Parrish J J, Susko-Parrish J L, Handrow R R, Sims M M, First N L
Department of Meat and Animal Science, University of Wisconsin, Madiso53706.
Biol Reprod. 1989 May;40(5):1020-5. doi: 10.1095/biolreprod40.5.1020.
Oviduct fluid collected from chronically cannulated oviducts of heifers was evaluated for its effect on capacitation of bovine sperm in vitro. Capacitation was determined by the ability of sperm to fertilize bovine oocytes in vitro and to undergo an acrosome reaction (AR) upon exposure to lysophosphatidylcholine (LC). After incubation of sperm with 0-25% (v/v) estrual oviduct fluid (collected +/- 1 day from estrus) for 4 h, addition of LC (100 micrograms/ml) for an additional 0.25 h resulted in an increasing percentage of acrosome-reacted sperm as the concentration of oviduct fluid increased. Sperm incubated 4 h with 25% estrual oviduct fluid fertilized more oocytes than sperm incubated in medium alone (p less than 0.05) but was not different from sperm incubated with 10 micrograms/ml heparin (p greater than 0.05). Glucose inhibited the ability of LC to induce ARs in sperm incubated 4 h with heparin or estrual oviduct fluid. Incubation of sperm with 25% oviduct fluid collected at various days over the estrous cycle demonstrated that peak capacitating activity was found at estrus but was also present +/- 1 day from estrus. The active capacitating factor in oviduct fluid was found to be heat stable. In addition, when extraction procedures were applied in sequential order, oviduct fluid capacitating activity was resistant to protease digestion, precipitable by ethanol, size-excluded by Sephadex G-25, and destroyed by nitrous acid. These results suggest that a heparin-like glycosaminoglycan from the oviduct is a potential in vivo capacitating agent in the bovine.
对从长期插管的小母牛输卵管中收集的输卵管液进行了体外对牛精子获能作用的评估。通过精子在体外使牛卵母细胞受精的能力以及在暴露于溶血磷脂酰胆碱（LC）时发生顶体反应（AR）的能力来确定获能情况。将精子与0 - 25%（v/v）发情期输卵管液（发情期前后1天收集）孵育4小时后，再添加LC（100微克/毫升）额外孵育0.25小时，结果显示随着输卵管液浓度的增加，顶体反应精子的百分比也增加。与仅在培养基中孵育的精子相比，在25%发情期输卵管液中孵育4小时的精子使更多的卵母细胞受精（p < 0.05），但与在10微克/毫升肝素中孵育的精子无差异（p > 0.05）。葡萄糖抑制了LC诱导在肝素或发情期输卵管液中孵育4小时的精子发生AR的能力。在发情周期的不同天数收集25%输卵管液孵育精子表明，获能活性在发情期达到峰值，但在发情期前后1天也存在。发现输卵管液中的活性获能因子对热稳定。此外，当按顺序应用提取程序时，输卵管液的获能活性对蛋白酶消化有抗性，可被乙醇沉淀，被Sephadex G - 25排阻大小，且被亚硝酸破坏。这些结果表明，来自输卵管的一种肝素样糖胺聚糖是牛体内潜在的获能剂。
