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Studies on the mechanism of sperm-egg interaction

精卵相互作用机制研究

基本信息

批准号：
59540475
负责人：
SHIMIZU Satoru
金额：
$ 1.47万
依托单位：
Aichi Cancer Center Research Institute
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1984
资助国家：
日本
起止时间：
1984 至 1986
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-59540475/
关键词：
zona pellucida glycoprotein 精子リセプター

项目摘要

In the present study, we analyzed the sperm receptor activity of the zona glycoproteins from mouse ovarian oocytes, and oligosaccharides in individual glycoproteins. For the determination of the glycoprotein that has the sperm binding activity, we used a in vitro binding assay. Capacitated sperm were tested for the binding to the glycoprotein immobilized on latex beads. The results indicated that ZP-3 glycoprotein had the highest capacity to bind sperm. However, no positive results were obtained in an attempt to determine the sperm components that had the binding activity to ZP-3.Assuming that the ZP-3 glycoprotein might have the unique carbohydrates reflecting its functional difference from the rest of zona glycoproteins, gel chromatographic comparison of the oligosaccharides was made using isotopically labeled glycoproteins. All of the zona glycoproteins contained both O- and N-linked oligosaccharides, although carbohydrate content and ratio of the amount of O- and N-linked oligosaccharides siffered among the glycoproteins. N-linked carbohydrates had negative charge not released by sialidase, and contained N-acetylgalactosamine usally not found in N-linked oligosaccharides. All three glycoproteins showed the similar patterns on gel chromatography of both O- and N-linked oligosaccharides, and no characteristic difference of oligosaccharides has been detected among the glycoproteins. However, ZP-3 oligosaccharides from two-cell embryos gave altered patterns from unfertilized eggs, probably reflecting the functional modification of the zona pellucida in two-cell embryos. Future studies on these oligosaccharides that undergo the structural changes in fertilization could result in the elucidation of the mechanism of sperm-egg interaction.

在本研究中，我们分析了小鼠卵巢卵母细胞的精子受体活性的糖蛋白，和寡糖的个别糖蛋白。为了确定具有精子结合活性的糖蛋白，我们使用体外结合试验。测试获能精子与固定在乳胶珠上的糖蛋白的结合。结果表明，ZP-3糖蛋白结合精子的能力最强。然而，在试图确定与ZP-3具有结合活性的精子组分时，没有得到阳性结果。假设ZP-3糖蛋白可能具有反映其与其他精子糖蛋白功能差异的独特碳水化合物，用同位素标记的糖蛋白进行寡糖的凝胶色谱比较。所有的糖蛋白都含有O-和N-连接的寡糖，尽管糖蛋白中的碳水化合物含量和O-和N-连接的寡糖的量的比率是不同的。N-连接碳水化合物具有不被唾液酸酶释放的负电荷，并且含有通常在N-连接寡糖中不存在的N-乙酰半乳糖胺。所有三种糖蛋白的O-和N-连接的寡糖的凝胶色谱显示出相似的模式，并没有检测到糖蛋白之间的寡糖的特征性差异。然而，ZP-3寡糖从两细胞胚胎未受精卵的模式发生了变化，可能反映了在两细胞胚胎的透明质酸的功能修改。未来对这些在受精过程中发生结构变化的寡糖的研究可能会导致精卵相互作用机制的阐明。