytoplasm, compared to dense follicle cells, which have polymorphic nuclei. Follicle cells intermingle with oogonia and establish close contacts - beginning of folliculogenesis. Nuclei of oogonia contain one to three highly reticulated nucleoli, reflecting high levels of RNA synthesis at the onset of growth. Rough endoplasmic reticulum (RER) form stacks of cisternae associated with numerous ribosomes. Prominent organelles in the ooplasm are elongated mitochondria with dense matrices and tubular cristate presenting a multilocular appearance. Typical Golgi complexes, dense bodies and clear vacuoles are present and microfilaments are located beneath the plasma membrane. The most remarkable feature of oogonia is that they have typical juxtanuclear centrioles (diplosomes) with dense pericentriolar material, which nucleate microtubules, characteristic of functional centrosomes organizing the cytoskeleton. The mature oocyte has no centrioles, since the maternal centrosome is inactivated or reduced, while the paternal is dominant. Centrioles are most likely involved in mitosis of oogonia.

Centrioles↗

Description of primordial germ cells, oogonia, oocytes and embryo-like growth in squash preparations of tissues from hematological malignancies.

This study evidences the presence of primordial germ cells, in tissue squash preparations and sections from hematological malignancies. Primordial germ cells were identified by their morphology, the intense PAS, PAS-D reaction and presence of calcium-activated neutral proteinase. Primordial germ cells gave rise to nuclear vlimata. Immature oogonia exhibited a nuclear envelope and a star-shaped nuclear core, arising from acellular globose bodies impregnated by a nuclear vlima of primordial germ cell. Bone marrow tissue oogonia were PAS and PAS-D positive, identical to fungal ones. Calcium-activated neutral proteinase was demonstrated in the plasma of the acellular globose bodies, the nuclear envelope and the conglomerated primordial germ cells. Immature bone marrow oogonia progressed into mature ones, leptotene, diplotene, dictyotene and mature oocytes. Nuclear vlimata fertilized primordial germ cells, oogonia and oocytes, giving rise to round embryos at the morula and hatching morula-like stages. Embryos consisted of a zonapellucida-like cortex, composed of glycosaminoglycans, glycoproteins, protease and diffuse nuclear material, enclosing developing cells. Primordial germ cells, oogonia and embryos were also demonstrated in squash preparations of adult rat testis and sections of normal rat bone marrow tissues. The observations document that primordial germ cells are the primary stem cells which give rise to nuclear vlimata and oogonia, which constitute the secondary stem germ cells. The results are discussed in terms of stem cell renewal according to the events: primordial germ cells-gametes-fertilization-embryos-primordial germ cells.

Animals↗

[Oogenesis of Tilapia mossambica. I. Oogonia and meiotic prophase oocytes].

Using light and electron microscopy and autoradiography, the morphology and synthesis of DNA, RNA and proteins in oogonia and early meiotic prophase oocytes in Tilaria mossabique were studied. According to dimensions and morphological features observed it is possible to distinguish between two groups of oogonia: large oogonia corresponding to type A spermatogonia of mammals, and small actively dividing oogonia, located in groups and identical to type B spermatogonia. The morphology of oogonia and of the early meiotic prophase oocytes well compares with the pattern described for other species of bony fishes. In the cytoplasm of these cells dense bodies, nuage-material, free ribosomes, large mitochondria with lamellar cristae and Golgi cisterns are available. In the oocyte nuclei at zygotene and pahytene stages 3H-thymidine incorporation was seen mainly into the nucleolus-associated chromatin. Besides, the formation of a heterochromatin cape and the synaptonemal complex was observed. Incorporation of 3H-uridine and 3H-leucine in the nuclei of these cells was very poor.

Animals↗

Immunocytochemical evidence for methylation of the inactive X chromosome in human fetal oogonia.

