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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 190 (1994), S. 263-271 
    ISSN: 1432-0568
    Keywords: Infrequent structures ; Oocytes ; Cattle ; Zebu
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Uncommon and controversial structures in cattle oocytes were studied in dominant and subordinate follicles of unstimulated Bos taurus and zebu (Bos indicus), and in FSH-stimulated B. taurus cattle, before or after administration of cloprostenol. Growing oocytes were very rare in follicles more than or equal to 5 mm in diameter. For the first time, special vesicles that may be involved in cortical granule synthesis were observed. Classical rough endoplasmic reticulum was very rare in unstimulated oocytes, but was seen at superovulation. Vacuolation of the nucleolus was a common feature in dominant oocytes that were collected after cloprostenol injection, except in superovulated cattle. Annulate lamellae were very rare or absent in oocytes of B. taurus, while they were more frequent in zebu oocytes.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Anatomy and embryology 190 (1994), S. 583-590 
    ISSN: 1432-0568
    Keywords: Cattle ; Epithelium ; Oviduct ; Secretion ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The non-ciliated (NC) cells of the bovine oviduct epithelium, have been shown to release embryotrophic substances to the oviduct lumen. The aim of the present study was to investigate the ultrastructure, focusing on aspects of the secretory machinery, of NC cells in different segments of the oviduct during and after transoviduct migration of zygotes and embryos. Dairy heifers (n=8) were superovulated with an ECG/cloprostenol regimen, and the time of ovulation was estimated by ultrasound scanning. Samples from the infundibulum, ampulla, isthmus and uterotubal-junction of the oviduct were surgically collected from animals at 19–96 h and 7 1/2; –8 1/2 days after ovulation and processed for transmission electron microscopy, following standard procedures. The NC cells contained characteristic membrane-bound secretory granules composed of a lamellar cortex encaging an amorphous medulla. The two components could still be recognized during extrusion of the granule content into the oviduct lumen by exocytosis. During granulogenesis, small maturing granules without the lamellar structure were observed, but distinct condensing vacuoles were absent. An abundance of granules was found in the early versus the late group. In both groups the uterotubual junction was almost free of granules. This segment, on the contrary, was characterized by the presence of primary and secondary lysosome-like bodies. In the early group the intracellular location of the granules varied between oviduct segments. In the infundibulum they were placed in the supranuclear cytoplasm, in the isthmus they were found in the most apical part of the cells, while in the ampulla an intermediate granule position was noticed. In both groups the uterotubal junction was almost free of granules. This segment, on the contrary, was characterized by the presence of primary and secondary lysosome-like bodies. Cytoplasmic protrusions, often containing nuclei, were more frequent in the late than in the early group. This phenomenon may represent epithelial renewal. In conclusion, the NC cell of the bovine oviduct epithelium possesses an extensive capacity for protein synthesis and secretion. The numbers and location of secretory granules show cyclic and segmental variations. Most granules are present in the infundibulum and ampulla during the period of transoviduct migration of zygote and embryo.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Anatomy and embryology 179 (1989), S. 435-442 
    ISSN: 1432-0568
    Keywords: Ultrastructure ; In vitro fertilization ; Bovine ; Ova ; Cortical granules
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Heifers were superovulated by PMSG or FSH, and oestrus was induced by prostaglandin. One group of animals was ovariectomized 19–26 h after the LH peak, the content of preovulatory follicles aspirated, and the oocytes processed for in vitro fertilization. Another group was inseminated and ova were collected from the oviducts for study of in vivo fertilization. All ova were examined ultrastructurally. The developmental rate following in vitro fertilization was delayed compared to fertilization in vivo. A high proportion of the in vitro fertilized ova showed polyspermic penetration of the zona pellucida, and supernumerary spermatozoa were found in the ooplasm of some ova. In vivo fertilization was associated with release and subsequent dispersal of the cortical granule content in the perivitelline space. In contrast to this the released granule content of the in vitro fertilized ova remained undispersed close to the oolemma. This feature may account for the high incidence of polyspermic penetration of the zona pellucida. In addition, the study provided an ultrastructural visualization of the initial contact between the equatorial segment of the spermatozoon and the microvilli of the oocyte, and the subsequent internalization of the sperm head.
    Type of Medium: Electronic Resource
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  • 6
    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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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Anatomy and embryology 176 (1987), S. 41-44 
    ISSN: 1432-0568
    Keywords: Bovine ; Oocyte maturation ; Meiosis ; Gap junction ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Cumulus-oocyte complexes were obtained from cows by aspiration of small (1–6 mm in diameter) antral follicles after slaughter. Complexes with a compact multilayered cumulus investment were cultured and processed for transmission electron microscopy after different periods of culture including a 0 h control group. In 0 h control oocytes the cumulus cells had numerous projections which penetrated the zona pellucida and established gap junctions with the oolemma. A partial loss of these junctions was noticed as an early event of oocyte maturation occurring within the first 3 h of culture. A low frequency of gap junctions was maintained until 12–18 h of culture where the junctional contact was completely disrupted. This decrease in intercellular communication was parallelled by resumption of oocyte meiosis.
