HUMAN SPERM-ZONA PELLUCIDA INTERACTIONS

人类精子与透明带的相互作用

• 批准号: 3470514
• 负责人: ROBERT B SHABANOWITZ
• 金额: $ 9.74万
• 依托单位: GEISINGER CLINIC
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-05-01 至 1996-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3470514
• 关键词: active sites; antibody formation; binding proteins; cell cell interaction; cross immunity; egg /ovum; electrophoresis; extracellular matrix; fertilization; fluorescence microscopy; human subject; laboratory mouse; laboratory rabbit; monoclonal antibody; sperm

In mammalian fertilization, one important site of sperm-egg interaction occurs on the surface of the zona pellucida (ZP). The ZP is an extracellular matrix specific to mature mammalian oocytes. Sperm must first attach to and penetrate the ZP prior to actual fertilization of the egg. Specific receptors for sperm-ZP recognition are believed to reside on each gamete. Human sperm-ZP interactions will be studied by an examination of the specific sperm and ZP molecules that are directly involved in sperm-zona recognition and binding. Rabbit polyclonal antibodies will be raised against human sperm antigens which have previously been shown to have affinity for the ZP. These antibodies will be used to localize the sperm antigens on the sperm membrane by epifluorescent microscopy. Localization of the antigens on the sperm membrane will provide information on the regionalization of these antigens. The antibodies will also be used to examine whether or not they can inhibit sperm-zona interaction in vitro. Parallel studies on the human ZP will involve electrophoretic and lectin binding characteristics of the ZP molecules. Monoclonal antibodies (MAB) will be produced against solubilized human ZP and screened against the individual molecules that comprise the ZP to obtain MAB's specific for each ZP component. The MAB's will be used to examine their ability to interfere with sperm-ZP binding, and to localize regions on the human sperm which bind specific ZP components. The cross-reactivity of antiZP and anti-sperm antibodies with sperm and ZP of other mammalian species will be examined to determine if the specific molecules involved in sperm-ZP recognition are common among species. Naturally occurring anti-sperm and anti-ZP antibodies have been shown to interfere with fertilization. The identification and characterization of the specific molecules involved, therefore, may not only aid in the diagnosis of also have contraceptive potential as well.

在哺乳动物受精过程中，精子与卵子相互作用的一个重要部位 发生在透明带(ZP)表面。ZP是一个 成熟的哺乳动物卵母细胞特有的细胞外基质。精子必须 在实际受精之前，首先附着并穿透ZP 蛋。精子-ZP识别的特定受体被认为驻留在 每一个配子。将通过一项检查来研究人类精子与ZP的相互作用 直接参与的特定精子和ZP分子的 精子带识别和结合。兔多克隆抗体将成为 针对人类精子抗原而产生的抗原，此前已表明 对ZP有亲和力。这些抗体将被用来定位 荧光显微镜观察精子膜上的精子抗原。 精子膜上抗原的定位将提供信息 关于这些抗原的区域化。抗体也将被用于 以检测它们是否能抑制精子与卵泡的相互作用 体外培养。对人类ZP的平行研究将涉及电泳法和 凝集素与ZP分子的结合特性。单克隆抗体 (MAb)将针对溶解的人ZP生产并筛选 组成ZP的单个分子，以获得MAB的特异性 每个ZP组件。人与生物圈计划将被用来检验他们的能力 干扰精子与ZP的结合，并定位人类精子上的区域 它绑定了特定的ZP组件。抗ZP与ZP的交叉反应性 与其他哺乳动物的精子和ZP一起产生的抗精子抗体 检查以确定精子ZP中是否涉及特定分子 在物种中识别是很常见的。自然产生的抗精子和 抗ZP抗体已被证明会干扰受精。这个 对涉及的特定分子进行鉴定和表征， 因此，可能不仅有助于诊断的也有避孕药 也很有潜力。