The mechanisms and functions of nuclear spacing in muscle development

核间距在肌肉发育中的机制和功能

• 批准号: 10733794
• 负责人: Eric S Folker
• 金额: $ 31.3万
• 依托单位: BOSTON COLLEGE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10733794
• 关键词: Ablation; Address; Animals; Behavior; Binding; Binding Proteins; Biological; Biological Assay; Cell Nucleus; Cell physiology; Cells; Chimeric Proteins; Complex; Cytoskeleton; Data; Development; Disease; Drosophila melanogaster; Duchenne muscular dystrophy; Dystrophin; Embryo; Eukaryota; Genes; Genetic; Goals; Golgi Apparatus; Growth; Homeostasis; Imaging Device; Impairment; Individual; Intuition; Investigation; Kinesin; Knowledge; Larva; Lasers; Maintenance; Manuscripts; Measures; Membrane; Methods; Microscopy; Microtubule-Organizing Center; Microtubules; Modeling; Molecular; Movement; Mus; Muscle; Muscle Cells; Muscle Development; Muscle Fibers; Muscle function; Mutate; Myopathy; Nature; Nuclear; Outcome; Pathogenesis; Population; Positioning Attribute; Prevalence; Process; Property; Proteins; Publishing; Regulation; Research; Role; Sarcolemma; Series; Slide; Testing; Work; cellular development; density; experimental study; genetic approach; in vivo; insight; mutant

Summary Nuclear movement is evolutionarily conserved throughout eukaryotes, a testament to the fundamentally important nature of this process. The importance of this process is most evident in syncytial muscle cells, which move their nuclei to maximize the distance between each nucleus and its neighbors, a process thought to establish a series of independent myodomains. Furthermore, mispositioned nuclei in muscle cells are prevalent in muscle disease. In recent years, dozens of proteins have been implicated in the movement and spacing of nuclei, but their mechanisms of action, and the consequences of nuclear spacing, remain elusive. The goal of the proposed research is to fill this critical gap in knowledge. Specifically, we will define the features of the microtubule cytoskeleton that are critical for both moving nuclei to establish the spacing and anchor nuclei to maintain the spacing. Only recently have the molecular, physical, and imaging tools emerged to make it possible to address these mechanistic questions in vivo. In the proposed research we will generate and test mutant proteins that target the microtubule specific functions of pleitropic proteins to isolate the specific contribution of the microtubule interactions. We will then apply our recently developed analyses of nuclear movement and microtubule organization combined with classical genetic approaches to determine the genetic and molecular mechanisms by which these factors move and anchor nuclei during muscle development. Within this work we will test whether two emergent properties of the microtubule cytoskeleton, branching and sliding contribute to nuclear spacing in muscle. Finally, we will directly tie these data to disease pathogenesis by determining how dystrophin, the gene mutated in the most common and debilitating muscle disease contributes to these processes. Successful completion of these aims will provide the first mechanistic understanding of nuclear movement in an in-tact developing muscle cell making it possible to manipulate nuclear spacing to understand how the position of the nucleus impacts its function.

摘要 在真核生物中，核运动在进化上是保守的，这证明了 这是这一进程的重要性质。这一过程的重要性在合体肌细胞中最为明显， 它们移动原子核以最大化每个原子核与其邻居之间的距离，这是一种过程思想 建立一系列独立的肌结构域。此外，肌肉细胞中错位的细胞核是 流行于肌肉疾病。近年来，数十种蛋白质参与了这一运动和 原子核之间的距离很小，但它们的作用机制以及核间距的后果仍然难以捉摸。 拟议研究的目标是填补这一关键的知识空白。具体地说，我们将定义 微管细胞骨架的特征，它对移动的核建立间距和 锚定原子核以保持间距。直到最近，分子、物理和成像工具才出现 使在活体内解决这些机械问题成为可能。在建议的研究中，我们将生成 并测试以微管为靶点的突变蛋白，以分离多发性蛋白的特定功能 微管相互作用的具体贡献。然后，我们将应用我们最近开发的分析 核运动和微管组织结合经典遗传学方法确定 这些因子在肌肉过程中移动和锚定细胞核的遗传和分子机制 发展。在这项工作中，我们将测试微管细胞骨架的两个新特性， 分支和滑动有助于肌肉中的核间距。最后，我们将把这些数据直接与疾病联系起来 通过确定dystrophin基因如何在最常见和最虚弱的肌肉中突变来确定其发病机制 疾病促成了这些过程。这些目标的成功实现将提供第一个机制 理解发育中的肌肉细胞中的核运动使操控成为可能 原子核间距，以了解原子核的位置如何影响其功能。