Functional architecture of IP3-evoked local Ca2+ signals

IP3诱发的局部Ca2信号的功能架构

基本信息

批准号：
8214675
负责人：
JONATHAN S MARCHANT
金额：
$ 30.43万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-07-01 至 2014-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8214675
关键词：
Address Agonist Architecture Autoimmune Diseases Behavior Binding Binding Sites Biological Assay Cell physiology Cells Cellular biology Complement Congenital Heart Defects Coupled Cues Data Data Set Development Disease Embryo Embryonic Development Endoplasmic Reticulum Equilibrium Event Family Functional disorder Funding Gene Family Global Change Goals Health Heart Diseases ITPR1 gene Image In Vitro Individual Inositol Intracellular Membranes Ion Channel Kidney Diseases Kinetics Label Life Ligands Malignant Neoplasms Measurement Methods Microscope Molds Molecular NAADP Nerve Degeneration Neurodegenerative Disorders Outcome Pathway interactions Pattern Physiological Property Proteins Pump Reagent Regulation Resolution Role Signal Pathway Signal Transduction Site Stimulus Structure System Testing Time Tissues Translating Virus Diseases Work Xenopus cell type design egg flexibility fluorophore in vivo insight interest nervous system disorder novel oocyte maturation optical imaging receptor response single molecule spatiotemporal tool

项目摘要

DESCRIPTION (provided by applicant): Increases in cytoplasmic Ca2+ evoked by inositol(1,4,5)-trisphosphate receptors (IP3Rs) regulate many physiological events. Dysfunction of the IP3-stimulated Ca2+ release pathway is involved in neurodegenerative and neurological disease, as well as exocrine, autoimmune and kidney disorders, cardiac abnormalities and cancer. To appreciate how cellular functions are controlled by Ca2+ signals, and how pathological aberrations subvert the IP3R signaling pathway, we must understand how the distribution and properties (the `functional architecture') of IP3Rs control the spatiotemporal organization of cellular Ca2+ signals. Here, we will resolve the properties of IP3Rs at three distinct levels of organization from single molecules in vitro (Aim 1) to local, subcellular (Aim 2) and global, tissue-level IP3R architecture in live embryos (Aim 3). We will address: (1) How, for individual IP3R, does a single molecule of IP3 bind to the IP3R? (2) How is IP3R function modulated within the dynamic context of endoplasmic reticulum structures where they reside? (3) How does a changing global complement of Ca2+ channels and pumps at a regional level in a developing vertebrate embryo impacts the patterning of Ca2+ signals and cell function. To address these challenges, we have optimized: (i) a novel single molecule imaging approach competent to resolve the properties of individual IP3Rs (Aim 1), (ii) a dual confocal microscope to simultaneously resolve ER architecture and the functionality of IP3Rs (Aim 2) and (iii) designed tools and reagents to probe the distribution and role of key families of Ca2+ channels and pumps during vertebrate embryogenesis (Aim 3). The broad significance of this work is in understanding principles controlling ion channel dynamics and thereby the spatial kinetics of Ca2+ signals that control unitary, cellular and systems-level responses. Such data will aid our understanding of the role of ubiquitous Ca2+ signaling pathways in health and disease. PUBLIC HEALTH RELEVANCE: The goal of this project is to understand how particular cell signaling events are organized within cells by understanding first how proteins work at the unitary level in vitro and then appreciating how these proteins work when organized within cells and within developing embryos in vivo. The goal is to provide deeper understand of how intracellular Ca2+ channels behave and how pathological events subvert their function.

