INTRACELLULAR CA2+ STORES AND THE IP3 RECEPTOR

细胞内 CA2 存储和 IP3 受体

• 批准号: 6106792
• 负责人: JAMES W PUTNEY
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ENVIRONMENTAL HEALTH SCIENCES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6106792
• 关键词: calcium; calcium binding protein; calcium channel; calcium flux; calcium indicator; carbohydrate receptor; confocal scanning microscopy; endoplasmic reticulum; inositol phosphates; intracellular; receptor expression; tissue /cell culture; video microscopy

Summary of Work: Calcium signalling in non-excitable cells generally involves mobilization of Ca2+ from intracellular stores by the second messenger, inositol 1,4,5-trisphosphate (IP3). However, although it is clear that the locus of this intracellular store of Ca2+ is either a part of, or is derived from endoplasmic reticulum, the precise locus of these calcium stores and their relationship to other cellular structures is not clear. In addition, the interaction of IP3 with its receptor on these Ca2+ stores often results in complex kinetics of Ca2+ release and reaccumulation, resulting in the appearance of regenerative cytoplasmic intracellular Ca2+ ([Ca2+]i) oscillations. These stores of Ca2+ are important because they are the initial source for the [Ca2+]i signals generated in many different cell types by the actions of hormones, neurotransmitters and growth factors. Changes in the concentration in these stores also initiates a novel signalling pathway that controls such processes as apoptosis, gene expression, and capacitative calcium entry. The focus of research in this project is to identify and localize the intracellular Ca2+ pools involved in cellular signalling, and to understand better the basis for intracellular [Ca2+]i oscillations. We also intend to develop a technique for real-time tracking of the [Ca2+]i in the lumen of intracellular stores. This work will involve a combinationof CCD camera imaging of cells using cytoplasmic and compartmentalized fluorescent calcium dyes together with computer assisted deconvolution, as well as confocal microscopy, and subcellular fractionation.

工作摘要：钙信号传导 不可驱行的细胞通常涉及从Ca2+动员 第二信使肌醇的细胞内存储 1,4,5-三磷酸盐（IP3）。但是，尽管很明显 Ca2+的细胞内存储的座位是的一部分，要么是 源自内质网，这些网站的精确轨迹 钙存储及其与其他细胞结构的关系是 不清楚。另外，IP3与其受体的相互作用在 这些CA2+商店通常会导致Ca2+释放的复杂动力学 并重新汇总，导致再生出现 细胞质内Ca2+（[Ca2+] I）振荡。这些商店 Ca2+的重要性很重要，因为它们是 [Ca2+] i通过动作在许多不同的单元格中生成的信号 激素，神经递质和生长因子。变化 这些商店的集中度也启动了新的信号传导 控制凋亡，基因表达等过程的途径， 和电容性钙进入。该项目的研究重点 是识别和定位涉及的细胞内Ca2+池 蜂窝信号传导，并更好地理解 细胞内[Ca2+] I振荡。我们还打算开发 [Ca2+] i的实时跟踪技术的技术 细胞内存储。这项工作将涉及CCD的组合 使用细胞质和分室化对细胞进行摄像机成像 荧光钙染料与计算机辅助 反卷积，共聚焦显微镜和亚细胞 分馏。