RECEPTOR DYNAMICS DURING FERTILIZATION

受精过程中的受体动态

• 批准号: 2200311
• 负责人: RICHARD A CARDULLO
• 金额: $ 13.12万
• 依托单位: UNIVERSITY OF CALIFORNIA RIVERSIDE
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-08-01 至 1996-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2200311
• 关键词: digital imaging; egg /ovum; fertilization; fluorimetry; glycopeptides; glycoproteins; laboratory mouse; membrane activity; monoclonal antibody; receptor; receptor binding; receptor coupling; sperm

The overall goal of this proposal is to investigate the dynamics of sperm surface zona pellucida receptors (ZPrs) during fertilization. Fluorescently and isotopically labeled zona pellucida glycoproteins and glycopeptides will be used to determine: (1) the distribution of different ZPrs during sperm development, (2) the binding properties of ZPrs during fertilization, and (3) the sequence of cell surface events requisite for fertilization. Rather than using standard biochemical techniques requiring large populations of sperm, state-of-the-art digital imaging techniques will be used to determine the distributions and dynamics of zona pellucida receptors on the sperm surface. Using these methods, the investigator will observe the dynamics of sperm surface ZPrs during epididymal maturation, capacitation, and the acrosome reaction. A number of fluorescent substrates will be used for probing active sites of putative ZPrs on the surface of single sperm in well defined physiological states. Finally, a number of monoclonal and polyclonal antibodies will be used to follow the sequence of various cell surface molecules that are involved in the fertilization pathway. This research will yield important information about the distribution and dynamics ZPrs necessary for sperm-zona adhesion and induction of the acrosome reaction and may have implications both for the development of contraceptives and for understanding infertility problems involving sperm adhesion to the egg. In addition, elucidating the molecular dynamics of sperm adhesion to the zona pellucida will lead to a model of ligand-induced, receptor-mediated events important in many areas of cell biology.

该提议的总体目标是研究精子的动力 受精过程中表面Zona pellucida受体（ZPRS）。 荧光和同位素标记为Zona pellucida糖蛋白和 糖肽将用于确定：（1）不同的分布 精子发育过程中的ZPR，（2）ZPRS的结合特性 受精，（3）细胞表面事件的序列 受精。 而不是使用需要的标准生化技术 大量精子，最先进的数字成像技术 将用于确定Zona Pellucida的分布和动力学 精子表面的受体。 使用这些方法，研究者将 观察附睾成熟过程中精子表面ZPR的动力学， 电容和Adrosom反应。 许多荧光 基板将用于探测假定ZPRS的活动位点 明确定义的生理状态中的单个精子表面。 最后，一个 单克隆和多克隆抗体的数量将用于遵循 涉及各种细胞表面分子的序列 受精途径。 这项研究将产生重要信息 关于精子粘附所需的分布和动态ZPRS 和诱导ACROSOM反应，并且可能对 避孕药具的发展和理解不育 涉及精子粘附的问题。 另外，阐明 精子粘附的分子动力学对Zona pellucida将导致 配体诱导的，受体介导的事件在许多领域很重要 细胞生物学。