INTRACELLULAR CA2+ STORES AND THE IP3 RECEPTOR

细胞内 CA2 存储和 IP3 受体

基本信息

批准号：
6432425
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
项目类别：
财政年份：
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/6432425
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

项目摘要

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 of Ca2+ 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+] in the lumen of intracellular stores. This work will involve a combination of 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：非驱动细胞中的钙信号传导通常涉及第二信使，肌醇1,4,5-三磷酸（IP3）从细胞内存储中的Ca2+。然而，尽管很明显，Ca2+的细胞内存储的轨迹要么是内质网的一部分或源自内质网的一部分，但这些钙存储的精确基因座及其与其他细胞结构的关系尚不清楚。此外，IP3与其受体在这些CA2+存储上的相互作用通常会导致Ca2+释放和重新汇总的复杂动力学，从而导致再生细胞质内CA2+（[Ca2+] I）振荡的出现。这些CA2+的存储很重要，因为它们是通过激素，神经递质和生长因子的作用在许多不同细胞类型中产生的[Ca2+] I信号的初始来源。这些商店中Ca2+浓度的变化还启动了一种新的信号通路，该途径控制凋亡，基因表达和电容性钙进入等过程。该项目的研究重点是识别和定位与细胞信号传导有关的细胞内Ca2+池，并更好地理解细胞内[Ca2+] I振荡的基础。我们还打算开发一种用于在细胞内存储腔内[Ca2+]实时跟踪的技术。这项工作将涉及使用细胞质和分室化荧光钙染料与计算机辅助反卷积以及共聚焦显微镜以及亚细胞分级的CCD摄像机成像的组合。