Chian R C, Sirard M A
Départemente des sciences animales, Université Laval Québec, Canada.
Mol Reprod Dev. 1995 Dec;42(4):425-31. doi: 10.1002/mrd.1080420408.
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 microgram/ml follicle-stimulating hormone (FSH), 50 micrograms/ml luteinizing hormone (LH), and 1.0 microgram/ml estradiol-1 beta (E2). When oocytes with cumulus cells were cultured in different media, MM along, MM + 0.5 microgram/ml FSH, MM + 50 micrograms/ml LH, MM + 1.0 microgram/ml E2, 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 labelled with [35S]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 micrograms/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.
评估了在不同成熟条件下培养30小时后的牛卵母细胞对孤雌激活（钙离子载体A23187）的反应。当带有或不带有卵丘细胞的卵母细胞单独在含10%胎牛血清的TCM - 199（成熟培养基[MM]）中培养时，对10μM A23187反应的卵母细胞激活率相似（分别为86%±4和89%±8）。然而，当MM中添加0.5μg/ml促卵泡激素（FSH）、50μg/ml促黄体生成素（LH）和1.0μg/mlβ - 雌二醇（E2）时，带有卵丘细胞成熟的卵母细胞激活率显著低于不带有卵丘细胞的卵母细胞（分别为48±10和79%±16；P<0.05）。当带有卵丘细胞的卵母细胞在不同培养基中培养时，即单独的MM、添加0.5μg/ml FSH的MM、添加50μg/ml LH的MM、添加1.0μg/ml E2的MM以及添加所有三种激素的MM，激活的卵母细胞百分比分别为91%±3、52%±14、83%±7、83%±13和54%±7。培养过程中带有或不带有卵丘细胞的卵母细胞在激活后5小时开始形成原核（分别为5%±1和7%±）。最后，用[³⁵S]甲硫氨酸标记激活前（3小时）和激活后（10小时）带有或不带有卵丘细胞成熟的卵母细胞合成的蛋白质进行分析。激活前，带有完整卵丘细胞的卵母细胞产生一种47kD的蛋白质复合物。在含0.5μg/ml FSH的MM中培养的带有卵丘细胞的卵母细胞，其特征是有60kD的活跃蛋白质合成。激活后，在成熟期间，带有卵丘细胞的卵母细胞中出现了75kD和47kD的蛋白质，而不带有卵丘细胞的卵母细胞中则没有。这些结果表明，体外成熟的牛卵母细胞对孤雌激活的能力因卵丘细胞的有无以及培养期间MM中FSH的存在与否而有所不同。
