MINORITY PREDOCTORAL FELLOWSHIP PROGRAM

描述（由申请人提供）：我们的目标是阐明精子超活化中涉及的信号通路。超活化，一种活跃的精子活动类型，在受精时在输卵管中发现。由于精子的超活化能力通常是精子生育力的指标，因此确定超活化运动力调节的机制可以导致开发促进精子生育力的治疗方法，或者导致开发针对精子运动力特异性调节剂的避孕药。我们的具体目标是使用普鲁卡因诱导的过度激活的公牛精子，以确定的作用：1）蛋白质磷酸化在过度激活的精子使用蛋白质印迹分析和间接免疫荧光。2)细胞内Ca 2+在超激活运动中的荧光Ca 2+指示剂和单细胞成像。3)通过使用pH敏感荧光探针和单细胞图像分析测量细胞内pH来调节超活化运动的细胞质碱化。我们建议，超激活是由一个信号通路介导的鞭毛蛋白磷酸化，这是依赖于Ca2+内流和碱化的胞质溶胶。

DESCRIPTION (provided by applicant): Our goal is to elucidate the signaling pathway involved in sperm hyperactivation. Hyperactivation, a vigorous type of sperm motility, is found in the oviduct at time of fertilization. Because the ability of sperm to hyperactivate is often an indictor of sperm fertility, determining the mechanism by which hyperactivated motility is regulated can lead to development of treatments to promote sperm fertility or alternatively, result in development of contraceptives targeting specific modulators of sperm motility. Our specific aims are to use procaine-induced hyperactivated bull sperm to determine the role of: 1) Protein phosphorylation in hyperactivated sperm using western blot analysis and indirect immunofluorescence. 2) Intracellular Ca 2+ in hyperactivated motility using fluorescent Ca 2+ indictors and single cell imaging. 3) Cytoplasmic alkalization in modulating hyperactivated motility by measuring intracellular pH using pH sensitive fluorescent probes and single cell image analysis. We propose that hyperactivation is mediated by a signaling pathway involving phosphorylation of flagellar proteins, which is dependent on Ca 2+ influx and alkalization of the cytosol.