CONTROL OF MUSCLE PROTEIN SYNTHESIS DURING MYOGENESIS

肌生成过程中肌肉蛋白合成的控制

基本信息

批准号：
8051021
负责人：
CHARLES P. EMERSON
金额：
$ 1.18万
依托单位：
BOSTON BIOMEDICAL RESEARCH INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-05-01 至 2011-09-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8051021
关键词：
Biological Assay Birds Cell Lineage Cell surface Cells Cloning Complementary DNA Computer Analysis Cultured Cells Development Disease Electroporation Embryo Enhancers Erinaceidae Extracellular Matrix Family Fibroblast Growth Factor Gene Expression Gene Targeting Genes Genetic Genetic Transcription Homologous Gene Investigation Knowledge Lead Maintenance Maps Mediating Molecular Molecular Genetics Mus Muscle Muscle Proteins Myoblasts Neural Crest Nuclear Organ Process Protein Biosynthesis Proteoglycan Regulation Regulator Genes Regulatory Element Reporter Genes Role Sclerotome Signal Pathway Signal Transduction Signal Transduction Pathway Signaling Molecule Site-Directed Mutagenesis Skeletal Muscle Somites Staging Stem cells Sulfatases Surface Ectoderm Tertiary Protein Structure Testing Tissues Transcriptional Activation Transfection Transgenic Organisms Trauma base embryo culture gene function genetic analysis genetic regulatory protein human tissue keratan sulfate proteoglycan myogenesis new technology notochord novel progenitor relating to nervous system repaired research study skeletal somitogenesis stem cell division transcription factor

项目摘要

Skeletal muscle provides a unique opportunity to investigate molecular genetic mechanisms that control specification of stem cell lineages within the cellular complexity of the vertebrate embryo. The proposed studies investigate developmental mechanisms that regulate the myogenic regulatory genes, MyfS and MyoD, which control the specification of skeletal muscle lineages from somite progenitors. Experiments focus specifically on the molecular mechanisms by which Sonic hedgehog (Shh) and Wnt signal molecules, produced by surrounding tissues, induce the transcriptional activation MyfS and MyoD in somitic progenitor cells, and thus initiate the process of myogenic cell specification. Complementary approaches have been developed to investigate these mechanisms in mouse and avian embryos. First, MyoD and MyfS transcriptional enhancers will be characterized using transgenic and transfection reporter gene assays, in combination with site-directed mutagenesis and nuclear factor footprinting, to map regulatory sequences and identify interacting transcription factors hypothesized to mediate Shh and Wnt induction of MyoD and MyfS.These studies undertake to distinguish whether the Gli and (3-catenin/LEFtranscription factors, which mediate Shh and Wnt signal transduction, interact directly with MyoD and MyfS enhancers to control their activation, or alternatively,whether these signals function upstream to regulate somite-specific transcription factors. Second, the developmental functions of a set of somite-activated regulatory genes, identified in differential cDNA cloning screens, will be investigated by antisense inhibition and misexpression analyses in avian embryos and by gene targeting and genetic analyses in the mouse embryo to determine their specific functions in MyoD and MyfS regulation during somite formation. These studies will focus on a family of two novel Sulfatases that were identified in our screen for somite-activated genes and are required for activation of MyoD in somites of avian embryos. These Sulfatases are hypothesized to desulfate proteoglycans in the extracellular matrix, thus controlling the localized activities of developmental signaling molecules essential for MyoD activation. Other novel somite-activated regulatory genes, identified in these screens, will be investigated to define their functions in the control of somite formation, Shh signal transduction, and MyoD and MyfS activation. The findings of these investigations are expected to lead to the discovery of novel developmental regulatory genes that control the specification of myogenic cells and other lineages in vertebrate embryos. Such knowledge will define fundamentalmechanisms that control of stem cell formation, and this information will likely lead to the development of new technologies to promote stem cell renewal in the repair of human tissues and organs damaged by disease and trauma.

骨骼肌提供了一个独特的机会来研究控制干细胞规范的分子遗传机制在脊椎动物胚胎的细胞复杂性内谱系。拟议的研究研究了发展机制调节肌源性调节基因，MyFS和Myod，这些基因控制了体系的骨骼肌谱系的规范祖先。实验专门针对声音刺猬（SHH）和Wnt信号分子的分子机制，由周围的组织产生，诱导躯体祖细胞中的转录激活myf和myod，从而启动肌原细胞规范的过程。已经开发了互补的方法来研究这些机制鼠标和禽类胚胎。首先，将使用转基因和转染来表征Myod和MyF的转录增强器报告基因测定与位置定向的诱变和核因子足迹结合使用，以绘制调节序列和确定相互作用的转录因子，假设介导Myod和MyF的SHH和Wnt诱导。为了区分GLI和（介导SHH和Wnt信号转导的3-catenin/leftranscrions因子，相互作用）直接使用Myod和MyFS增强器来控制其激活，或者，这些信号是否在上游运行到调节特异性转录因子。其次，一组体积激活的调节基因的发育功能，在差异cDNA克隆筛网中鉴定，将通过反义抑制和Misexpression分析进行研究胚胎以及通过基因靶向和遗传分析在小鼠胚胎中确定其在MYOD和MYF中的特定功能在体积形成期间的调节。这些研究将集中于我们屏幕上发现的两个新型硫酸酶的家族在鸟类胚胎体中激活MYOD所必需的体激活基因。这些硫酸酶被认为在细胞外基质中脱硫蛋白聚糖，从而控制发育信号分子的局部活性 Myod激活至关重要。在这些筛选中鉴定出的其他新型的体积激活的调节基因将研究到定义它们在控制体形成，SHH信号转导以及MYOD和MYF激活中的功能。结果预计这些研究将导致发现新的发育调节基因，以控制的规范脊椎动物胚胎中的肌原细胞和其他谱系。这样的知识将定义控制茎的基本机制细胞形成，这些信息可能会导致开发新技术以促进干细胞更新修复疾病和创伤受损的人体组织和器官。