CONTROL OF CELLS OF THE REPRODUCTIVE AXIS

生殖轴细胞的控制

• 批准号: 6588487
• 负责人: BERTIL HILLE
• 金额: $ 15.33万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-04-01 至 2003-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6588487
• 关键词: adenylate cyclase; bicarbonates; biological signal transduction; calcium flux; cyclic AMP; fluorescence resonance energy transfer; germ cells; gonadotropin releasing factor; immunocytochemistry; laboratory mouse; laboratory rat; membrane potentials; phosphorylation; pituitary gland; protein kinase; single cell analysis; spectrometry; sperm; sperm motility; spermatogenesis; testis; tissue /cell culture; voltage /patch clamp; western blottings

This project concerns signaling mechanisms in cells of the reproductive axis. Electrical excitability and rat intracellular signaling will be studied in male mouse germ-line cells and epididymal spermatozoa and in male rat pituitary gonadotropes. The physiology of single living cells will be studied by patch clamp and optical indicators. The guiding hypotheses are that membrane potential, intracellular Ca2+, and cyclic AMP are key cellular signals that control steps in reproduction. The ion channels of spermatids and sperm will be identified by patch clamp and immunocytochemistry, with special attention to voltage-gated Ca2+ channels. In sperm and spermatids the actions of bicarbonate on cyclic AMP, protein phosphorylation, motility and ion channel function will be compared. Mechanisms regulating Ca2+ clearance from sperm and spermatid cytoplasm will be identified. In gonadotropes, immunocytochemical experiments will identify the IP3 uptake and buffering of Ca2+ to permit formulation and test of kinetic models for cellular Ca2+ dynamics during the Ca2+ oscillations induced by gonadotropin releasing hormone. These studies will identify potential targets for future male contraceptives.

该项目涉及生殖轴细胞中的信号传导机制。将在雄性小鼠生殖细胞和附睾精子以及雄性大鼠垂体促性腺激素中研究电兴奋性和大鼠细胞内信号传导。将通过膜片钳和光学指示器来研究单个活细胞的生理学。指导性假设是膜电位、细胞内 Ca2+ 和环 AMP 是控制繁殖步骤的关键细胞信号。将通过膜片钳和免疫细胞化学来识别精细胞和精子的离子通道，特别关注电压门控 Ca2+ 通道。在精子和精细胞中，碳酸氢盐对环腺苷酸、蛋白质磷酸化、运动性和离子通道功能的作用将进行比较。将确定调节精子和精细胞质中 Ca2+ 清除的机制。在促性腺激素中，免疫细胞化学实验将鉴定 IP3 摄取和 Ca2+ 缓冲，以允许在促性腺激素释放激素诱导的 Ca2+ 振荡期间制定和测试细胞 Ca2+ 动力学的动力学模型。这些研究将确定未来男性避孕药的潜在目标。