ApicalMembrane Modeling in Tubulogenesis

管发生中的顶膜建模

基本信息

批准号：
7858246
负责人：
VERENA GOBEL
金额：
$ 27.76万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-07-11 至 2013-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7858246
关键词：
Actins Adhesions Affect Apical Architecture Biochemical Biological Assay C. elegans genome Caenorhabditis elegans Cell Shape Cells Chimeric Proteins Chromosomes, Artificial, Yeast Collection Complement Complex Cosmids Cyst Cytoskeleton DNA Sequence Rearrangement Data Databases Defect Development Disease Electrons Enhancers Epithelial Epithelium Evaluation Family Family member Fluorescence Gene Order Genes Genetic Genetic Epistasis Genetic Polymorphism Genetic Screening Goals Human Intestines Kidney Libraries Life Link Maps Membrane Membrane Microdomains Microscopic Modeling Molecular Molecular Genetics Morphogenesis Movement Mutation Nature Nematoda Organ Organism Orthologous Gene Pathway interactions Pattern Phenocopy Phenotype Process Proteins Reagent Resources Role Shapes Site Sorting - Cell Movement Structural Models Surface System Testing Time Tissues Transgenic Organisms Tube Tubular formation Two-Hybrid System Techniques Vesicle Yeasts apical membrane base cell motility cellular microvillus computerized data processing ezrin feeding genetic analysis genome-wide in vivo insight intercalation loss of function membrane model migration moesin mutant overexpression protein structure radixin protein scaffold tool transmission process

项目摘要

DESCRIPTION (provided by applicant): Apical membrane modeling shapes organ lumina, defines membrane microdomains and leads cell movement during epithelial morphogenesis. The effector molecules and structural dynamics underlying these processes are still largely unknown in any species. We propose to analyze the molecular basis of apical membrane modeling in Caenorhabditis elegans tubulogenesis. The tubular organs of this transparent roundworm are derived from single-layered epithelia, providing a unique opportunity to study tube formation throughout development in a living organism. At the same time, its well characterized genetics allows for efficient molecular genetic analysis. We have shown that erm-1, the ancestral ortholog of the putative membrane-cytoskeleton linking ezrin, radixin, moesin (ERM) family, is required to shape the apical surfaces of the major internal organs of C. elegans, and to form intestinal microvilli and excretory canal canaliculi. We will attempt to dissect the mechanisms by which erm-1 exerts this morphogenetic function. In Aim 1, we will conduct an erm-1 genetic suppressor screen, as a specific approach to identify erm-1-interacting molecules involved in this process. Numerous proteins have been identified as interacting with vertebrate ERMs in diverse structural and signaling processes. The in vivo relevance of these data has been difficult to assess in the complex vertebrate system, given the redundancy among ERMs and their presumed developmental role. In Aim 2 we will conduct a candidate RNAi screen with orthologs of these proposed ERM-interacting molecules, to directly examine their effect on C. elegans morphogenesis. This screen utilizes C. elegans genome-wide databases, resources, and mutant collections. Aim 3 will confirm and characterize the genes identified in both screens by genetic, biochemical, morphological and functional assays; and attempt to define their effect on apical membrane modeling. Ordering these genes by epistasis should generate a construction kit for apical membrane modeling in epithelial morphogenesis. Given the fundamental nature of morphogenesis, and the unusually high conservation of ERMs among species, we expect to gain insight into processes relevant to human organ morphogenesis, particularly to intestinal and renal tube morphogenesis, and the diseases affecting these processes.

描述（由申请人提供）：顶部膜建模塑造器官腔，定义膜微域并导致上皮形态发生过程中的细胞运动。这些过程的效应分子和结构动力学在任何物种中仍然在很大程度上尚不清楚。我们建议分析秀丽隐杆线虫小管发生中顶膜建模的分子基础。这种透明的round虫的管状器官源自单层上皮，为在活生物体中的整个发育过程中提供了一个独特的机会来研究管的形成。同时，其良好的遗传学允许有效的分子遗传分析。我们已经表明，需要ERM-1，即塑造ezrin，radixin，moesin（Erm）家族的假定膜细胞骨架的祖先直系同源物，以塑造秀丽隐杆线虫主要内脏的顶端表面，并形成肠内微伏氏和排泄物的管道。我们将尝试剖析ERM-1发挥这种形态发生功能的机制。在AIM 1中，我们将进行ERM-1遗传抑制筛选，作为识别此过程中涉及的ERM-1相互作用分子的特定方法。在各种结构和信号过程中，许多蛋白质已被确定为与脊椎动物ERMS相互作用。考虑到ERMS之间的冗余及其推测的发育作用，这些数据的体内相关性很难在复杂的脊椎动物系统中评估。在AIM 2中，我们将进行候选RNAi屏幕，并具有这些提出的ERMEMTACTACTING分子的直系同源物，以直接检查其对秀丽隐杆线虫形态发生的影响。该屏幕利用秀丽隐杆线虫全基因组的数据库，资源和突变集收集。 AIM 3将通过遗传，生化，形态学和功能测定法确认并表征两个筛选中鉴定的基因。并尝试定义其对顶膜建模的影响。通过上学订购这些基因，应生成一个用于上皮形态发生中的顶膜建模的构造试剂盒。鉴于形态发生的基本性质以及物种之间异常的ERM保护，我们希望深入了解与人体器官形态发生相关的过程，尤其是与肠道和肾管形态发生以及影响这些过程的疾病。