The state of DNA methylation of the X chromosomes of human interphase oogonia from a 46,XX and a 46,XX/47,XXX fetus at 17 weeks of gestation was tested immunocytochemically with an antibody to 5-methylcytosine (5MeC). Of 1637 oogonial nuclei from the 46,XX fetal ovary, 313 (19.1%) contained Barr bodies, of which 93.6% were positive for 5MeC. Of 1780 oogonia from the 46,XX/47,XXX fetus 327 (18.4%) contained Barr bodies; 175 oogonia had one Barr body and 152 had two. Of the single Barr bodies 145 (82.8%) had positive 5MeC reaction product. Of the 152 oogonia from the XXX line, 97 (63.8%) had positive 5MeC on both Barr bodies, 35 (23%) had one positive and one negative, and 20 (13.1%) had no product on either Barr body. This immunocytochemical evidence supports the hypothesis that the DNA of the inactive X-chromosome of the human 17-week gestation oogonium is methylated.

DNA↗

From oogonia to mature oocytes: inactivation of the maternal centrosome in humans.

The fine structure of human oogonia and growing oocytes has been reviewed in fetal and adult ovaries. Preovulatory maturation and the ultrastructure of stimulated oocytes from the germinal vesicle (GV) stage to metaphase II (MII) stage are also documented. Oogonia have large nuclei, scanty cytoplasm with complex mitochondria. During folliculogenesis, follicle cell processes establish desmosomes and deep gap junctions at the surface of growing oocytes, which are retracted during the final stages of maturation. The zona pellucida is secreted in secondary follicles. Growing oocytes have mitochondria, Golgi, rough endoplasmic reticulum (RER), ribosomes, lysosomes, and lipofuscin bodies, often associated with Balbiani bodies and have nuclei with reticulated nucleoli. Oocytes from antral follicles show numerous surface microvilli and cortical granules (CGs) separated from the oolemma by a band of microfilaments. The CGs are evidently secreted by Golgi membranes. The GV oocytes have peripheral Golgi complexes associated with a single layer of CGs close to the oolemma. They have many lysosomes, and nuclei with dense compact nucleoli. GV breakdown occurs by disorganization of the nuclear envelope and the oocyte enters a transient metaphase I followed by MII, when it is arrested and ovulated. Maturation of oocytes in vitro follows the same pattern of meiosis seen in preovulatory oocytes. The general organization of the human oocyte conforms to that of most other mammals but has some unique features. The MII oocyte has the basic cellular organelles such as mitochondria, smooth endoplasmic reticulum, microfilaments, and microtubules, while Golgi, RER, lysosomes, multivesicular, residual and lipofuscin bodies are very rare. It neither has yolk nor lipid inclusions. Its surface has few microvilli, and 1-3 layers of CGs, aligned beneath the oolemma. Special reference has been made to the reduction and inactivation of the maternal centrosome during oogenesis. The MII spindle, often oriented perpendicular to the oocyte surface, is barrel-shaped, anastral and lacks centrioles. Osmiophilic centrosomes are not demonstrable in human eggs, since the maternal centrosome is nonfunctional. However, oogonia and growing oocytes have typical centrioles, similar to those of somatic cells. The sperm centrosome activates the egg and organizes the sperm aster and mitotic spindles of the embryo, after fertilization.

Centrosome↗

Kinetics of gametogenesis. II. Comparative autoradiographic studies of oogonia and multiplying prospermatogonia of the Wistar rat.

In the rat the last generation of oogonia and multiplying prospermatogonia (M-prospermatogonia), frequently arranged in synchronized clusters, enters mitosis on about day 17 post conception (p.c.). The duration of the S-phase D-S-Duration and the minimal generation time Tmin of both kinds of "gonia" were determined by the method of labeled mitoses (22 female and 22 male fetuses derived from 11 pregnant rats were sacrificed from 2 to 22 h after a single i.p. injection of 3H-thymidine on day 17 p.c.). Three curves, derived from the labeled prophases, metaphases and the postmitotic descendents of oogonia and M-prospermatogonia--oocytes and primary transitional prospermatogonia (T1-prospermatogonia)--were evaluated. It was demonstrated that the curves as well as the calculated values of D-S-Duration and Tmin are very similar for oogonia and M-prospermatogonia. D-S-Duration ranged from about 10 to 12.5h (10 h read off from the curves of labeled metaphases), Tmin from 16.5 to 18 h (16.5 h read off from the curves of labeled metaphases).

Animals↗

Comparative studies of normal and neoplastic ovarian germ cells: 1. Ultrastructure of oogonia and intercellular bridges in the fetal ovary.

