CARBOHYDRATE DETERMINANTS IN MAMMALIAN FERTILIZATION

哺乳动物受精中的碳水化合物决定因素

• 批准号: 2561902
• 负责人: RICHARD A CARDULLO
• 金额: $ 7.26万
• 依托单位: UNIVERSITY OF CALIFORNIA RIVERSIDE
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-03-01 至 2000-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2561902
• 关键词: acrosome; affinity chromatography; carbohydrate receptor; carbohydrates; cell cell interaction; egg /ovum; fertilization; glycolipids; laboratory mouse; mass spectrometry; monosaccharides; oligosaccharides; receptor binding; tissue /cell culture; zona pellucida glycoproteins

DESCRIPTION: (Adapted from applicant's abstract) The overall goal of this proposal is to determine the specific carbohydrate determinants within the zona pellucida that are recognized by mammalian sperm during fertilization. A number of biochemical and cell biological approaches will be used to precisely determine the exact nature of the oligosaccharides on the glycoprotein ZP3, which are necessary for sperm-zone adhesion and the induction of acrosomal exocytosis. Quantitative assays using solubilized and 125I-labeled zone pellucida glycoproteins to characterize the interaction between ZP3 and its complementary receptor(s) on the mouse sperm surface have been developed. These assays will be used to probe the specific molecular interactions involved in the recognition between sperm and egg in a manner which is not possible using previous cell binding assays. Using this new assay, defined monosaccharides, simple oligosaccharides, and complex oligosaccharides will be compared to identify potential carbohydrate moieties which are involved in sperm adhesion to the zona pellucida. Specifically, oligo-saccharides which contain terminal sugars that have been implicated in sperm-zone adhesion will be tested. By varying both the composition and the complexity of these oligosaccharides, the carbohydrate recognition domains on the sperm's ZP3 receptor will be examined. In addition, the ZP3 glycoprotein will be chemically dissected to quantify the contributions of both O-linked and N-linked oligosaccharides, as well as specific terminal monosaccharides on ZP3, in sperm-zone binding. These oligosaccharides will be initially analyzed using lectin affinity chromatography and mass spectrometry to determine the specific sugars and oligosaccharide mass in the ZP3 gly-coprotein. This research will yield important information about the composition of macromolecules involved in mammalian sperm-egg recognition and adhesion and may have implications both for the development of contraceptives and for understanding specific molecular lesions which may lead to infertility.

描述：（改编自申请人的摘要） 建议是确定特定的碳水化合物决定因素 在受精过程中由哺乳动物精子识别的Zona pellucida。 许多生化和细胞生物学方法将用于 精确确定寡糖在 糖蛋白ZP3，对于精子粘附和 诱导诊断性胞吐作用。 使用溶解的定量测定 和125i标记的区域pellucida糖蛋白，以表征 ZP3与其互补受体之间的相互作用在小鼠精子上 表面已经开发。 这些测定将用于探测 精子之间识别的特定分子相互作用 和鸡蛋以先前的细胞结合不可能的方式 测定。 使用这种新测定，定义的单糖，简单 将比较寡糖和复杂的寡糖以鉴定 潜在的碳水化合物部分与精子粘附有关 Zona Pellucida。 具体而言，含有终端的寡糖 将测试与精子粘附有关的糖。 经过 改变这些寡糖的组成和复杂性， 精子ZP3受体上的碳水化合物识别域将是 检查。 此外，ZP3糖蛋白将化学剖析 量化O连锁和N连接的寡糖的贡献， 以及ZP3上的特定末端单糖，在精子区结合中。 这些寡糖最初将使用凝集素亲和力分析 色谱和质谱法，以确定特定的糖和 ZP3 Gly-Coprotein中的寡糖质量。 这项研究将产生 有关涉及大分子的组成的重要信息 哺乳动物的精子识别和粘附，可能都有含义 用于开发避孕药具和理解特定的 可能导致不育症的分子病变。