MINORITY PREDOCTORAL FELLOWSHIP PROGRAM

描述（由申请人提供）：我们的目标是阐明与精子过度激活有关的信号通路。过度激活，一种活跃的精子运动类型，在受精时发现于输卵管。由于精子过度激活的能力通常是精子生育能力的一个指标，因此确定过度激活的运动被调节的机制可以导致促进精子生育的治疗的发展，或者导致针对精子运动的特定调节剂的避孕药的发展。我们的具体目的是使用普鲁卡因诱导的高激活公牛精子来确定：1)蛋白质磷酸化在高激活精子中的作用，使用western blot分析和间接免疫荧光。2)利用荧光ca2 +指标和单细胞成像技术研究细胞内ca2 +在超激活运动中的作用。3)利用pH敏感荧光探针和单细胞图像分析测量细胞内pH值，细胞质碱化在调节过度激活运动中的作用。我们提出，过度激活是由鞭毛蛋白磷酸化的信号通路介导的，该信号通路依赖于ca2 +内流和细胞质溶胶的碱化。

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.