Electron microscopic studies were performed on human fetal ovaries to gain insight into the process of mitotic proliferation in developing germ cells. Three stages of germ cell differentiation are present during the early gestational period: primitive germ cells, oogonia, and oocytes. Oogonia, representing the mitotic stage of differentiation, are the predominant cell type present between 9 and 12 weeks' gestation and then progressively decrease in number as a result of transformation into oocytes in meiosis and degeneration. Mitotic division of oogonia, which is extensive during the late first and early second trimesters, is characterized by incomplete separation and persistence of intercellular bridges between germ cells. Intercellular bridges were found in large numbers from 10 weeks until the time of follicle formation at midgestation. The bridges contained microtubule arrays consistent with remnants of the spindle apparatus. The findings support the role of germ cell bridges in maintaining coordination of proliferative activity during the early developmental period.

Cell Differentiation↗

A new species of Pythium with ornamented oogonia: morphology, taxonomy, internal transcribed spacer region of its ribosomal RNA, and its comparison with related species.

Pythium spiculum sp. nov. was isolated from soil samples taken in a vineyard in the Burgundian region of France and from different locations in Spain and Portugal. The oomycete has spiny oogonia and does not sporulate readily. It resembles Pythium mamillatum Meurs, but has its own distinguishing characteristics. It also exhibits sickle-shaped as well as spherical appressoria which at times are associated with sex organs like those found in Pythium abappressorium Paulitz and Pythium contiguanum Paul. Sequencing of the internal transcribed spacer region of its nuclear ribosomal DNA and a close look at its morphological characters have now enabled us to describe it as a new species. The internal transcribed spacer region of its rRNA gene sequence is comprised of 945 bases. This oomycete is closely related to the members that form ornamented or spiny oogonia like Pythiummamillatum, Pythium spinosum and Pythium irregulare but also with those producing smooth-walled oogonia like Pythium paroecandrum, Pythium sylvaticum and Pythium cylindrosporum. Taxonomic description of this new species, its comparison with related oomycetes, the sequence of the internal transcribed spacer region of its rRNA gene and the phylogenetic tree, are given here.

Base Sequence↗

Change in the radiation responses of oogonia in the embryos and fry of the fish Oryzias latipes.

Embryos of the fish Oryzias latipes were irradiated with 1000 R of X-rays 1 day before hatching,and the post-irradiation change in the female germ-cell population was observed. Scarcely any reduction in the number of oogonia was observed, but their proliferation was inhibited. Repopulation occurred between 12 and 20 days after hatching. These responses were quite different from those of germ cells in the irradiated fry (Hamaguchi and Egami 1975). Embryos and/or fry were also exposed to 1000 R of X-rays 1 day before hatching and 0, 1, 2, and 3 days after hatching. A comparison of their responses suggested that the change in the radiation responses of oogonia is correlated with the initiation of meiosis.

Animals↗

[Fetal thymus and thymuline stimulate the in vitro proliferation of oogonia in the fetal rat ovary].

In rat ovaries explanted on day 13.5 p.c. and cultured in vitro for up to 6 days, the number of germ cells is enhanced in thymulin-supplemented medium and/or after co-culture of the ovarian explants with foetal thymic tissue compared to ovaries cultured in synthetic medium. Corticosterone added to the medium prevents the secretion of thymulin by the foetal thymus and in that condition the thymus does not influence the proliferation of oogonia. These results provide additional evidence that the pituitary-adrenal-thymic axis might be involved in the control of oogonia proliferation in vivo, taking into account our previous experimental finding that the number of germ cells is increased in ovaries of hypophysectomized foetuses.

Animals↗

Mutational changes of quantitative morphological traits in the convict cichlid (Cichlasoma nigrofasciatum Guenther) after irradiation of parental spermatogonia and oogonia with different doses of X-rays.

10 continuous and 6 discrete variables of body dimensions as well as 16 proportions calculated from them were determined in postirradiation F1 convict cichlid fish (Cichlasoma nigrofasciatum) which were derived from spermatogonia and oogonia both exposed to either 0 (controls), 250, 500, 750, 1000, or 2000 R of X-rays. While no interactions with sex or radiation dose were found, clear dosage effects and some sex effects occurred. The highest response for almost all quantitative traits examined as well as for variants of coloration or of vertebral aberrations appeared after 250 R to both parental germ cells. The lowest effectiveness was observed for almost all characters under investigation after 0 and 500 R. Accordingly, the largest difference between the 6 different treatment groups was found between 250 and 500 R, followed by 250 and 0, 250 and 1000, and 250 and 750 R to parental gonia. The present finding on the alterations of quantitative morphological traits completely corresponds to the changes of intraspecific aggressiveness as found for the same individual fish in a previous study thus confirming the complete congruence of the mutational response of quantitative morphological characters with that of behavioral traits to ionizing radiation.

