Skeletal Muscle. Ca Release Control Inside the Sarcoplasmic Reticulum.

骨骼肌。

• 批准号: 8067962
• 负责人: Eduardo Rios
• 金额: $ 30.7万
• 依托单位: RUSH UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-07-01 至 2014-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8067962
• 关键词: Address; Adult; Affect; Age; Aging; Amphibia; Animals; Area; Binding; Binding Proteins; Biochemical; Biochemistry; Biosensor; Body Weight; Buffers; Calcium; Calcium Signaling; Calcium-Binding Proteins; Calsequestrin; Cell physiology; Cells; DNA; Defect; Disease; Employee Strikes; Endowment; Evaluation; Evolution; Exercise; Exposure to; Failure; Fatigue; Fiber; Figs - dietary; Frequencies; Functional disorder; Gene Expression; Gene Silencing; Gifts; Goals; Health; Histidine; Hour; Hybrids; Image; Immune; Immune system; Infection; Knock-out; Language; Lead; Life; Lymphocyte; Malignant hyperpyrexia due to anesthesia; Mammals; Measurement; Measures; Mediating; Membrane; Metabolism; Methods; Mitochondria; Modification; Monitor; Motion; Movement; Mus; Muscle; Muscle Cells; Muscle Fatigue; Muscle Proteins; Muscular Dystrophies; Myopathy; Optics; Organ; Pathway interactions; Pattern; Performance; Permeability; Phenotype; Play; Predisposition; Property; Proteins; Protocols documentation; Regulation; Relaxation; Research; Rest; Role; STIM1 gene; Sarcoplasmic Reticulum; Secure; Signal Transduction; Skeletal Muscle; Stimulus; Strenuous Exercise; System; Techniques; Testing; Therapeutic; Time; Tissues; Transfection; Universities; Wisconsin; Work; Yang; base; calreticulin; chromophore; circumsporozoite protein; combat; controlled release; density; design; improved; indexing; inhibitor/antagonist; insight; interest; killings; knock-down; millisecond; mouse junctate protein; muscle aging; novel; novel strategies; overexpression; physical conditioning; prevent; protein distribution; research study; sensor; tool; transcription factor; triadin; voltage clamp

DESCRIPTION (provided by applicant): Ca2+ signaling is a universal language used by cells to react and change. In skeletal muscle its patterns of interest cover multiple time scales: milliseconds -Ca2+ movements that determine contraction and relaxation-; seconds to minutes -when sustained activity may lead to myogenic fatigue- and hours to weeks -patterns that cause changes in gene expression and long-term adaptation-. This study is about inside its cellular store; its quantity, and its concentration, [Ca2+]SR, which conditions Ca2+ signals in every time scale. We ask (1) whether and how [Ca2+]SR controls Ca2+ release from the store, and (2) whether and how calsequestrin and triadin, two strategically located SR proteins, contribute to this control. A technical task, which we call "aim 0", is to image and measure [Ca2+]SR. This was accomplished in the current period and will continue in the next, using novel biosensors -molecules made by the cells themselves- and new hybrid monitors, consisting of high performance small synthetic sensors placed into cells manipulated to make special bio-anchors. To answer questions 1 and 2, we will respectively manipulate [Ca2+]SR while we measure it (aim 1) and force cells to change their endowment of calsequestrin and triadin (aim 2). These goals are now feasible in living animals thanks to a DNA transfection method that works with every protein and can be used also to prevent their synthesis. We propose that [Ca2+]SR -which decays when muscles fatigue- is sustained by SOCE, a universal Ca2+ entry pathway, crucial for mobilizing transcription factors that control gene expression. Using SOCE measures developed in the first period, we propose as aim 3 to define the role of newly discovered molecules of SOCE in the control of [Ca2+]SR. These molecules could be bulwarks against fatigue, and provide powerful tools for experimental alterations of [Ca2+]SR in iterative approaches to the main questions. Ca2+ signals deteriorate in disease, fatigue and aging. Fast Ca2+ signals fail in diseases like hypo-PP, MH susceptibility and central core and minicore, as well as in ageing muscle. Mid-range signaling is affected in fatigue and in an MH-like phenotype of mice lacking calsequestrin. Diseases of long term Ca2+ signals, which show striking parallels in muscle and the immune system, include SCID, a familial immune defect that combines loss of SOCE in lymphocytes and a myogenic myopathy. Our work will advance understanding of these deficits by evaluating roles of specific molecules and their interactions. While only fatigue will be specifically addressed in the present project, questions on the relationships among deficits of function, the intricate pathophysiology and the rational design of therapeutic corrections will be addressed better as we understand what controls stored calcium, and what the stored calcium controls. PUBLIC HEALTH RELEVANCE: This project and our lab's work deal with movements of calcium inside muscle. So-called "calcium signaling" is a universal language used by cells to react and change. In skeletal muscle these signals make the difference between rest and motion, thus influencing multiple aspects of our life, including physical conditioning, metabolism, and even body weight. Because calcium is used for many tasks, including killing cells, the signals are dangerous and must be kept under control. In this project we ask what controls the movements of calcium, in particular focusing on roles of calcium storage areas inside the cell. We also ask how the intra-store calcium determines how much will move, or when the movement will cease. Therefore, our work should help understand why signals fail, or get out of hand. This occurs in diseases, including muscular dystrophies, malignant hyperthermia, and others. Signals also decay during fatigue -their decay actually causing fatigue-- and in old age. In muscle fatigue, the stores are depleted. We propose that two newly discovered molecules are important factors that delay this depletion. These molecules are known to play roles in organs and cells other than muscle, including cells that combat infections. As a result there will be multiple repercussions to their defects or failures. It can be anticipated, therefore, that our research will have consequences outside its field of muscle and exercise.

