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Mechanisms and Function of Myonuclear Positioning

肌核定位的机制和功能

基本信息

批准号：
10557796
负责人：
MARY K BAYLIES
金额：
$ 60.82万
依托单位：
SLOAN-KETTERING INST CAN RESEARCH
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-01 至 2026-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10557796
关键词：
3-Dimensional Adaptor Signaling Protein Address Architecture Area Biochemical Biological Biological Assay Biological Markers Biology CRISPR/Cas technology Cell Nucleus Cell physiology Cells Centronuclear myopathy Coculture Techniques Communication Cytoskeleton Data Development Disease Dissection Drosophila genus Drug usage Dynein ATPase Electrophysiology (science)Etiology Experimental Genetics Funding Genes Genetic Goals Growth Health Homeostasis Homologous Gene Human Human Cell Line Integrins Kinesin Knowledge Learning Link Literature MAPK8 gene Metabolic Metabolism Methodology Microtubule-Associated Proteins Microtubules Mitochondria Modeling Motor Motor Neurons Movement Muscle Muscle Cells Muscle Development Muscle Fibers Muscle function Myopathy Myotonic Dystrophy Neuromuscular Junction Normal Cell Nuclear Organelles Phenotype Physiological Positioning Attribute Process Proteins Publishing Research Role Sarcomeres Shapes Signal Pathway Signal Transduction Signaling Protein Striated Muscles System Tendon structure Testing Time Translating Work cell type fly genetic approach genetic manipulation genetic resource human disease imaging approach in vivo insight insulin signaling muscle physiology mutant neuromuscular neuromuscular activity ninein novel strategies proteostasis public health relevance regenerative therapeutic target three dimensional cell culture transcriptome sequencing

项目摘要

PROJECT SUMMARY/ABSTRACT Our long-term goal is to significantly impact the fundamental knowledge of muscle biology and provide new approaches for disease treatment. Striated muscle fibers are large multinucleated cells and possess a highly organized cytoarchitecture containing organelles positioned for optimal muscle function. This positioning is particularly evident in the placement of myonuclei, which reside above the sarcomere at the periphery of the myofiber and are positioned to maximize their internuclear distance. Our objective is the identification of mechanisms responsible for myonuclear movement and positioning. Centrally located myonuclei have been used for decades as a hallmark of muscle disease. However, much remains to be learned about the mechanisms that control myonuclear movement normally and the contribution of aberrant myonuclear position to the etiology and/or progression of muscle disease. Building on our published results over funding period (e.g. Metzger et al., 2012; Folker et al., 2012, 2014; Schulman et al., 2013, 2014; Azevedo et al., 2016; Manhart et al., 2018, 2020; Rosen et al., 2019), our specific aims are to first address mechanistically how tendon and motoneurons signal to the myofiber to fine-tune myonuclear positioning. We identified two signaling pathways at the myotendinous junction that regulate nuclear positioning. Likewise, we will dissect the contribution of the motoneuron to nuclear placement. Secondly, we will examine why muscles fail to function optimally when myonuclei are mispositioned. Muscle physiology will be assayed through testing mitochondrial function via quantification of ATP and ROS levels and by testing neuromuscular communication via electrophysiological approaches. We will also investigate the input to muscle function of specific metabolic and signaling proteins that we identified are misregulated as a result of mispositioned myonuclei. Thirdly, we will push forward our dissection of the myonuclear positioning mechanisms from fly to the human system. We will employ human 3D muscle cultures that are co-cultured with motoneurons to define and then perturb myonuclear movement and positioning as the myofibers develop. Our methodologies take advantage of cutting edge, in vivo time lapse imaging approaches that we have developed in Drosophila and will also apply to human 3D cultures to follow myonuclear movement and cytoskeletal dynamics. We will employ the genetic resources available in Drosophila and human cultures to manipulate genes, processes, and cell types for our analyses. These genetic experiments will be supported by biochemical and cell biological approaches. Together the work outlined in this proposal will shed new light on this little understood, but important area of muscle biology. The results of this research will permit us to highlight genes and mechanisms that are candidates for changes associated with different human muscle diseases.

项目总结/摘要我们的长期目标是显著影响肌肉生物学的基础知识，并提供新的疾病治疗的方法。横纹肌纤维是大的多核细胞，具有高度的有组织的细胞结构，包含定位于最佳肌肉功能的细胞器。该定位在肌核的放置中特别明显，肌核位于肌节周围的肌节上方，肌纤维和定位，以最大限度地提高其核间距离。我们的目标是识别负责肌肉运动和定位的机制。位于中央的肌细胞核几十年来一直被用作肌肉疾病的标志。然而，关于这些机制，控制正常肌运动的神经系统，以及异常肌位置对病因的贡献和/或肌肉疾病的进展。基于我们在资助期内发表的结果（例如Metzger等人， 2012; Folker等人，2012，2014; Schulman等人，2013，2014; Azevedo等人，2016; Manhart等人，2018年，2020年; 罗森等人，我们的具体目标是首先从机制上解决肌腱和运动神经元如何发出信号来微调肌纤维的位置我们在肌腱中发现了两条信号通路，调节核定位的结。同样，我们也将分析运动神经元对核神经元的作用。安置其次，我们将研究为什么肌肉不能发挥最佳功能时，肌核错位。将通过ATP和ROS定量检测线粒体功能来分析肌肉生理学水平和通过电生理方法测试神经肌肉通信。我们亦会研究我们确定的特定代谢和信号蛋白对肌肉功能的输入是错误调节的，结果是肌核错位。第三，我们将推进对肌核定位的解剖从苍蝇到人类系统的机制。我们将采用与人3D肌肉共培养的人3D肌肉培养物，运动神经元，以确定，然后扰乱肌纤维的发展，肌运动和定位。我们这些方法利用了我们开发的尖端的体内时间推移成像方法，在果蝇中，也将应用于人类3D培养，以跟踪果蝇运动和细胞骨架动力学我们将利用果蝇和人类文化中的遗传资源来操纵基因，过程和细胞类型进行分析。这些遗传实验将得到生物化学和细胞生物学的支持。生物学方法。本提案中概述的工作将共同揭示这一鲜为人知的问题，但是肌肉生物学的重要领域。这项研究的结果将使我们能够突出基因，这些机制是与不同人类肌肉疾病相关的变化的候选者。