MINORITY PREDOCTORAL FELLOWSHIP PROGRAM

少数族裔博士前奖学金计划

基本信息

批准号：
6591093
负责人：
BECKY MARQUEZ
金额：
$ 3.37万
依托单位：
CORNELL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-05-16 至 2005-05-15
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6591093
关键词：
SDS polyacrylamide gel electrophoresis acidity /alkalinity bioimaging /biomedical imaging biological signal transduction calcium flux cyclic AMP cytoplasm fertility phosphorylation predoctoral investigator procaine protein kinase A single cell analysis sperm sperm motility video microscopy western blottings

项目摘要

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.

描述（由申请人提供）：我们的目标是阐明精子过度激活所涉及的信号通路。受精时在输卵管中发现了过度活跃，这是一种精子活力旺盛的现象。由于精子过度活跃的能力通常是精子生育力的指标，因此确定过度活跃运动的调节机制可以导致开发出促进精子生育力的治疗方法，或者导致开发针对精子运动特定调节剂的避孕药。我们的具体目标是使用普鲁卡因诱导的过度活跃的公牛精子来确定以下作用：1) 使用蛋白质印迹分析和间接免疫荧光法确定过度活跃的精子中的蛋白质磷酸化。 2) 使用荧光 Ca 2+ 指示器和单细胞成像观察细胞内 Ca 2+ 运动过度。 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来调节过度活跃的运动的细胞质碱化。我们认为过度激活是由涉及鞭毛蛋白磷酸化的信号通路介导的，该信号通路依赖于 Ca 2+ 流入和胞质溶胶的碱化。