THE CELL BIOLOGY OF ASCIDIAN SPERM ACTIVATION

这个项目的目标是增加参与的少数民族的数量。 在研究生育力和生物医学问题时 受精、跨膜信号传递和细胞内运动 细胞器通过研究激活的海囊精子在细胞和 生化水平。在我的学校工作过的学生中有30% 实验室在过去的十年里一直是少数族裔。这些学生中有许多 已经进入了医疗行业，但很少有人继续从事生物医学 研究。我实验室的学生参与MBR计划 会增加他们对生物医学研究的接触，作为一种可行的职业 选择。这些学生将研究受精的早期事件 包括精子与卵子结合、精子激活以及随后的精子 鸡蛋衣物的渗透，最终导致融合的过程 精子和卵子质膜。精子的细胞机制 精子-卵子结合所触发的激活和穿透是 这个项目的重点。据推测，信号事件 跨膜必须将结合和激活联系起来， 产生作用力的细胞骨架元素的重排必须连接 激活到线粒体迁移。研究跨膜 信号连接需要用激活剂或阻滞剂加载细胞 要间接测试是否存在 聚磷脂酰肌醇连接机理。将收集直接证据 通过对该途径的一些蛋白质成员进行生化分析，通过 测量他们行为的产品的外观，并通过研究 它们在抗体存在时的行为，这些抗体识别或 干扰他们。这种聚磷酰肌醇依赖的调节 之所以选择途径，是因为在一些细胞中，它被认为可以提高 细胞内pH和钙，两者都是海鞘的一部分 精子激活。研究线粒体运动需要探索 参与细胞骨架的生化性质和细胞定位 蛋白质。特定细胞骨架蛋白的存在与运动性 电机将通过聚丙烯酰胺凝胶电泳法和 免疫印迹。这些蛋白质的细胞定位将被研究。 通过间接免疫荧光和免疫电子显微镜观察。

The goal of this project is to increase the number of minorities involved in research addressing the biomedical problems of fertility and fertilization, signaling across membrane, and intracellular movement of organelles via studies of activated ascidian sperm at the cellular and biochemical levels. Thirty percent of the students who have worked in my' lab over the past ten years have been minorities. Many of these students have gone into health professions, but few have continued in biomedical research. The participation of students in my lab in the MBRS program would increase their exposure to biomedical research as a viable career option. These students would be studying the early events of fertilization include sperm-egg binding, sperm activation and, subsequently, sperm penetration of egg vestments, processes which ultimately lead to fusion of sperm and egg plasma membranes. The cellular mechanisms of sperm activation and penetration, as triggered by sperm-egg binding, are the foci of this project. It is hypothesized that signaling events which cross the membrane must connect binding to activation and that rearrangement of force-producing cytoskeletal elements must connect activation to mitochondrial migration. Investigating the transmembrane signaling connection requires loading of cells with activators or blockers to indirectly test for the presence of the components of a polyphosphainositide-linked mechanism. Direct evidence will be gathered by biochemical analysis of some of the protein members of the pathway, by measuring the appearance of the products of their actions, and by studying their behavior in the presence of antibodies which either identify or interfere with them. This polyphosphainositide-dependent regulatory pathway was chosen because, in some cells, it is known to elevate intracellular pH and calcium, both of which occur as part of ascidian sperm activation. Studying mitochondrial movement requires probing the biochemical nature and cellular location of participating cytoskeletal proteins. The presence of specific cytoskeletal proteins and motility motors will be established by polyacrylamide gel electrophoresis and immunoblotting. The cellular location of these proteins will be studied by indirect immunofluorescence and immunoelectron microscopy.