title: Stereological characterization of bovine (Bos taurus) cumulus-oocyte complexes aspirated from small antral follicles during the metestrous and proestrous phases.
abstract: Bovine cumulus-oocyte complexes (COCs) aspirated from slaughterhouse ovaries are used for in vitro maturation and fertilization after selection on the basis of morphological appearance of the cumulus and ooplasm. In this context, a quantitative characterization of COCs could provide additional criteria for selecting the most competent complexes. Bovine COCs from small (1-4mm) antral follicles were aspirated from metestrous and proestrous stage ovaries of Holstein-Friesian cows, fixed in glutaraldehyde, randomly embedded in glycol-methacrylate, and sectioned at 20 microm. The unbiased nucleator principle of stereology was used for estimating the mean volumes of complexes, oocytes, cumulus cells, and nuclei of oocytes and cumulus cells. The thickness of the zona pellucida and the relative numerical percentages of the several morphological types (C1, C2 and C3) of cumulus cells were also evaluated. The optical dissector procedure was used for cumulus cell sampling. Quantitative data show that the variability among complexes is generally high, especially for the volume of COCs. There were no linear correlations between the studied parameters, except between the volume of the oocyte and nucleus at metestrus. At proestrus, the volumes of COCs, oocytes and nuclei of oocytes, the volume of follicular cells and the thickness of the inner zona pellucida, were significantly higher than at metestrus. The relative numerical frequency of follicular type C1 cells was lower whereas that of type C3 cells was higher at proestrus than at metestrus. In conclusion, small antral follicles had larger COCs and oocytes at proestrus compared to metestrus and the COCs also had a higher percentage of follicular type C3 cells. Results suggest that for the same type of follicle size there may exist different functional populations of COCs at distinct stages of the bovine ovarian cycle.
authors.affiliation: ICETA-University of Trás-os-Montes and Alto-Douro, 5000-911 Vila Real, Portugal.
authors.forename: A M
authors.initials: AM
authors.lastname: Calado
authors.forename: E
authors.initials: E
authors.lastname: Rocha
authors.forename: A
authors.initials: A
authors.lastname: Colaço
authors.forename: M
authors.initials: M
authors.lastname: Sousa
mesh_terms: D000818:Animals; D002417:Cattle; D002466:Cell Nucleolus / Q000648:ultrastructure; D002467:Cell Nucleus / Q000648:ultrastructure; D048429:Cell Size; D003593:Cytoplasm / Q000648:ultrastructure; D005260:Female; D008686:Metestrus; D009865:Oocytes / Q000648:ultrastructure*; D006080:Ovarian Follicle / Q000648:ultrastructure*; D011359:Proestrus; D013396:Suction; D020858:Tissue and Organ Harvesting / Q000379:methods / Q000662:veterinary; D015044:Zona Pellucida / Q000648:ultrastructure