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

• 批准号: 9982348
• 负责人: Alvaro Sagasti
• 金额: $ 29.26万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 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/9982348
• 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; Elements; Epithelial; Epithelial Cells; Epithelium; Esophagus; Etiology; Expression Profiling; Eye diseases; 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

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 为了采用针对其功能优化的表格，各个单元有时会非常精心阐述 膜突起，甚至在其表面上以复杂的图案排列。创建和支持 膜结构，潜在的皮质细胞骨架必须以特定的构型排列。这 细胞骨架相关蛋白的特殊补体及其活性的局部调节，因此 确定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建独特的细胞结构，我们将研究形成 斑马鱼皮细胞表面上的令人惊叹的基于肌动蛋白的结构称为微型树木。微矿石（或 在大多数粘膜上皮细胞上都发现了称为微斑的类似结构，这不仅形成了外部 鱼皮层，还有我们自己的许多组织，包括角膜，嘴和肠道的部分。这 这些细胞的Microidge覆盖表面显示糖蛋白花萼和吸附粘蛋白，这表明 优化微型树木的独特结构以保留粘液。粘液保护脆弱的上皮组织 从磨损和干燥中，因此了解微型车队的形成如何提供对病因的见解 影响粘膜组织的疾病，例如干眼症和口干条件。 该提案以NIH R21赠款支持的成功描述性和基于发现的研究为基础 这导致了微型树木中几种新蛋白质的鉴定。这里提出的实验调查 这些特定蛋白有助于微岛形态发生的机制，并从更广泛的 透视图，它们如何充当一个合奏，以创建微型泳衣的唯一形状和属性。在 AIM 1我们将测试两个蛋白质Ezrin和Drebrin样启动Microide的第一步的假设 形态发生，形成微绒毛样微岛前体。 AIM 2调查互动 通过测试F-肌动蛋白是否决定是否确定是否是否确定是否是否 通过测试两个候选蛋白Envoplakin和Periplakin的假设，将其联系起来 细胞骨架元素在一起。最后，在AIM 3中，实时成像，药理学和分子方法将 用于表征基于肌球蛋白的收缩和Rho GTPase信号如何有助于Microdide 形态发生。这些研究集体将提供有关微岛形态发生的机械见解， 阐明细胞骨架蛋白如何共同创建精美的细胞结构，并有可能指出 上皮形态发生的缺陷有助于困扰粘膜上皮的疾病。