Bovine cumulus-oocyte-complex-quality is reflected in sensitivity for alpha-amanitin, oocyte-diameter and developmental capacity.
The aim of the present study was to find more parameters to define developmental competence of cumulus-oocyte-complexes (COCs). Bovine COCs were divided into five groups based on their morphology. In order of increasing level of atresia: COC-A had a bright, compact cumulus investment; COC-B1 also had a compact cumulus investment, but was darker than COC-A; the color of COC-B2 was comparable with COC-B1 but the corona radiata appeared to dissociate from the rest of the cumulus investment; the cumulus of COC-B3 was almost black and the corona radiata was almost completely dissociated from the rest of the cumulus investment; COC-C had a strongly expanded cumulus investment with dark spots of degenerated cells. An increasing level of atresia was accompanied by: (1) an increasing zona pellucida diameter (147.6, 150.8, 151.0, 154.3 and 155.1 microm, respectively, for COC-A, COC-B1, COC-B2, COC-B3 and COC-C); (2) an increasing oocyte diameter except for COC-C (120.9, 122.8, 122.8, 123.9 and 118.4 microm, respectively); (3) an increasing developmental competence except for COC-C (13.9, 14.7, 17.4, 19.1 and 11.5%, respectively, development to morula and blastocyst after in vitro embryo production (IVP)) and (4) by a increasing percentage of oocytes exhibiting germinal vesicle breakdown (GVBD) after 24h culture with alpha-amanitin and FSH, except for COC-A (34.4, 68.6, 51.0, 22.0 and 4.8%, respectively, oocytes arrested in GV). In general, embryo-quality, expressed in nuclei-count, was significantly affected by COC-quality (P < 0.05) with B3 > B1> B2 > A > C. However, if the developmental stages were compared separately, this effect was less evident. It was found that, in absence of FSH, alpha-amanitin was unable to inhibit GVBD and that the success of GVBD inhibition was positively correlated to the amount of cumulus cells surrounding the oocyte. COCs that had been exposed to alpha-amanitin and FSH during maturation retained the ability to cleave after IVF, but were unable to develop any further.
de Wit
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Institute for Animal Science and Health (ID-DLO), P.O. Box 65, 8200 AB, Lelystad, The Netherlands. a.a.c.dewit@id.dlo.nl
Kruip
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D000546:Amanitins / Q000494:pharmacology*; D000818:Animals; D002417:Cattle / Q000502:physiology*; D004396:Coloring Agents; D005314:Embryonic and Fetal Development / Q000187:drug effects / Q000502:physiology*; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary; D005640:Follicle Stimulating Hormone / Q000494:pharmacology; D008297:Male; D008858:Microscopy, Phase-Contrast / Q000662:veterinary; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology*; D009865:Oocytes / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D010078:Oxazines; D012044:Regression Analysis; D013194:Staining and Labeling; D015044:Zona Pellucida / Q000187:drug effects / Q000502:physiology
D000546:Amanitins; D004396:Coloring Agents; D019384:Nucleic Acid Synthesis Inhibitors; D010078:Oxazines; C018437:aceto-orcein; D005640:Follicle Stimulating Hormone