RECEPTOR DYNAMICS DURING FERTILIZATION

这项提案的总体目标是调查精子的动力学 受精过程中的表面透明质酸受体（ZPrs）。 同位素标记的透明质酸糖蛋白， 糖肽将用于确定：（1）不同糖肽的分布 ZPrs在精子发育过程中的结合特性，（2）ZPrs在精子发育过程中的结合特性。 受精，和（3）所需的细胞表面事件的顺序， 受精 而不是使用标准的生化技术， 大量的精子，最先进的数字成像技术 将用于确定透明珊瑚的分布和动态 精子表面的受体。 使用这些方法，研究人员将 观察附睾成熟过程中精子表面ZPrs的动态变化， 获能和顶体反应。 的多个荧光 底物将用于探测假定的ZPr在 单个精子在明确定义的生理状态下的表面。 最后 将使用许多单克隆和多克隆抗体来跟踪 这些细胞表面分子的序列参与了 受精途径 这项研究将产生重要的信息 关于精子粘附所必需的ZPrs的分布和动态 和诱导顶体反应，并可能对 避孕药的发展和了解不孕症 精子与卵子的粘附问题。 此外，阐明 精子粘附于透明膜的分子动力学将导致 配体诱导的受体介导的事件模型，在许多领域都很重要 细胞生物学。

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.