Molecular Mechanism and Functional Role of SOCE in Skeletal Muscle

SOCE在骨骼肌中的分子机制和功能作用

基本信息

批准号：
9906164
负责人：
Robert T Dirksen
金额：
$ 39.57万
依托单位：
UNIVERSITY OF ROCHESTER
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-08-01 至 2022-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9906164
关键词：
Adult Aging Area Biological Biology Biophysics Calcium Collaborations Complex Coupled Coupling Data Dependence Development Disease Dystrophin Electron Microscopy Exercise Exhibits Fatigue Female Fiber Functional disorder Gender Genes Growth Growth and Development function Heat Stress Disorders Immunologic Deficiency Syndromes Immunoprecipitation Knock-out Knockout Mice Lead Location Maintenance Mass Spectrum Analysis Membrane Molecular Mus Muscle Muscle Contraction Muscle Development Muscle Fatigue Muscle Fibers Muscle Weakness Muscle function Muscular Dystrophies Mutation Myopathy Nuclear Outcome Pathway interactions Patients Performance Permeability Pharmacology Phenotype Physiological Play Property Proteins Proteome Proteomics Publishing Recovery Regulation Reporting Research Resistance Role STIM1 gene Sarcoplasmic Reticulum Severe Combined Immunodeficiency Severities Signal Pathway Signal Transduction Skeletal Muscle Specificity Surface Tamoxifen Testing Time Triad Acrylic Resin Tubular Aggregate Myopathies base delta Sarcoglycan electron tomography extracellular frontier gain of function mutation gender difference immunocytochemistry in vivo insight loss of function mutation male mouse model multiple reaction monitoring muscular structure novel performance site postnatal public health relevance sarcopenia sensor skeletal muscle growth tool

项目摘要

DESCRIPTION (provided by applicant): Being first reported in 2001, store-operated Ca2+ entry (SOCE) is a relatively new phenomenon in skeletal muscle. SOCE is coordinated by coupling between two proteins: STIM1 calcium sensors in the sarcoplasmic reticulum (SR) and Ca2+-permeable Orai1 channels in the transverse tubule (TT) membrane. SOCE enhances muscle growth/development, limits fatigue, and promotes fatigue-resistant type I fiber specification. On the other hand, SOCE dysfunction contributes to muscle weakness/fatigue in aging, exacerbates muscular dystrophy, and mutations in STIM1 and Orai1 genes result in debilitating myopathies. The picture that emerges is that tight regulation of STIM1/Orai1-dependent SOCE activity is critical for optimal muscle performance such that increases or decreases in SOCE activity can lead to muscle fatigue, sarcopenia, and myopathy. While SOCE activity clearly impacts muscle performance, sites of STIM1-Orai1 coupling in muscle remain unclear. For this renewal, we developed inducible, muscle-specific Orai1 and STIM1 KO mice to determine the mechanism by which STIM1-Orai1 coupling limits fatigue. We provide exciting evidence that fatiguing exercise drives the formation of heretofore undescribed junctions between the TT and SR where STIM1-Orai1 coupling occurs, which we refer to as "Ca2+ entry units" (CEUs). CEUs are connected to, but distinct from, the triad or Ca2+ release unit. We provide preliminary data that Orai1 has a stronger impact on muscle fiber contractile function in female compared to male mice. We also provide preliminary collaborative immunoprecipitation and mass spectroscopy feasibility data for characterizing the STIM1 proteome before and after CEU formation. We will use these research tools, approaches, discoveries, and collaborations to advance understanding of the molecular determinants, subcellular location, and functional role of SOCE in skeletal muscle. Based on our published and preliminary data, we hypothesize that fatiguing exercise triggers formation of junctional extensions of the triad containing activated STIM1-Orai1 complexes that coordinate SOCE to enhance SR calcium refilling, limit muscle fatigue, and over the long-term, promote NFATc1 nuclear localization and type I fiber specification. We also hypothesize that fatigue-induced CEU formation in muscle involves a complex coordination of multiple protein components (including Bin1, STIM1, and Orai1). We propose to test these hypotheses according to the following two Specific Aims. Aim 1 will characterize the role of Orai1 in muscle fatigue and Type I fiber specification. Aim 2 will identif the subcellular location, molecular components, and stability of newly identified CEUs in adult skeletal muscle and determine the dependence of CEU formation and disassembly on the development of and recovery from fatigue. This project will: 1) provide novel mechanistic insights into the physiological role and subcellular location of SOCE in muscle, 2) use targeted and non-biased discovery approaches to identify and validate proteins involved CEU formation, and 3) determine the impact of gender on Orai1-dependent SOCE function, fatigue, fiber type specification, and CEU formation.

