Kim N H, Funahashi H, Abeydeera L R, Moon S J, Prather R S, Day B N
Department of Animal Sciences, University of Missouri-Columbia 65211, USA.
J Reprod Fertil. 1996 May;107(1):79-86. doi: 10.1530/jrf.0.1070079.
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.
研究了输卵管液对猪精子穿透和皮质颗粒胞吐作用的影响。通过荧光标记凝集素染色和激光扫描共聚焦显微镜观察体内和体外成熟卵母细胞的皮质颗粒胞吐作用。体外受精后，成熟卵母细胞的胞吐作用分为三类：皮质颗粒完全胞吐且渗出物均匀分布于整个卵周隙（I型）；完全胞吐但渗出物部分分布（II型）和皮质颗粒不完全胞吐（III型）。I型胞吐作用的卵母细胞发生率在体内成熟的卵母细胞中高于体外成熟的卵母细胞。向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以及穿透卵中存在的精子平均数量。精子穿透后，在存在1%输卵管液的情况下，皮质颗粒渗出物的分布与对照组无差异。在授精前将卵母细胞在含有10%或30%输卵管液的培养基中培养1.5小时，单精受精发生率增加且精子穿透率未降低。在含有30%输卵管液的培养基中对卵母细胞进行预培养可增加I型皮质颗粒反应，并在精子穿透时增加透明带对0.1%（w/v）链霉蛋白酶溶解的抵抗力。这些结果表明，输卵管分泌物中的一种或多种因子对于完全皮质颗粒反应和透明带的修饰是必需的。
