Skeletal Muscle. Ca Release Control Inside the Sarcoplasmic Reticulum.

骨骼肌。

• 批准号: 8268539
• 负责人: Eduardo Rios
• 金额: $ 30.71万
• 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/8268539
• 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+信号是细胞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- ca2 +运动决定收缩和松弛-；几秒到几分钟——持续的活动可能会导致肌肉疲劳——几小时到几周——这种模式会导致基因表达和长期适应的变化。这项研究是关于它的细胞储存；它的数量，它的浓度，[Ca2+]SR，它在每个时间尺度上控制Ca2+信号。我们询问(1)[Ca2+]SR是否以及如何控制Ca2+从储存中释放，以及(2)calsequestrin和triadin这两种战略性SR蛋白是否以及如何促进这种控制。一项技术任务，我们称之为“目标0”，是成像和测量[Ca2+]SR。这项工作已经在当前阶段完成，并将在接下来的阶段继续进行，使用新型生物传感器（由细胞自身产生的分子）和新型混合监测器，这些监测器由高性能的小型合成传感器组成，放置在被操纵的细胞中，以制造特殊的生物锚。为了回答问题1和2，我们将在测量[Ca2+]SR时分别操纵它（目标1），并迫使细胞改变钙sequestrin和triadin的禀赋（目标2）。这些目标现在在活体动物中是可行的，这要归功于一种DNA转染方法，这种方法可以与每种蛋白质一起工作，也可以用来阻止它们的合成。我们提出[Ca2+]SR -当肌肉疲劳时衰减-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。利用第一阶段开发的SOCE测量方法，我们提出了目标3，以确定新发现的SOCE分子在控制[Ca2+]SR中的作用。这些分子可能是抗疲劳的堡垒，并为[Ca2+]SR的实验改变提供了强大的工具，在迭代方法中解决主要问题。Ca2+信号在疾病、疲劳和衰老中恶化。快速Ca2+信号在低pp、MH易感性、中央核心和小核心以及衰老肌肉等疾病中失效。缺乏钙sequestrin的小鼠疲劳和mh样表型中，中程信号传导受到影响。长期Ca2+信号的疾病，在肌肉和免疫系统中显示出惊人的相似性，包括SCID，一种家族性免疫缺陷，结合淋巴细胞中SOCE的丧失和肌源性肌病。我们的工作将通过评估特定分子的作用及其相互作用来促进对这些缺陷的理解。虽然在目前的项目中只有疲劳将被具体解决，但随着我们了解是什么控制了钙的储存，又是什么控制了钙的储存，关于功能缺陷、复杂的病理生理学和治疗纠正的合理设计之间的关系的问题将得到更好的解决。公共卫生相关性：本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”是细胞用于反应和变化的通用语言。在骨骼肌中，这些信号决定了休息和运动的不同，从而影响我们生活的多个方面，包括身体调节、新陈代谢，甚至体重。因为钙被用于许多任务，包括杀死细胞，所以这些信号是危险的，必须加以控制。在这个项目中，我们询问是什么控制了钙的运动，特别关注细胞内钙储存区域的作用。我们也会问储存中的钙是如何决定有多少会移动，或者移动何时会停止。因此，我们的工作应该有助于理解信号失败或失控的原因。这发生在疾病中，包括肌肉萎缩症、恶性高热和其他疾病。信号在疲劳时也会衰减——它们的衰减实际上会导致疲劳——在老年时也会衰减。肌肉疲劳时，储存的能量会耗尽。我们认为，两种新发现的分子是延缓这种耗竭的重要因素。已知这些分子在肌肉以外的器官和细胞中发挥作用，包括对抗感染的细胞。因此，它们的缺陷或失败将产生多重影响。因此，可以预见的是，我们的研究将在肌肉和运动领域之外产生影响。