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Analysis of carbohydrate recognition mechanism in fertilization

受精过程中碳水化合物识别机制分析

基本信息

批准号：
10680578
负责人：
TAKASAKI Seiichi
金额：
$ 0.64万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10680578/
关键词：
gamete carbohydrate recognition system sperm egg fertilization シアロ糖鎖

项目摘要

It has been suggested that multiple carbohydrate recognition systems are working in the gamete interaction. In this project, carbohydrate structures of eggs recognized by boar sperm and carbohydrate recognition molecules on sperm were investigated on the basis of oligosaccharide structures of porcine zona pellucida glycoproteins, and the following results were obtained.1) Observation of sperm binding to fetuin-beads and their glycosidase digests showed that boar sperm recognize sialyl and non-sialyl N-acetyllactosamine structures.2) The sperm also adhered to LeィイD1xィエD1 oligosaccharide probe-coupled avidin-Sepharose beads, but not to beads carrying sialyl LeィイD1xィエD1 and type 2H oligosaccharides and Fuc-GlcNA.3) It was shown by the competition experiment that recognition of the LeィイD1xィエD1 structure by sperm is distinct from that of the sialyl and non-sialyl N-acetyllactosamine structures.4) Dextran-based multivalent oligosaccharide polymers carrying either of sialyl and non-sialyl N-acetyllactosamine and LeィイD1xィエD1 structures, which were prepared by our recently developped method, inhibited sperm binding to eggs.5) Cytochemical analysis using biotinylated multivalent oligosaccharide probes and fluorescent streptoavidin indicated that the heads of capacitated sperm are stained with either of the probes carrying sialyl and non-sialyl N-acetyllactosamine and LeィイD1xィエD1 structures, but those of acrosome-reacted sperm are not.These results suggest that multiple interactions between several carbohydrate recognition molecules on sperm and their oligosaccharide ligands on eggs are involved in fertilization.

研究表明，在配子相互作用中存在着多种糖类识别系统。本课题以猪卵透明质酸糖蛋白的寡糖结构为基础，研究了猪精子识别卵子的糖结构和精子上的糖识别分子，结果表明：1）猪精子与胎球蛋白结合的观察表明，猪精子能识别唾液酸化和非唾液酸化的N-乙酰乳糖胺结构;精子还粘附到Le BocyanD 1x BocyanD 1寡糖探针偶联的抗生物素蛋白-琼脂糖珠上，3）竞争实验表明，精子对Leu D1 x Leu D1结构的识别不同于对唾液酸化和非唾液酸化N-葡萄糖酸的识别。乙酰基乳糖胺结构。4）携带唾液酸和非唾液酸N-乙酰基乳糖胺和Le β D1 x β D1结构的葡聚糖基多价寡糖聚合物，其通过我们最近开发的方法制备，抑制精子与卵子结合。使用生物素标记的多价寡糖探针和荧光链霉亲和素进行的细胞化学分析表明，获能精子的头部是结果表明，精子上的糖识别分子与卵子上的寡糖配体之间存在着多种相互作用，精子上的糖识别分子与卵子上的寡糖配体之间存在着多种相互作用。