Penetration in vitro of bovine oocytes during maturation by frozen-thawed spermatozoa.
Penetration in vitro of bovine oocytes during maturation by frozen-thawed spermatozoa.
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冻融精子成熟过程中牛卵母细胞的体外渗透。
DOI：
10.1530/jrf.0.0910329
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发表时间：
1991
期刊：
影响因子：
--
通讯作者：
K. Okuda
中科院分区：
文献类型：
--
作者：
K. Niwa;C. K. Park;K. Okuda
在 TC-199 培养基中培养不同时间的牛未成熟卵母细胞，用含有咖啡因 (5 mM) 和肝素 (10 微克/ml) 的 BO 培养基中的冻融精子进行授精。所有培养0-20小时的卵母细胞均获得了非常高的渗透率（95-100%）。培养0和4 h的卵母细胞授精时，100%穿透并具有解凝精子头，授精后20-22 h，大部分卵母细胞仍处于凝缩生发囊泡(GV)至末期I阶段。首次在培养后 8 小时受精的卵母细胞中观察到雄性和雌性原核的形成。随着卵母细胞成熟的进行，多精比例和穿透卵母细胞的平均精子数逐渐下降。至少一个具有去浓缩头部的精子在GV阶段（未经培养）的卵母细胞的渗透几乎在授精后8小时内完成，并且此时大多数渗透的卵母细胞仍处于GV或浓缩GV阶段。这些结果表明，在所使用的体外条件下，牛卵母细胞的成熟不是精子穿透卵黄或精子核解凝所必需的。
Bovine immature oocytes cultured for various times in TC-199 medium were inseminated with frozen-thawed spermatozoa in Medium BO with caffeine (5 mM) and heparin (10 micrograms/ml). Very high penetration rates (95-100%) were obtained in all oocytes which had been cultured for 0-20 h. When oocytes cultured for 0 and 4 h were inseminated, 100% of them were penetrated and had a decondensing sperm head and most of the oocytes remained at the stage of condensed germinal vesicle (GV) to telophase-I 20-22 h after insemination. The formation of male and female pronuclei was first observed in oocytes inseminated 8 h after culture. The proportions of polyspermy and average number of spermatozoa in penetrated oocytes gradually decreased as oocyte maturation proceeded. Penetration of at least one spermatozoon with a decondensing head into oocytes at the GV stage (without culture) was almost completed up to 8 h after insemination and at that time most of the penetrated oocytes were still at the stage of GV or condensed GV. These results indicate that maturation of bovine oocytes is not required for sperm penetration into the vitellus or for sperm nuclear decondensation under the in-vitro conditions used.