Oviduct-specific glycoprotein and heparin modulate sperm-zona pellucida interaction during fertilization and contribute to the control of polyspermy
Oviduct-specific glycoprotein and heparin modulate sperm-zona pellucida interaction during fertilization and contribute to the control of polyspermy
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DOI：
10.1073/pnas.0804422105
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发表时间：
2008-10-14
影响因子：
11.1
通讯作者：
Aviles, Manuel
中科院分区：
文献类型：
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作者：
Coy, Pilar;Canovas, Sebastian;Aviles, Manuel
多精受精是胎盘哺乳动物的一种重要的受精异常，会导致胚胎过早死亡。它在体外条件下尤其常见，使成功产生可存活胚胎变得复杂化。由于精子进入后的变化（即皮质颗粒胞吐作用），多精受阻发生。然而，其他因素可能在精子与卵母细胞相互作用之前作用于配子，从而在调节多精受精方面发挥重要作用。大多数研究都使用啮齿类动物作为模型，但有蹄类动物在防止多精受精的机制上可能有所不同。我们假设卵母细胞沿输卵管壶腹部运输过程中透明带（ZIP）发生变化，调节与精子的相互作用，从而有助于多精受精的调节。我们在此报告，母猪和小母牛的排卵期输卵管液 (OF) 增加了（反相和异源性）ZIP 对链霉蛋白酶（通常用于测量多精受阻的参数）消化的抗性，消化时间从大约 1 分钟（猪）或 2 分钟（牛）变为 45 分钟（猪）或几个小时（牛）。在这两个物种中，卵母细胞暴露于 OF 会增加体外受精后的单精子性，而在猪中，精子与 ZP 的结合会减少。 OF暴露的卵母细胞对链霉蛋白酶的抵抗力通过暴露于肝素耗尽的培养基而被消除；在含有肝素的培养基中，它没有改变。去除 OF 暴露的卵母细胞后，对肝素耗尽的培养基中释放的内容物进行蛋白质组学分析，从而可以分离和鉴定输卵管特异性糖蛋白。因此，输卵管特异性糖蛋白-肝素蛋白复合物似乎与受精前输卵管 ZIP 变化有关，影响精子结合并有助于多精受精的调节。
Polyspermy is an important anomaly of fertilization in placental mammals, causing premature death of the embryo. It is especially frequent under in vitro conditions, complicating the successful generation of viable embryos. A block to polyspermy develops as a result of changes after sperm entry (i.e., cortical granule exocytosis). However, additional factors may play an important role in regulating polyspermy by acting on gametes before sperm-oocyte interaction. Most studies have used rodents as models, but ungulates may differ in mechanisms preventing polyspermy. We hypothesize that zona pellucida (ZIP) changes during transit of the oocyte along the oviductal ampulla modulate the interaction with spermatozoa, contributing to the regulation of polyspermy. We report here that periovulatory oviductal fluid (OF) from sows and heifers increases (both, con- and heterospecifically) ZIP resistance to digestion with pronase (a parameter commonly used to measure the block to polyspermy), changing from digestion times of approximate to 1 min (pig) or 2 min (cattle) to 45 min (pig) or several hours (cattle). Exposure of oocytes to OF increases monospermy after in vitro fertilization in both species, and in pigs, sperm-ZP binding decreases. The resistance of OF-exposed oocytes to pronase was abolished by exposure to heparin-depleted medium; in a medium with heparin it was not altered. Proteomic analysis of the content released in the heparin-depleted medium after removal of OF-exposed oocytes allowed the isolation and identification of oviduct-specific glycoprotein. Thus, an oviduct-specific glycoprotein-heparin protein complex seems to be responsible for ZIP changes in the oviduct before fertilization, affecting sperm binding and contributing to the regulation of polyspermy.