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

项目摘要 迫切需要开发针对肌肉萎缩的治疗方法，肌肉萎缩增加了肌肉萎缩症的发病率和死亡率。 患有肌肉萎缩症、肌病、癌症、肾衰竭和肺病的患者。干 细胞疗法需要产生特定大小和形状的肌肉的能力， 例如，圆形眼睛或细长肢体中的肌肉。彻底了解肌肉如何与 开发特定的特性将使我们能够"编程"肌肉细胞，使其具有特定的特性，这是关键的一步。 来发展这些治疗方法。这项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、神经支配、附件和基因表达模式。 在每块肌肉中，这些特性由不同的基因调控因子编码，包括 保守基因Apterous（Ap）、Midline（Mid）和Muscle-segment homebox（Msh）。我们专注于 表达Ap、Mid和Msh胚胎腹部肌肉的子集。我们已经发现， 在其正常模式之外，会显著破坏肌肉组织，导致肌肉定位的变化， 失去肌肉附着。肌肉-肌腱连接的失败阻止了这些突变胚胎 产生从蛋壳中孵化出来的力量并导致死亡。我们的假设是， 调节基因参与肌肉方向和选择直接与间接肌肉附着， 在包括Mid和Msh在内的基因调控网络中发挥作用。我们将以三个目标来检验这一假设： 1)检查Ap突变背景中肌肉引导和附着因子的位置和水平; 2） 鉴定由Ap调节的基因;和3）确定在细胞周期中Ap、Msh和Mid之间的相互作用。 肌肉发育拟议的工作结合了基因表达分析与细胞生物学，显微镜 和遗传学来了解肌肉连接是如何形成的，并确定有多少基因是Ap的靶基因。 调控总之，本提案中描述的实验将确定肌肉特性 类似的形状，方向和附件类型被指定，这将告知我们对脊椎动物的理解 人类肌肉发育的直系同源物，并导致治疗的发展。

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.