RECEPTOR DYNAMICS DURING FERTILIZATION

这项提议的总体目标是研究精子的动力学。 受精过程中的表面透明带受体(ZPR)。 荧光和同位素标记的透明带糖蛋白和 糖肽将被用来确定：(1)不同的分布 精子发育过程中的ZPrs；(2)精子发育过程中ZPrs的结合特性 受精，以及(3)细胞表面事件的顺序 受精。而不是使用标准的生化技术，需要 大量的精子，最先进的数字成像技术 将用于确定透明带的分布和动态 精子表面的受体。使用这些方法，调查员将 观察精子表面ZPR在附睾成熟过程中的动态变化， 获能和顶体反应。一批荧光粉 底物将被用来探测假定的ZPR在 单个精子在明确的生理状态下的表面。最后，一个 将使用大量的单抗和多克隆抗体来跟踪 各种细胞表面分子的序列，这些分子参与了 受精途径。这项研究将提供重要的信息 精子-小带黏附所需ZPR的分布和动态 和顶体反应的诱导，并可能对 避孕药具的发展和对不孕症的认识 涉及精子与卵子粘连的问题。此外，澄清 精子与透明带黏附的分子动力学将导致 配体诱导、受体介导的事件模型在许多领域都很重要 细胞生物学的研究。

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.