描述（由申请人提供）：Ca2+信号传导是细胞用于反应和变化的通用语言。在骨骼肌中，其感兴趣的模式涵盖多个时间尺度：毫秒-决定收缩和松弛的Ca2+运动；几秒到几分钟（持续活动可能导致肌源性疲劳）和几小时到几周（导致基因表达和长期适应变化的模式）。这项研究是关于其细胞存储的内部；它的数量和浓度，[Ca2+]SR，调节每个时间尺度的 Ca2+ 信号。我们询问 (1) [Ca2+]SR 是否以及如何控制 Ca2+ 从储存库中释放，以及 (2) 钙螯合蛋白和三联蛋白（两种战略定位的 SR 蛋白）是否以及如何有助于这种控制。我们称之为“目标 0”的技术任务是成像和测量 [Ca2+]SR。这项工作已在当前阶段完成，并将在下阶段继续进行，使用新型生物传感器（由细胞本身产生的分子）和新型混合监测器，其中包括放置在细胞中的高性能小型合成传感器，经操纵以制造特殊的生物锚。为了回答问题 1 和 2，我们将在测量时分别操纵 [Ca2+]SR（目标 1）并迫使细胞改变其钙螯合蛋白和三联蛋白的禀赋（目标 2）。得益于 DNA 转染方法，这些目标现在可以在活体动物中实现，该方法适用于每种蛋白质，并且还可以用于阻止其合成。我们提出，[Ca2+]SR（当肌肉疲劳时会衰减）由 SOCE 维持，SOCE 是一种通用的 Ca2+ 进入途径，对于动员控制基因表达的转录因子至关重要。使用第一阶段开发的 SOCE 措施，我们提出目标 3 来定义新发现的 SOCE 分子在 [Ca2+]SR 控制中的作用。这些分子可以成为对抗疲劳的堡垒，并为解决主要问题的迭代方法中 [Ca2+]SR 的实验改变提供强大的工具。 Ca2+ 信号会因疾病、疲劳和衰老而恶化。快速 Ca2+ 信号在低 PP、MH 易感性、中央核心和小核心以及老化肌肉等疾病中失效。中程信号传导在疲劳和缺乏钙螯合蛋白的 MH 样表型小鼠中受到影响。长期 Ca2+ 信号疾病在肌肉和免疫系统中表现出惊人的相似之处，包括 SCID，这是一种家族性免疫缺陷，结合了淋巴细胞 SOCE 的缺失和肌源性肌病。我们的工作将通过评估特定分子的作用及其相互作用来增进对这些缺陷的理解。虽然本项目将专门解决疲劳问题，但当我们了解什么控制储存的钙以及储存的钙控制什么时，有关功能缺陷、复杂的病理生理学和治疗校正的合理设计之间关系的问题将得到更好的解决。公共健康相关性：该项目和我们实验室的工作涉及肌肉内钙的运动。所谓的“钙信号传导”是细胞用来反应和变化的通用语言。在骨骼肌中，这些信号决定休息和运动，从而影响我们生活的多个方面，包括身体状况、新陈代谢，甚至体重。由于钙用于许多任务，包括杀死细胞，因此这些信号是危险的，必须受到控制。在这个项目中，我们询问是什么控制着钙的运动，特别关注细胞内钙存储区域的作用。我们还询问店内钙如何确定移动的程度，或者移动何时停止。因此，我们的工作应该有助于理解信号失败或失控的原因。这种情况发生在肌肉萎缩症、恶性高热等疾病中。信号也会在疲劳期间和老年时衰减（它们的衰减实际上会导致疲劳）。在肌肉疲劳时，储存会耗尽。我们认为两个新发现的分子是延迟这种消耗的重要因素。已知这些分子在肌肉以外的器官和细胞中发挥作用，包括抵抗感染的细胞。因此，它们的缺陷或失败将会产生多重影响。因此，可以预见，我们的研究将在肌肉和运动领域之外产生影响。