Effects of oviductal fluid on sperm penetration and cortical granule exocytosis during fertilization of pig oocytes in vitro
Effects of oviductal fluid on sperm penetration and cortical granule exocytosis during fertilization of pig oocytes in vitro
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DOI：
10.1530/jrf.0.1070079
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发表时间：
1996-05-01
期刊：
影响因子：
--
通讯作者：
Day, BN
中科院分区：
文献类型：
--
作者：
Kim, NH;Funahashi, H;Day, BN
本文研究了输卵管液对猪精子穿透和皮质颗粒胞吐的影响。用荧光标记凝集素染色和激光共聚焦显微镜观察了体内和体外成熟卵母细胞中皮质颗粒的胞吐作用。成熟卵母细胞在体外受精后的胞吐作用可分为三类：完全的皮质颗粒胞吐作用和渗出液均匀分布在整个卵周间隙（I型）;完全的胞吐作用和渗出液部分分布（II型）和不完全的皮质颗粒胞吐作用（III型）。体内成熟卵母细胞中I型胞吐的发生率高于体外成熟卵母细胞。在受精培养基中添加浓度为1%或10%的输卵管液可降低精子穿透率和穿透卵子中精子的平均数量。精子穿透后，在1%输卵管液中，皮质颗粒渗出物的分布与对照组无差异。受精前卵母细胞在含10%或30%输卵管液的培养基中培养1.5 h，单精受精的发生率增加，而精子穿透率不降低。卵母细胞在含有30%输卵管液的培养基中预培养增加了I型皮质颗粒反应，并在精子穿透时增加了0.1%（w/v）链霉蛋白酶对透明质酸溶解的抵抗力。这些结果表明，输卵管分泌物中的一个因子是完整的皮质颗粒反应和透明细胞修饰所必需的。
The effects of oviductal fluid on sperm penetration and cortical granule exocytosis in pigs were examined. Cortical granule exocytosis in oocytes matured in vivo and in vitro was observed by staining with fluorescent-labelled lectin and laser-scanning confocal microscopy. Exocytosis of matured oocytes was classified into three categories after in vitro fertilization: complete cortical granule exocytosis and even distribution of exudate in the entire perivitelline space (type I); complete exocytosis and partial distribution of exudate (type II) and incomplete cortical granule exocytosis (type III). The incidence of oocytes with type I exocytosis was higher in oocytes matured in vivo than in those matured in vitro. The addition of oviductal fluid at a concentration of 1% or 10% to the fertilization medium decreased sperm penetration and the mean number of spermatozoa present in penetrated eggs. The distribution of cortical granule exudate was not different in the presence of 1% oviductal fluid after sperm penetration from that of control groups. When oocytes were cultured for 1.5 h in medium containing 10% or 30% oviductal fluid before insemination, the incidence of monospermy increased without a decrease in sperm penetration. Preculture of oocytes in medium containing 30% oviductal fluid increased type I cortical granule reaction and increased resistance of the zona pellucida to dissolution by 0.1% (w/v) pronase at the time of sperm penetration. These results suggest that a factor(s) from the oviductal secretion is required for the complete cortical granule reaction and in the modification of the zona pellucida.