描述（由申请人提供）：肌醇（1,4,5）-三磷酸受体（IP3R）引起的细胞质Ca2+增加调节许多生理事件。 IP3 刺激的 Ca2+ 释放途径的功能障碍与神经退行性和神经系统疾病以及外分泌、自身免疫和肾脏疾病、心脏异常和癌症有关。为了了解 Ca2+ 信号如何控制细胞功能，以及病理畸变如何破坏 IP3R 信号通路，我们必须了解 IP3R 的分布和特性（“功能架构”）如何控制细胞 Ca2+ 信号的时空组织。在这里，我们将解析 IP3R 在三个不同组织层面的特性，从体外单分子（目标 1）到活胚胎中的局部、亚细胞（目标 2）和全局组织级 IP3R 结构（目标 3）。我们将讨论：（1）对于单个IP3R，单个IP3分子如何与IP3R结合？ (2) IP3R 功能如何在内质网结构所在的动态环境中进行调节？ (3) 发育中的脊椎动物胚胎在区域水平上不断变化的 Ca2+ 通道和泵的全局补充如何影响 Ca2+ 信号和细胞功能的模式。为了应对这些挑战，我们优化了：(i) 一种新颖的单分子成像方法，能够解析单个 IP3R 的特性（目标 1），(ii) 双共焦显微镜，可同时解析 ER 架构和 IP3R 的功能（目标2) 和 (iii) 设计了工具和试剂来探测脊椎动物胚胎发生过程中 Ca2+ 通道和泵关键家族的分布和作用（目标 3）。这项工作的广泛意义在于理解控制离子通道动力学的原理，从而理解控制单一、细胞和系统级响应的 Ca2+ 信号的空间动力学。这些数据将有助于我们了解普遍存在的 Ca2+ 信号通路在健康和疾病中的作用。公共健康相关性：该项目的目标是通过首先了解蛋白质如何在体外单一水平上发挥作用，然后了解这些蛋白质在细胞内和体内发育胚胎中组织时如何发挥作用，来了解特定的细胞信号传导事件如何在细胞内组织。。目标是更深入地了解细胞内 Ca2+ 通道的行为方式以及病理事件如何破坏其功能。

项目成果

期刊论文数量（10）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Molecular characterization of a novel cell surface ADP-ribosyl cyclase from the sea urchin.

DOI：
10.1016/j.cellsig.2008.09.005
发表时间：
2008-12
期刊：
CELLULAR SIGNALLING
影响因子：
4.8
作者：
Churaman, Dev;Boulware, Michael J.;Ramakrishnan, Latha;Geach, Timothy J.;Martin, Andrew C. R.;Vacquier, Victor D.;Marchant, Jonathan S.;Dale, Leslie;Patel, Sandip
通讯作者：
Patel, Sandip

Molecular characterization of a novel intracellular ADP-ribosyl cyclase.

DOI：
10.1371/journal.pone.0000797
发表时间：
2007-08-29
期刊：
PLOS ONE
影响因子：
3.7
作者：
Churamani, Dev;Boulware, Michael J.;Geach, Timothy J.;Martin, Andrew C. R.;Moy, Gary W.;Su, Yi-Hsien;Vacquier, Victor D.;Marchant, Jonathan S.;Dale, Leslie;Patel, Sandip
通讯作者：
Patel, Sandip

Tight junction targeting and intracellular trafficking of occludin in polarized epithelial cells.

极化上皮细胞中紧密连接靶向和occludin 的细胞内运输。

DOI：
10.1152/ajpcell.00309.2007
发表时间：
2007
期刊：
American journal of physiology. Cell physiology
影响因子：
0
作者：
Subramanian,VeedamaliS;Marchant,JonathanS;Ye,Dongmei;Ma,ThomasY;Said,HamidM
通讯作者：
Said,HamidM

The inositol 1,4,5-trisphosphate receptor (Itpr) gene family in Xenopus: identification of type 2 and type 3 inositol 1,4,5-trisphosphate receptor subtypes.

非洲爪蟾肌醇 1,4,5-三磷酸受体 (Itpr) 基因家族：2 型和 3 型肌醇 1,4,5-三磷酸受体亚型的鉴定。

DOI：
10.1042/bj20070101
发表时间：
2007
期刊：
The Biochemical journal
影响因子：
0
作者：
Zhang,Dan;Boulware,MichaelJ;Pendleton,MatthewR;Nogi,Taisaku;Marchant,JonathanS
通讯作者：
Marchant,JonathanS

Enhanced Ca2+ leak from ER Ca2+ stores induced by hepatitis C NS5A protein.