Otoi T, Yamamoto K, Koyama N, Tachikawa S, Suzuki T
Tokushima Prefectural Beef Cattle and Swine Experiment Station Anan, Tokushima 774 Japan.
Theriogenology. 1997 Oct 1;48(5):769-74. doi: 10.1016/s0093-691x(97)00300-2.
This study was conducted to determine the diameter of bovine oocytes that were able to attain their full developmental competence to blastocysts. Oocytes were recovered by aspiration of surface-visible follicles (1 to 7 mm in diameter) from slaughterhouse ovaries. Only healthy-looking cumulus-oocyte complexes were used for in vitro maturation, and they were divided into six groups based on diameter: < 110 microm, 110 to < 115 microm, 115 to < 120 microm, 120 to < 125 microm, 125 to < 130 microm and >/= 130 microm. Oocytes were processed through standard procedures for in vitro maturation, fertilization and culture. Following in vitro maturation or fertilization, some oocytes were stained to assess nuclear maturation and penetration rates. The numbers of embryos that cleaved at 42 h post insemination and developed to blastocysts and hatched blastocysts after 8 days of culture were recorded. The mean oocyte diameters were 114.0 +/- 4.8 microm. The oocytes displayed size-related ability to undergo meiotic maturation. The rates of nuclear maturation of oocytes in the greater than 115-microm size range were significantly higher than those of oocytes with diameters < 115 microm. In the < 120 microm diameter groups, the polyspermic fertilization rates of oocytes < 115 microm were significantly higher than those of oocytes 115 to < 120 microm in diameter. The rates of cleavage and development to blastocysts and hatched blastocysts rose as oocyte diameter increased. Among oocytes with a diameter >110 microm, oocytes < 120 microm were found to have significantly lower developmental competence than oocytes 120 to < 130 microm in diameter. These results suggest that bovine oocytes have acquired full meiotic competence at a diameter of 115 microm but not yet attained full developmental competence to blastocysts, and that oocytes have acquired full developmental competence at a diameter of 120 microm.
本研究旨在确定能够发育至囊胚并具备完全发育能力的牛卵母细胞的直径。通过抽吸屠宰场卵巢表面可见的卵泡(直径1至7毫米)来回收卵母细胞。仅使用外观健康的卵丘 - 卵母细胞复合体进行体外成熟,并根据直径将它们分为六组:<110微米、110至<115微米、115至<120微米、120至<125微米、125至<130微米和≥130微米。卵母细胞经过体外成熟、受精和培养的标准程序处理。在体外成熟或受精后,对部分卵母细胞进行染色以评估核成熟和穿透率。记录授精后42小时分裂的胚胎数量以及培养8天后发育至囊胚和孵化囊胚的数量。卵母细胞的平均直径为114.0±4.8微米。卵母细胞表现出与大小相关的减数分裂成熟能力。直径大于115微米的卵母细胞核成熟率显著高于直径<115微米的卵母细胞。在直径<120微米的组中,直径<115微米的卵母细胞多精受精率显著高于直径115至<120微米的卵母细胞。随着卵母细胞直径的增加,分裂率以及发育至囊胚和孵化囊胚的比率上升。在直径>110微米的卵母细胞中,发现直径<120微米的卵母细胞发育能力显著低于直径120至<130微米的卵母细胞。这些结果表明,牛卵母细胞在直径为115微米时已获得完全减数分裂能力,但尚未获得发育至囊胚的完全发育能力,而卵母细胞在直径为120微米时已获得完全发育能力。