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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Anatomy and embryology 178 (1988), S. 41-46 
    ISSN: 1432-0568
    Keywords: In vitro fertilization ; Bovine ; Ultrastructure ; Ova ; Spermatozoa
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Cumulus-oocyte complexes were collected from cows at an abattoir by aspiration from small (1–6 mm) follicles. The complexes were matured in vitro for 28 h. Subsequently, the cumulus cells and the zona pellucida were removed by enzyme treatment in a proportion of the complexes (zona-free ova). Both cumulus-enclosed and zonafree ova were inseminated in vitro and processed for scanning electron microscopy after different periods of culture. In the cumulus-enclosed ova the number of spermatozoa attached to and penetrating into the cumulus investment increased with increasing culture time. Practically all spermatozoa displayed intact acrosomes. In the zona-free ova clusters of spermatozoa attached to the ovum surface, and at 5 h a proportion of the spermatozoa had undergone the acrosome reaction, and their internalization into the ooplasma was initiated. The acrosome reaction was characterized by an increasing fenestration of the membrane coverings of the acrosomal region of the sperm head. During the sperm head internalization, where the ovum microvilli appeared to contact especially the equatorial segment and the postacrosomal region, the sperm head gradually disappeared from the ovum surface, and the microvilli at the site of internalization became more bulbous. Simultaneous abstriction of the second polar body was seen in some ova.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Anatomy and embryology 178 (1988), S. 47-52 
    ISSN: 1432-0568
    Keywords: Ultrastructure ; In vitro fertilization ; Bovine ; Ova ; Cortical granules
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Cumulus-oocyte complexes collected from cows at an abattoir by aspiration from small (1–6 mm) antral follicles were matured and inseminated in vitro. At different time intervals after insemination the ova were processed for transmission electron microscopy. Up to and including 6 h after insemination all ova were unfertilized, and their cortical granules were more or less clustered. At 6 h acrosome reaction of spermatozoa was observed on the surface of the zona pellucida. At 8 h the first fertilized ovum appeared and the first fully developed spherical pronucleus was observed, at 20 h the first apposition of pronuclei was seen, and at 40 h divisions were ongoing or completed. More than one third of the fertilized ova showed polyspermic penetration of the zona pellucida, and in most of these ova different developmental stages of supernumerary pronucleus formation were observed in the ooplasm. Abnormal cortical granule release was seen in approximately half of the fertilized ova, and it was more frequent in ova with polyspermic as opposed to monospermic penetration of the zona pellucida.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Anatomy and embryology 176 (1987), S. 35-40 
    ISSN: 1432-0568
    Keywords: Bovine ; Oocyte maturation ; Meiosis ; Nucleus ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Cumulus-oocyte complexes were obtained from cow ovaries by aspiration from small (1–6 mm in diameter) antral follicles after slaughter. Complexes with a compact multilayered cumulus investment were cultured and subsequently processed for electron microscopy after various periods of culture. By morphological criteria the oocytes could be divided into the following sequence of meiotic stages. The oocyte nucleus I stage was characterized by a spherical nucleus located peripherally in the ooplasm while undulation of the nuclear envelope and initial chromatin condensation was seen at the oocyte nucleus II stage. The oocyte nucleus breakdown stage was characterized by formation of long slender projections from the nuclear envelope in which the envelope doubled back on itself, appearance of dense areas and haphazardly oriented microtubules in the nucleus, marked condensation of the chromatin, and dissolution of the nuclear envelope into irregular vesicles and tubules. The condensed chromatin I stage was characterized by the location of condensed chromatin configuration and uniformly oriented microtubules in a dense area peripherally in the ooplasm while the final condensed chromatin II stage was characterized by a gradual invasion of condensed chromatin configurations into a dense area combined with the presence of the first polar body in the perivitelline space.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Journal of assisted reproduction and genetics 12 (1995), S. 361-368 
    ISSN: 1573-7330
    Keywords: fetal ova ; in vitro culture ; ultrarapid freezing