描述（由申请人提供）：2001 年首次报道，钙池操纵的 Ca2+ 进入（SOCE）是骨骼肌中相对较新的现象，SOCE 通过两种蛋白质之间的耦合进行协调：肌浆网（SR）中的 STIM1 钙传感器和肌浆网中的 STIM1 钙传感器。横管 (TT) 膜中的 Ca2+ 渗透性 Orai1 通道可增强肌肉生长/发育、限制疲劳并促进抗疲劳型。另一方面，SOCE 功能障碍会导致衰老时的肌肉无力/疲劳，使肌肉营养不良恶化，而 STIM1 和 Orai1 基因的突变会导致衰弱性肌病。由此可见，STIM1/Orai1 依赖性的严格调节。 SOCE 活动对于最佳肌肉表现至关重要，因此 SOCE 活动的增加或减少可能导致肌肉疲劳、肌肉减少症和肌病，而 SOCE 活动明显影响肌肉表现、部位。肌肉中的 STIM1-Orai1 偶联仍不清楚，我们开发了可诱导的肌肉特异性 Orai1 和 STIM1 KO 小鼠，以确定 STIM1-Orai1 偶联限制疲劳的机制。 TT 和 SR 之间未描述的连接处发生 STIM1-Orai1 耦合，我们将其称为“Ca2+ 输入单元”(CEU)。但与三联体或 Ca2+ 释放单元不同，我们提供了初步数据，表明与雄性小鼠相比，Orai1 对雌性小鼠的肌纤维收缩功能具有更强的影响。我们还提供了初步的协作免疫沉淀和质谱可行性数据，用于表征 STIM1 蛋白质组。在 CEU 形成后，我们将根据我们已发表的研究成果，利用这些研究工具、方法、发现和合作来加深对 SOCE 在骨骼肌中的分子决定因素、亚细胞定位和功能作用的理解。根据初步数据，我们发现疲劳运动会触发含有激活的 STIM1-Orai1 复合物的三联体连接延伸的形成，该复合物协调 SOCE 来增强 SR 钙再填充，限制肌肉疲劳，并从长远来看，促进 NFATc1 核定位和 I 型纤维我们还认为，疲劳诱导的肌肉中 CEU 的形成涉及多种蛋白质成分（包括 Bin1、STIM1 和 Orai1）的复杂协调，我们建议根据以下假设来检验这些假设。两个具体目标。目标 1 将描述 Orai1 在肌肉疲劳中的作用，目标 2 将确定成人骨骼肌中新鉴定的 CEU 的亚细胞位置、分子成分和稳定性，并确定 CEU 形成和的依赖性。该项目将：1）为肌肉中 SOCE 的生理作用和亚细胞位置提供新的机制见解，2）有针对性地使用和恢复。无偏见的发现方法来识别和验证涉及 CEU 形成的蛋白质，3) 确定性别对 Orai1 依赖性 SOCE 功能、疲劳、纤维类型规范和 CEU 形成的影响。

项目成果

期刊论文数量（34）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Pre-assembled Ca2+ entry units and constitutively active Ca2+ entry in skeletal muscle of calsequestrin-1 knockout mice.

DOI：
10.1085/jgp.202012617
发表时间：
2020-10-05
期刊：
The Journal of general physiology
影响因子：
0
作者：
Michelucci A;Boncompagni S;Pietrangelo L;Takano T;Protasi F;Dirksen RT
通讯作者：
Dirksen RT

Role of Mitofusin-2 in mitochondrial localization and calcium uptake in skeletal muscle.

DOI：
10.1016/j.ceca.2014.11.002
发表时间：
2015-01
期刊：
Cell calcium
影响因子：
4
作者：
Ainbinder A;Boncompagni S;Protasi F;Dirksen RT
通讯作者：
Dirksen RT

Addendum: Exercise-dependent formation of new junctions that promote STIM1-Orai1 assembly in skeletal muscle.

DOI：
10.1038/s41598-018-33063-0
发表时间：
2018-11-27
期刊：
Scientific reports
影响因子：
4.6
作者：
Boncompagni S;Michelucci A;Pietrangelo L;Dirksen RT;Protasi F
通讯作者：
Protasi F

Substrate-dependent and cyclophilin D-independent regulation of mitochondrial flashes in skeletal and cardiac muscle.

DOI：
10.1016/j.abb.2019.03.003
发表时间：
2019-04
期刊：
Archives of biochemistry and biophysics
影响因子：
3.9
作者：
L. Wei-LaPierre;Alina Ainbinder;Kevin M. Tylock;R. Dirksen
通讯作者：
L. Wei-LaPierre;Alina Ainbinder;Kevin M. Tylock;R. Dirksen

Antioxidants protect calsequestrin-1 knockout mice from halothane- and heat-induced sudden death.

DOI：
10.1097/aln.0000000000000748
发表时间：
2015-09
期刊：
Anesthesiology
影响因子：
8.8
作者：
Michelucci A;Paolini C;Canato M;Wei-Lapierre L;Pietrangelo L;De Marco A;Reggiani C;Dirksen RT;Protasi F
通讯作者：
Protasi F