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.

男性和女性因素导致的不孕症发病率持续上升 尽管生殖技术取得了许多进步，但仍会上升。一些 人类配子相互作用的异常已被证明是由于 精子缺陷，而其他缺陷已被证明是 ZP 缺陷。我们的 缺乏对相互作用所涉及的分子机制的了解 与不育女性相比，具有可育 ZP 的人类精子的数量 由于人类卵母细胞的缺乏，男性的数量受到限制 实验研究的伦理限制。最近的进展使用 体外受精技术，包括细胞内精子注射 (ICSI) 直接进入卵细胞溶胶已被证明是有效的 许多案例，尽管伦理和临床方面的担忧日益增加 到这项技术的普遍使用。它正变得越来越 显然，改进的临床检测对于评估精子是必要的。 ZP 互动，以评估成功的最佳程序 受精和怀孕。鉴于分子生物学的最新进展 生物学上，编码三种主要人类 ZP 蛋白的基因已被 鉴定和 cDNA 可用于开始更好地评估 精子-ZP 相互作用的分子基础。因为 ZP 蛋白 不同的哺乳动物物种已被证明在 在受精过程中，显然人类 ZP 蛋白必须 用于评估人类精子-ZP 相互作用。具体来说，就是 建议 (1) 使用以下方法表达和表征三种人类 ZP 蛋白 原核和真核表达系统的差异 糖基化糖蛋白； (2) 确定这些表达的功能 ZP 蛋白在精子结合和顶体反应启动中的作用 剖析所涉及的 ZP 蛋白的功能域：以及 (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 参与精子/ZP 相互作用和发育的分子异常 评估不育男性的临床测定。 (4) 判断是否有 不孕女性的 ZP 蛋白存在缺陷，可以使用以下方法进行识别： 单链构象多态性。这些研究还应该允许 我们定义参与这些相互作用的分子，这将有助于 治疗不孕症并提供重要信息 开发改进的避孕方法。