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Purpose The present study was performed to culture human fetal ova to determine whether they can be matured and cryopreserved using ultrarapid freezing Methods Thirty-three pairs of fetal ovaries were obtained from fetuses of 16–20 weeks' gestation following elective abortion. Ovarian tissues were minced into approximately 1-mm sizes and cultured in Waymouth media either before or after ultrarapid freezing. The Waymouth medium was supplemented with 15% (v/v) fetal bovine serum, 0.03 IU/ml FSH and 35 ng/ml insulin. The tissue was cultured at 37
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Journal of assisted reproduction and genetics 4 (1987), S. 281-285 
    ISSN: 1573-7330
    Keywords: in vivo maturation ; in vitro maturation ; cattle oocytes ; pregnancy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract In vivo developmental potentials of in vivo and in vitro matured oocytes fertilized in vitro were assessed in cattle. One-cell stages produced from in vivo matured oocytes developed into a pregnancy when transferred to the ampulla part of oviducts of synchronized heifers. In vitro matured oocytes achieved high penetration and cleavage rates but did not develop into pregnancies when transferred to synchronized heifers.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 0148-7280
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Three experiments were performed to evaluate the potential for parthenogenetic activation of bovine oocytes in in vitro fertilization systems and to determine the chromosome complement of the resulting parthenogenotes. In the first experiment, immature oocytes from slaughtered cattles were matured in vitro in Defined Medium (DM) for 24 h to simulate in vitro fertilization conditions. Subsequently, a portion was fixed, and the remainder were transferred to rabbit oviducts. Oocytes were then cultured for 6-8 h or for 24 h with Colcemid present during the last 6 to 8 h and fixed on slides and examined. In the second experiment, mature oocytes were collected from the preovulatory follicles, and the oocytes were subjected to the same culture as in experiment I. In the third experiment, oocytes were treated as in experiment II, except that instead of transfer to rabbit oviducts, they were cultured an additional 48 h in vitro. In experiment I, 131 oocytes were fixed after culture in DM. Of the 79 oocytes analyzed in the pre-rabbit group, 71 (90%) were at the second meiotic metaphase (MII), and 8 (10%) were at pre-MII stage; none were activated. After transfer to rabbits, 291 were fixed. Of these, 80 were analyzed; 37 (46.3%) were MII, 7 (8.6%) were pre-MII, and 36 (45%) were activated. Of the 36 activated oocytes, 26 (72.2%) were haploid, 4 (11.1%) were diploid, 1 (12.8%) was tetraploid, and 5 (13.8%) were in the process of endoreduplication. In experiment II, 51 oocytes were fixed after culture in DM of which 36 (70.6%) could be analyzed; 30 (83.3%) were MII, and 6 (16.7%) were pre-MII. After culture in the rabbit, 68 were fixed of which 27 (39.7%) could be analyzed. Of these 27, 20 (74.1%) were MII, and 7 (25.9%) were activated; 6 were haploid, and 1 was endoreduplicating. In experiment III, 30 oocytes were fixed at the end of the culture period; only 10 could be analyzed of which 8 (80%) were MII and 2 (20%) were pre-MII. In all, 46% of in vitro and 26% of in vivo matured oocytes were activated, based on chromosomal analysis. Of those activated, the majority (74.4%) were haploid, suggesting that activation occurs at or after completion of MII. Endoreduplication appears to be one of the mechanisms leading to the formation of diploid and polyploid parthenogenotes.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
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  • 7
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 31 (1992), S. 249-252 
    ISSN: 1040-452X
    Keywords: Sexual dimorphism ; In vitro ; Early development ; Cattle ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The classical concept of sex determination in mammals is that a Y chromosomal gene controls the development of the indifferent gonad into a testis. Subsequent divergence of sexual phenotypes is secondary to this gonadal determination. The most likely candidate gene is SRY (sex-determining region Y) in humans, and Sry in mouse. However, several lines of evidence indicate that sexual dimorphism occurs even before the indifferent gonad appears. Here we present evidence that bovine male embryos generally develop to more advanced stages than do females during the first 8 days after insemination in vitro. Corresponding relationships between both cell numbers and mitotic indices and sex were also seen. Although it is not clear whether this phenomenon involves factors originating before or after fertilization, these findings suggest that sex-related gene expression affects the development of embryos soon after activation of the embryonic genome and well before gonadal differentiation.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
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