EFFECTS OF CUMULUS CELLS AND FOLLICLE-STIMULATING-HORMONE DURING IN-VITRO MATURATION ON PARTHENOGENETIC ACTIVATION OF BOVINE OOCYTES

The response to parthenogenetic activation (calcium ionophore A23187) of bovine oocytes after 30 hr of culture in different maturation conditions was evaluated. The activation rates of oocytes in response to 10 mu M A23187 were similar (86% +/- 4 vs. 89% +/- 8) when the oocytes matured with or without cumulus cells were cultured in TCM-199 + 10% fetal calf serum (maturation medium [MM]) alone. However, the activation rates were significantly lower in oocytes matured with than without cumulus cells (48% +/- 10 vs. 79% +/- 16; P < 0.05) when MM was supplemented with 0.5 mu g/ml follicle-stimulating hormone (FSH), 5.0 mu g/ml luteinizing hormone (LH), and 1.0 mu g/ml estradiol-1 beta (E(2)). When oocytes with cumulus cells were cultured in different media, MM alone, MM + 0.5 mu g/ml FSH, MM + 5.0 mu g/ml LH, MM + 1.0 mu g/ml E(2), and MM + all three hormones, the percentages of activated oocytes were 91% +/- 3, 52% +/- 14, 83% +/- 7, 83% +/- 13, and 54% +/- 7, respectively. Oocytes with or without cumulus cells during culture began to form a pronucleus 5 hr after activation (5% +/- 1 and 7% + 1, respectively). Finally, the proteins synthesized by oocytes matured with or without cumulus cells before (3 hr) and after (10 hr) activation was labeled with [S-35]methionine for analysis. Before activation, a 47 kD protein complex was produced by cumulus cell-intact oocytes. An active protein synthesis of 60 kD was characteristic for the oocytes with cumulus cells cultured in MM containing 0.5 mu g/ml FSH. After activation, there was an appearance of proteins at 75 kD and 47 kD in the oocytes with cumulus cells but not in the oocytes without cumulus cells during the maturation period. These results indicate that bovine oocytes matured in vitro have different capacities for parthenogenetic activation depending on the presence or absence of cumulus cells and the presence of FSH in MM during culture. (C) 1995 Wiley-Liss, Inc.