MOLECULAR APPROACHES FOR EVAULATION OF HUMAN FERTILITY

评估人类生育能力的分子方法

• 批准号: 6440496
• 负责人: BONNIE S DUNBAR
• 金额: $ 17.42万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-04-01 至 2002-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6440496
• 关键词: cell cell interaction; egg /ovum; fertility; gene expression; laboratory rabbit; molecular biology; reproductive system disorder; sperm analysis; zona pellucida glycoproteins

The incidence of infertility due to both male and female factors continues to rise despite many advances in reproductive technologies. Some abnormalities in human gamete interaction have been shown to be due to defects in the sperm while others have been shown to be ZP defects. Our lack of understanding of molecular mechanisms involved in the interaction of human sperm with the ZP in fertile as compared to infertile females and males has been limited due to the unavailability of human oocytes and ethical restraints for experimental studies. Recent advances using in vitro fertilization technology which include intracellular sperm injection (ICSI) directly into the egg cytosol have been shown to be effective in many cases, although there are increasing ethical and clinical concerns as to the universal use of this technology. It is becoming increasingly apparent that improved clinical assays are necessary for evaluating sperm- ZP interaction in order to assess the optimal procedures for successful fertilization and pregnancy. In view of recent advances in molecular biology, the genes encoding the three major human ZP proteins have been identified and cDNAs are available to begin to better evaluate the molecular basis of sperm-ZP interaction. Because the ZP proteins of different mammalian species have been shown to have distinct roles in the fertilization process, it is apparent that the human ZP proteins must be utilized to evaluate human sperm- ZP interaction. Specifically, it is proposed to (1) express and characterize the three human ZP proteins using prokaryotic and eukaryotic expression systems that differentially glycosylate glycoproteins; (2) identify the functions of these expressed ZP proteins in sperm binding and initiation of the acrosome reaction and dissect the functional domains of the ZP proteins involved: and (3) use sperm from infertile males to determine if sperm dysfunction is due to molecular abnormalities involved in sperm/ZP interaction and develop clinical assays to evaluate infertile males. (4) determine if some infertile women have defects in ZP proteins that can be identified using single strand conformational polymorphism. These studies should also allow us to define molecules involved in these interactions which will aid in the treatment of infertility as well as provide information critical for the development of improved contraceptive methods.

由于男性和女性因素引起的不育症的发生率仍在继续 尽管生殖技术取得了许多进步，但仍在上升。一些 人配子互动中的异常已被证明是由于 精子中的缺陷，而其他人则被证明是ZP缺陷。我们的 对相互作用涉及的分子机制缺乏了解 与不育女性相比，人类精子具有肥沃的ZP 由于人类卵母不可用，男性受到限制 实验研究的道德约束。最近使用的进步 体外受精技术，包括细胞内精子注射 （ICSI）直接显示出卵子醇在 许多情况，尽管越来越多的道德和临床问题是 为了普遍使用这项技术。它变得越来越 显然，改善临床测定对于评估精子是必要的 ZP相互作用以评估成功的最佳程序 受精和怀孕。鉴于分子的最新进展 生物学，编码三种主要人ZP蛋白的基因已经 确定的和CDNA可以开始更好地评估 精子相互作用的分子基础。因为 已经显示出不同的哺乳动物物种在 受精过程，显然人类ZP蛋白必须是 用于评估人类精子的相互作用。具体来说，是 提出（1）使用三种人ZP蛋白来表征（1） 原核生物和真核表达系统差异 糖基化糖蛋白； （2）确定这些表达的功能 ZP蛋白质中的精子结合和启动Adrosom反应和开始 剖析所涉及的ZP蛋白的功能域：（3）使用 来自不育男性的精子以确定精子功能障碍是否是由于 精子/ZP相互作用涉及的分子异常并发展 评估不育男性的临床测定法。 （4）确定是否有些 不育妇女在ZP蛋白中具有缺陷，可以使用 单链构象多态性。这些研究也应允许 我们定义参与这些相互作用的分子，这将有助于 不育的治疗以及提供至关重要的信息 改进的避孕方法的发展。