Investigating the role of the conserved transcriptional regulator Apterous on muscle development

项目总结 迫切需要开发治疗肌肉萎缩的方法，因为肌肉萎缩会增加 患有肌肉营养不良、肌病、癌症、肾衰竭和肺部疾病的患者。茎 细胞疗法需要产生特定大小和形状的肌肉来取代受损的肌肉的能力 肌肉，例如，圆眼或细长的肢体。透彻地了解肌肉是如何 特定的特性开发将允许我们对肌肉细胞进行编程以采用特定的特性，这是关键的一步 开发这类治疗方法。这项拟议工作的长期目标是利用果蝇， 果蝇，以确定体细胞发育的细胞机制 肌肉具有不同的大小、形状、方向、神经、连接和基因表达模式。 在每块肌肉中，这些特性由不同的基因调节因子集编码，包括 保守基因无翅(AP)、中线(Mid)和肌节同源盒(MSH)。我们专注于一项 胚胎腹肌的亚群，表达AP、Mid和MSH。我们已经发现AP的表达 在其正常模式之外，它会戏剧性地扰乱肌肉结构，导致肌肉位置和 失去肌肉附着物。肌肉-肌腱连接的失败阻止了这些突变胚胎 产生从蛋壳中孵化所需的力量并导致死亡。我们的假设是美联社直接 通过以下方式调节与肌肉定向和直接与间接肌肉连接选择有关的基因 在包括Mid和MSH在内的基因调控网络中发挥作用。我们将通过三个目标来检验这一假设： 1)检测AP突变背景中肌肉引导和依恋因子的位置和水平；2) 确定AP调控的基因；3)确定AP、MSH和Mid之间的相互作用 肌肉发育。这项拟议的工作将基因表达分析与细胞生物学、显微镜 以及遗传学，以了解肌肉连接是如何建立的，并确定有多少基因是AP的靶点 监管。综上所述，这项提案中描述的实验将决定肌肉特性 类似的形状，方向和依附类型被指定，这将有助于我们对脊椎动物的理解 人类肌肉发育的直系物，并导致治疗方法的发展。

PROJECT SUMMARY There is a critical need to develop treatments for muscle wasting, which increases morbidity and mortality of patients suffering from muscular dystrophies, myopathies, cancer, kidney failure and pulmonary disease. Stem cell therapies require the ability to generate muscles of particular sizes and shapes to replace damaged muscles in, for example, the round eye or the elongated limb. A thorough understanding of how muscles with specific properties develop will allow us to "program" muscle cells to adopt specific properties, a key step towards developing these kinds of treatments. The long term goal of the proposed work is to use the fruit fly, Drosophila melanogaster, to determine the cellular mechanisms underlying the development of somatic muscles with distinct sizes, shapes, orientations, innervations, attachments and gene expression patterns. Within each muscle these properties are encoded by distinct sets of gene regulatory factors, including the conserved genes Apterous (Ap), Midline (Mid), and Muscle-segment homebox (Msh). We are focused on a subset of embryonic abdominal muscles that express Ap, Mid and Msh. We have found that expression of Ap outside its normal pattern dramatically disrupts the musculature, leading to changes in muscle positioning and loss of muscle attachments. The failure of the muscle-tendon connection prevents these mutant embryos from generating the force needed to hatch from its eggshell and leads to death. Our hypothesis is that Ap directly regulates genes involved in muscle orientation and the selection of direct versus indirect muscle attachment by functioning in a network of gene regulators including Mid and Msh. We will test this hypothesis with three Aims: 1) examining the location and level of muscle guidance and attachment factors in Ap mutant backgrounds; 2) identifying the genes regulated by Ap; and 3) determining the interactions between Ap, Msh and Mid during muscle development. The proposed work combines gene expression analyses with cell biology, microscopy and genetics to learn how muscle connections are made and determine how many genes are targets of Ap regulation. Taken together, the experiments described in this proposal will determine how muscle properties like shape, orientation and attachment type are specified, which will inform our understanding of the vertebrate orthologs of muscle development in humans and lead to the development of therapies.