Nuclear ultrastructure in bovine oocytes after inhibition of meiosis by chemical and biological inhibitors

Various components of the ovarian follicle as well as different chemicals can suppress the resumption of meiosis in cumulus-oocyte complexes (COCs). In this study the nuclear ultrastructure of bovine COCs was assessed after 8 h of meiotic inhibition with 50 muM roscovitine (ROSC), 50 muM butyrolactone (BL-I), 2 mM 6-DMAP, 2 muM cycloheximide (CX), or a theca cell monolayer (TC). COCs were recovered according to standard in vitro methods, cultured in a simple and defined medium, and processed for transmission electron microscopy. Control COCs were processed before onset of culture and multiple oocytes were evaluated for each treatment. In all groups, the oocyte nucleus presented a dense fibrillar nucleolus consisting of a fibrillar sphere with a fibrillar center. In TC and 6-DMAP inhibited COCs condensed chromatin adhered to the nucleolus while in all other groups the perinuclear chromatin was separated from the nucleolus. In ROSC inhibited COCs, the nuclear envelope presented only slight small amplitude undulation. The BL-I-inhibited COCs presented an intermediate level of low amplitude undulation of the NE, In CX, 6-DMAP, and TC inhibited COCs the nuclear envelope presented extensively low amplitude undulations. In ROSC inhibited COCs, electron-dense granules formed ring-shaped structures. In some of the BL-I inhibited COCs multiple stellate crystal-like structures were found, and in these COCs the nuclear envelope and the perinuclear cisternae appeared less distinct than in the other BL-I inhibited COCs. In 6-DMAP inhibited COCs interchromatin-like granule clusters were present. In conclusion, the oocyte nuclei in all COCs presented a dense fibrillar nucleolus resembling that in control COCs. However, variations were observed in 1) the nuclear envelope morphology; 2) the chromatin location in relation to the nucleolus; and 3) the presence of different populations of intranuclear granules. Although all treatments inhibited oocyte nucleus breakdown, the mechanisms underlying these effects are different and require further characterization. (C) 2001 Wiley-Liss, Inc.