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Anatomy and embryology 190 (1994), S. 65-72 
    ISSN: 1432-0568
    Keywords: Bovine ; Cumulus ; Corona ; Oocyte ; Expansion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Bovine cumulus-oocyte complexes with or without attached pieces of the mural stratum granulosum were matured with or without FSH in vitro for 24 h. Some complexes were ooplasmectomized (removal of ooplasm) at the onset of culture or after 9 h of culture, and in others the complete oocyte, including the zona pellucida, was removed. At the end of culture, the degree and morphology of corona and cumulus expansion were determined by a subjective scoring system, transmission electron microscopy followed by computerized image analysis, or scanning electron microscopy. In the absence of FSH no expansion was seen. In the presence of FSH, hyaluronidase-sensitive cumulus expansion was observed in intact complexes and complexes ooplasmectomized after 9 h of culture. In complexes ooplasmectomized at the start of culture, hyaluronidase-insensitive expansion of the corona compartment was seen as well. A similar reaction was found in free corona and cumulus cell masses cultured in the presence or absence of oocytes. Mural granulosa in intact and ooplasmectomized complexes expanded only slightly under the influence of FSH and exhibited no hyaluronidase sensitivity. Ooplasmectomy at the start of culture increased the frequency of ruffled cell membranes and decreased the occurrence of intercellular bridges as compared with unmanipulated complexes cultured under corresponding conditions. It is concluded that FSH-induced cumulus expansion and hyaluronidase-sensitive extracellular mucus production of bovine cumulus investment in vitro are not dependent on the oocyte, except for minor changes in the surface morphology of the cumulus cells.
    Type of Medium: Electronic Resource
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Anatomy and embryology 195 (1997), S. 327-336 
    ISSN: 1432-0568
    Keywords: Key words Cattle ; Ooplasm ; Organelles ; Growth
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  The aim of the present study was to describe in detail the changes occurring in the cytoplasmic ultrastructure of the bovine oocyte from the onset of growth in the primordial follicle until the completion of growth in the tertiary follicle. Bovine oocytes from primordial, primary, secondary and early to mid-antral follicles were processed and analysed by light and transmission electron microscopy. The primordial follicular oocyte was characterized by numerous coated pits on the oolemma and the accumulation of free and organelle-related smooth (SER) and rough (RER) endoplasmic reticulum, round mitochondria and Golgi complexes around the nucleus, which was located slightly off centre. Up to the secondary follicular stage the oocyte displayed an increase in the number of microvilli, elongated mitochondria and Golgi complexes. During the secondary follicular stage, formation of the zona pellucida, development of gap junctions between the oocyte and the granulosa cells, formation of the cortical granules in the oocyte and reduction in the number of coated pits on the oolemma were seen. In the tertiary follicular oocyte up to 100 μm in diameter, the number of Golgi complexes and lipid droplets increased and the organelles were dislocated to the deep cortical region. During the final growth of the oocyte up to 〉120 μm, the organelles were dislocated further to the peripheral region, the extent of the free SER and RER compartments were reduced, the number of individual cortical granules increased, hooded mitochondria became abundant and the perivitelline space developed. In conclusion, the growth of the bovine oocyte is associated with the relocation and modulation of a number of cytoplasmic organelles as well as the development of oocyte specific structures such as the zona pellucida and cortical granules.
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  • 10
    Electronic Resource
    Electronic Resource
    Springer
    Anatomy and embryology 181 (1990), S. 325-331 
    ISSN: 1432-0568
    Keywords: Oocyte maturation ; Ultrastructure ; Fox ; Ovulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Blood samples were taken weekly from seventeen mature blue fox vixens (average age five years), from late anoestrus until pro-oestrus, and then taken daily. The vixens were sacrificed at various stages of oestrus, and oocytes were collected from ovarian follicles by aspiration, and/or from oviducts by flushing. The structural features of oocyte maturation were related to the time of the luteinizing hormone (LH) peak. On days 1–2 after the LH peak the oocyte nucleus migrated from a central to a peripheral position in the ooplasm and assumed a flattened appearance. The cumulus investment expanded simultaneously and ovulation took place around day 2. On days 2–3 the oocyte nuclear envelope broke down, the nucleoli disappeared, the metaphase of the first meiotic division was reached, the Golgi complexes decreased in size, the perivitelline space enlarged, and all junctional contact between cumulus cell projections and oocyte was disrupted. On days 3–5 the first polar body was extruded, the metaphase of the second meiotic division was reached, and the cumulus cells degenerated. On day 5 the release of cortical granule content was occasionally seen, and from day 6 the oocytes showed signs of degeneration. In a few animals deviant oocyte maturation was noticed.
    Type of Medium: Electronic Resource
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