Animals↗

Cytological evidence for an inactive X chromosome in murine oogonia.

Purified fetal mouse germ cell preparations have been analyzed for the presence of a heterochromatic X chromosome at cell division. A single large heterochromatic chromosome is found in oogonia stage germ cells, but such a chromosome is not present in 16-day fetal oocytes. These results support the idea that mitotic stage germ cells are subject to X-chromosome inactivation and that they go through a reactivation step at meiosis.

Animals↗

Two progenitor cells for human oogonia inferred from pedigree data and the X-inactivation imprinting model of the fragile-X syndrome.

Laird has proposed that the human fragile-X syndrome is caused by abnormal chromosome imprinting. The analysis presented here supports and extends this proposal. Using published pedigrees that include DNA polymorphism (RFLP) data, we establish that the states of the fragile-X mutation termed "imprinted" and "nonimprinted" usually can be distinguished by the level of cytogenetic expression of the fragile-X chromosome. This information is then used to assess the state of the fragile-X allele in carrier progeny of individual women who inherited a nonimprinted fragile-X chromosome. From this assessment, an estimate is made of the frequency, in individual women, of primary oocytes with an imprinted fragile-X chromosome. The results of this analysis provide additional support for the specific model in which chromosome imprinting occurs in a female in, on average, half of her primary oocytes. This is the expected frequency if X-chromosome inactivation is the initial step in the imprinting of the mutant fragile-X allele. Moreover, this analysis suggests a biological explanation for peculiarities of fragile-X inheritance described by others as "clustering" and the "Sherman paradox." We interpret these peculiarities as consequences of a very small number of oogonial progenitor cells. Two progenitor cells for oogonia is the best integer estimate of the number of such cells at the time of the initial event that leads to chromosome imprinting.

Alleles↗

[Oogonia degeneration in human pre- and perinatal oogenesis].

The number of the degenerating sex cells is extremely small in the period between the 6th and 12th-13th weeks of the intra-uterine development. The cells are represented by isolated oogonia at the interphase or mitosis and by oocytes. The number of the degenerating sex cells increases after the 13th week. Nearly all of them are represented by oocytes at all the stages of the meiosis prophase I. From the 26th week one may observe an intensive degeneration of the cells which is represented mostly by oocytes in diplo- and dictyotene.

Cell Division↗

Pythium ornacarpum: a new species with ornamented oogonia isolated from soil in france.

Pythium ornacarpum sp. nov. was isolated from a soil sample taken from Genlis in the Burgundy region of France. This species is unique because of its ornamented oogonia which are completely surrounded by antheridial filaments. The fungus is closely related to Pythium echinulatum Matthews. Morphological and reproductive aspects of this species as well as a study by PCR of the sequence of the internal transcribed spacer (ITS1) of the nuclear ribosomal gene and its comparison with related species are described here. The nucleotide sequence of the ITS1 region flaking the 5.8S rRNA of this species and other related species are also given here.

Base Sequence↗

Social cohesiveness of convict cichlid fish (Cichlasoma nigrofasciatum Guenther) after irradiation of parental spermatogonia and oogonia with different doses of x-rays.

Comparisons of social cohesiveness were made between F1 convict cichlids (Cichlasoma nigrofasciatum) derived from gonial germ cells which were exposed to 0 (controls), 250, 500, 750, 1000, or 2000 R of X-rays. The cohesiveness was determined by counting the distribution of each ten fish of every F1 group among 12 equal squares within a shallow test tank measuring 80 by 60 by 20 cm. While an increase in cohesiveness was detected in F1 males, as compared to the controls after irradiation with 500 R, the cohesiveness of F1 females decreased after 750 and 2000 R. The increase in male cohesiveness may be associated with a reduction of intermale aggressiveness as found in previous studies.

Aggression↗