Molecular Mechanisms of Myoblast Fusion

描述（申请人提供）：骨骼肌是一种独特的器官，由多核肌纤维组成，每一个肌纤维都是数百甚至数千个成肌细胞融合的产物。成肌细胞融合不仅对骨骼肌发育非常重要，而且对基于卫星细胞的肌肉再生也至关重要。尽管几十年来进行了大量的研究，但人类成肌细胞融合的机制仍然知之甚少。最近在果蝇中的研究已经开始揭示关于成肌细胞融合的分子和细胞机制的前所未有的细节。果蝇和哺乳动物肌发生之间惊人的进化保守性使得果蝇成为研究体内成肌细胞融合的特别相关的系统。本实验室最近的研究揭示了成肌细胞融合的一种新的细胞机制。我们发现，成肌细胞融合介导的细胞类型特异性，F-肌动蛋白富集的podosome样结构（PLS），它侵入对面的融合伴侣与多个膨胀的手指，导致融合孔的形成。为了进一步表征侵入性PLS的调节，介导成肌细胞融合的关键细胞结构，我们建议研究两个新基因DPak 3和发动蛋白的分子和细胞功能，这两个基因以前都与人类疾病有关。我们将使用多方面的方法，包括遗传学，细胞生物学和生物化学，研究DPak 3和动力蛋白调节果蝇成肌细胞融合PLS内肌动蛋白聚合动力学的机制。我们还将研究它们的哺乳动物直系同源物在小鼠C2 C12成肌细胞融合中的功能。鉴于果蝇和哺乳动物之间成肌细胞融合的分子和细胞保守性，使用更简单和遗传上易于处理的果蝇系统进行的机制研究将导致对人类肌肉生物学在健康和疾病方面的重要见解，并最终为开发更有效的治疗方法提供基础。

DESCRIPTION (provided by applicant): Skeletal muscle is a unique organ that is composed of multinucleate muscle fibers, each of which is the product of fusion of hundreds or even thousands of myoblasts. Myoblast fusion is not only important for skeletal muscle development, but also critical for satellite cell-based muscle regeneration. Despite a large body of studies ove several decades, the mechanisms underlying myoblast fusion in humans remain poorly understood. Recent studies in the fruit fly Drosophila have begun to reveal unprecedented details about the molecular and cellular mechanisms of myoblast fusion. The striking evolutionary conservation between fly and mammalian myogenesis makes Drosophila a particularly relevant system to study myoblast fusion in vivo. Recent studies from our lab have uncovered a novel cellular mechanism underlying myoblast fusion. We show that myoblast fusion is mediated by a cell type-specific, F-actin-enriched podosome-like structure (PLS), which invades the opposing fusion partner with multiple protrusive fingers leading to fusion pore formation. To further characterize the regulation of the invasive PLS, the key cellular structure mediating myoblast fusion, we propose to investigate the molecular and cellular functions of two new genes, DPak3 and dynamin, both have previously been implicated in human diseases. We will use a multifaceted approach, including genetics, cell biology and biochemistry, to study the mechanisms by which DPak3 and dynamin regulate actin polymerization dynamics within the PLS in Drosophila myoblast fusion. We will also investigate the function of their mammalian orthologs in the fusion of mouse C2C12 myoblasts. Given the molecular and cellular conservation of myoblast fusion between fly and mammals, the proposed mechanistic studies using the simpler and genetically tractable Drosophila system will lead to significant insights int human muscle biology in health and disease, and ultimately provide a basis for developing more efficient therapeutics against the life debilitating muscle degeneration diseases.