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。将在男性小鼠系细胞和附睾精子和雄性大鼠垂体性促性腺激素中研究电兴奋性和大鼠细胞内信号传导。单个活细胞的生理学将通过斑块夹和光学指标研究。引导假设是膜电位，细胞内CA2+和环状AMP是控制繁殖步骤的钥匙细胞信号。精子和精子的离子通道将通过斑块夹和免疫细胞化学识别，并特别注意电压门控Ca2+通道。在精子和精子中，将比较碳酸氢盐对环状AMP的作用，将比较蛋白质磷酸化，运动能力和离子通道功能。将确定调节精子和精子细胞质的Ca2+清除率的机制。在促性腺激素中，免疫细胞化学实验将识别Ca2+的IP3摄取和缓冲，以允许在促性腺激素释放激素引起的Ca2+振荡期间对细胞Ca2+动力学的动力学模型进行制定和测试。这些研究将确定未来男性避孕药的潜在靶标。