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Cytoskeletal Control of Microridge Morphogenesis on Mucosal Epithelial Cells of the Zebrafish Skin

斑马鱼皮肤粘膜上皮细胞微脊形态发生的细胞骨架控制

基本信息

批准号：
10393995
负责人：
Alvaro Sagasti
金额：
$ 1.07万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-08-01 至 2022-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10393995
关键词：
Actin-Binding Protein Actins Address Adopted Affect Animals Apical Architecture Bacterial Artificial Chromosomes Cell Shape Cell Surface Extensions Cell membrane Cell surface Cells Cellular Structures Complement Complex Cornea Cytokinesis Cytoskeletal Proteins Cytoskeleton Data Defect Development Disease Dry Eye Syndromes Elements Epithelial Epithelial Cells Esophagus Etiology F-Actin Fingerprint Fishes Genetic Recombination Glycoproteins Goals Grant Hydration status Image Individual Infection Intermediate Filaments Keratin Larva Lead Length Link Lubrication Membrane Microfilaments Modeling Molecular Molecular Conformation Morphogenesis Mucins Mucous Membrane Mucous body substance Myosin ATPase Myosin Type II Oral cavity Pattern Periderm Pharmacology Pharyngeal structure Play Process Property Protein Family Protein Isoforms Proteins Regulation Reporter Rho-associated kinase Role Shapes Signal Transduction Skin Structure Surface Testing Time Tissues United States National Institutes of Health Xerostomia Zebrafish base cellular microvillus conjunctiva drebrins experimental study eye dryness ezrin inhibitor/antagonist insight knock-down non-muscle myosin overexpression protein function rho GTP-Binding Proteins transcriptome sequencing

项目摘要

PROJECT SUMMARY To adopt forms optimized for their functions, individual cells sometimes project remarkably elaborate membrane protrusions, and even arrange them in complex patterns on their surfaces. To create and support membrane structures, the underlying cortical cytoskeleton must be arranged in specific conformations. The particular complement of cytoskeletal associated proteins, and the local regulation of their activity, thus determines the shape and pattern of cell membrane protrusions. To understand how cytoskeletal proteins together create unique cellular structures, we will study the formation of stunning actin-based structures on the surface of zebrafish skin cells called microridges. Microridges (or similar structures called microplicae) are found on most mucosal epithelial cells, which not only form the outer layer of fish skin, but also many of our own tissues, including the cornea, mouth, and parts of our gut. The microridge-covered surfaces of these cells display a glycoprotein calyx and adsorb mucins, suggesting that the unique structure of microridges is optimized for mucus retention. Mucus protects vulnerable epithelial tissues from abrasion and drying out, so understanding how microridges form could provide insight into the etiology of diseases affecting mucosal tissues, such as dry eye and dry mouth conditions. This proposal builds on successful descriptive and discovery-based studies supported by an NIH R21 grant that led to the identification of several new proteins in microridges. The experiments proposed here investigate mechanisms by which these specific proteins contribute to microridge morphogenesis, and, from a broader perspective, how they function as an ensemble to create the unique shapes and properties of microridges. In Aim 1 we will test the hypothesis that two proteins, Ezrin and Drebrin-like, initiate the first step of microridge morphogenesis, the formation of microvilli-like microridge precursors. Aim 2 investigates the interactions between F-actin and intermediate filaments (IFs) in microridges by testing if F-actin patterning determines IF patterning, and by testing the hypothesis that two candidate proteins, Envoplakin and Periplakin, link these cytoskeletal elements together. Finally, in Aim 3, live imaging, pharmacology, and molecular approaches will be used to characterize how myosin-based contraction and Rho GTPase signaling contribute to microridge morphogenesis. Collectively these studies will provide mechanistic insights into microridge morphogenesis, illuminate how cytoskeletal proteins together create elaborate cellular structures, and potentially point to how defects in epithelial morphogenesis contribute to diseases afflicting mucosal epithelia.

项目摘要为了采用最适合其功能的形式，单个细胞有时会投射出非常复杂的膜突起，甚至在它们的表面上以复杂的图案排列它们。建立和支持膜结构，下面的皮质细胞骨架必须安排在特定的构象。的细胞骨架相关蛋白的特定补体，以及它们活性的局部调节，决定了细胞膜突起的形状和模式。为了了解细胞骨架蛋白如何共同创造独特的细胞结构，我们将研究斑马鱼皮肤细胞表面惊人的肌动蛋白基结构称为微脊。微脊（或在大多数粘膜上皮细胞上发现了类似的结构，称为微褶，它们不仅形成了外膜，除了鱼的皮肤外，还有我们自己的许多组织，包括角膜、口腔和部分肠道。的这些细胞的微脊覆盖的表面显示出糖蛋白花萼并吸附粘蛋白，这表明微脊的独特结构被优化用于粘液保留。粘液保护脆弱的上皮组织从磨损和干燥，所以了解如何微脊形成可以提供深入了解的病因，影响粘膜组织的疾病，如干眼症和口干症。该提案建立在成功的描述性和基于发现的研究基础上，由NIH R21资助。从而在微脊中发现了几种新的蛋白质。这里提出的实验研究这些特定蛋白质有助于微脊形态发生的机制，并且，从更广泛的角度来看，透视图，它们如何作为一个整体来创造微脊的独特形状和特性。在目的1我们将验证两种蛋白质Ezrin和Drebrin-like启动微脊的第一步的假设形态发生，微绒毛样微脊前体的形成。目的2研究相互作用通过测试F-肌动蛋白模式是否决定了微脊中的IF，模式，并通过测试假设，两个候选蛋白，Envoplakin和Periplakin，连接这些细胞骨架元素在一起。最后，在目标3中，活体成像、药理学和分子方法将可用于表征肌球蛋白为基础的收缩和Rho GT3信号传导如何促进微脊形态发生总的来说，这些研究将为微脊形态发生提供机理上的见解，阐明细胞骨架蛋白如何共同创造精细的细胞结构，并可能指出如何上皮形态发生的缺陷会导致困扰粘膜上皮的疾病。