Celsr1-mediated planar cell polarity: defining the adhesive interface and mechanisms of asymmetry

Celsr1 介导的平面细胞极性：定义粘附界面和不对称机制

• 批准号: 9328279
• 负责人: Sara N Stahley
• 金额: $ 5.67万
• 依托单位: PRINCETON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2020-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9328279
• 关键词: Adhesions; Adhesives; Architecture; Biochemical; Biochemistry; Biological Assay; Biological Models; Cadherins; Cardiomyopathies; Cell Communication; Cell Culture Techniques; Cells; Complex; Cytoplasmic Protein; Defect; Development; Disease; Drosophila genus; Dsh protein; Embryo; Embryonic Development; Employee Strikes; Epidermis; G-Protein-Coupled Receptors; Genetic; Genetic Techniques; Goals; Hair; Health; Heart; Heart Abnormalities; Human; Imaging Techniques; In Vitro; Individual; Knowledge; Lateral; Lighting; Maintenance; Mammals; Mediating; Membrane; Microscopy; Molecular; Molecular Biology; Mus; Mutagenesis; Mutation; N-terminal; Neural Tube Defects; Neurons; Optics; Organism; Pathway interactions; Pattern; Phenotype; Protein Biochemistry; Proteins; Recruitment Activity; Resolution; Role; Signal Pathway; Signal Transduction; Skin; Spinal Dysraphism; Structure; System; Testing; Tissues; Variant; ciliopathy; developmental disease; extracellular; fly; genetic analysis; in vivo; insight; member; mutant; novel; planar cell polarity; polarized cell; reconstruction; targeted treatment; temporal measurement

Project Summary Planar cell polarity (PCP) is a core cell contact and signaling pathway that is largely conserved from Drosophila to mammals, and is fundamental to early development and tissue organization in complex, multicellular organisms. The long-term goal of this study is to further our understanding of how central PCP protein, Celsr1, mediates adhesive interactions required for the establishment and maintenance of PCP. Acting orthogonally to apico-basal polarity, PCP is the collective and organized polarization of cells along a tissue plane. A striking example of PCP is the ordered alignment of body hairs on mammalian skin. Core components of this pathway include: Celsr (Flamingo/Fmi), Frizzled (Fz), Vangl (VanGogh/Vang), Dishevelled (Dvl), Prickle (Pk) and Diego (Dgo). A hallmark feature of PCP is the asymmetric localization of Fzd/Dv/Dgo and Vangl/Pk at cell borders within a junctional complex organized via intercellular interactions of Celsr, a large atypical cadherin and member of the GPCR super-family. Disrupting Celsr function in mice leads to a range of detrimental phenotypes including neural tube defects, abnormal hair patterning and embryonic lethality. Further, multiple Celsr mutations in humans have been associated with neural tube defects and diseases such as spina bifida and cardiomyopathies. While important developmental roles for Celsr at the tissue-level have been established with mouse and fly studies, an understanding of Celsr function at the molecular and cell level is lacking. To elucidate how PCP asymmetry is achieved, we need to define the mechanisms of Celsr1 extracellular adhesion. The mouse skin is an ideal model system to investigate the conserved mechanisms of PCP and our lab has recently established a spontaneously polarizing in vitro, organotypic model system for assaying PCP asymmetry and function. Super-resolution imaging techniques such as structured illumination microscopy (SIM) and direct stochastic optical reconstruction microscopy (dSTORM) further add to our robust toolbox to study epidermal PCP. The overall goal of this proposal is to investigate how the principle PCP protein, Celsr1, mediates extracellular adhesion to coordinate functional PCP. We hypothesize that Celsr1 cadherin repeats mediate intercellular and lateral adhesive interactions required for asymmetrical PCP protein localization and function. Understanding how individual cell-cell interactions at the molecular level contribute to tissue structure and function will fill a substantial and fundamental knowledge gap in the PCP field. This proposal integrates molecular biology, biochemistry and advanced microscopy approaches to elucidate mechanisms of PCP asymmetry and function in vitro and in vivo. Completion of these studies will provide novel insight into the mechanisms the regulate PCP in the mammalian epidermis at the most fundamental level and illuminate our conceptual understanding of PCP complex formation and function.

项目概要 平面细胞极性（PCP）是核心细胞接触和信号传导途径，在很大程度上是保守的 果蝇对哺乳动物来说，是复杂的早期发育和组织组织的基础， 多细胞生物。这项研究的长期目标是进一步了解中央 PCP 如何 蛋白质 Celsr1 介导建立和维持 PCP 所需的粘附相互作用。演戏 与顶端-基底极性正交，PCP 是沿组织的细胞集体和有组织的极化 飞机。 PCP 的一个显着例子是哺乳动物皮肤上体毛的有序排列。核心部件 该途径的包括：Celsr (Flamingo/Fmi)、Frizzled (Fz)、Vangl (VanGogh/Vang)、Disheveled (Dvl)、Prickle （Pk）和迭戈（Dgo）。 PCP 的一个标志性特征是 Fzd/Dv/Dgo 和 Vangl/Pk 的不对称定位 通过 Celsr（一种大型非典型细胞）的细胞间相互作用组织的连接复合体内的细胞边界 钙粘蛋白和 GPCR 超家族的成员。破坏小鼠的 Celsr 功能会导致一系列 有害的表型包括神经管缺陷、毛发图案异常和胚胎致死。 此外，人类的多种 Celsr 突变与神经管缺陷和疾病有关，例如 如脊柱裂和心肌病。虽然 Celsr 在组织水平上具有重要的发展作用 通过小鼠和果蝇研究建立了对 Celsr 功能在分子和细胞水平上的了解 缺乏。为了阐明 PCP 不对称性是如何实现的，我们需要定义 Celsr1 的机制 细胞外粘附。小鼠皮肤是研究保守机制的理想模型系统 PCP 和我们的实验室最近建立了一个自发极化的体外器官模型系统 测定 PCP 不对称性和功能。超分辨率成像技术，例如结构照明 显微镜 (SIM) 和直接随机光学重建显微镜 (dSTORM) 进一步增强了我们的稳健性 研究表皮 PCP 的工具箱。该提案的总体目标是研究原则 PCP 如何 蛋白质 Celsr1 介导细胞外粘附以协调功能性 PCP。我们假设 Celsr1 钙粘蛋白重复序列​​介导不对称 PCP 蛋白所需的细胞间和横向粘附相互作用 定位和功能。了解分子水平上单个细胞与细胞之间的相互作用如何有助于 组织结构和功能将填补 PCP 领域的重大基础知识空白。这 该提案整合了分子生物学、生物化学和先进的显微镜方法来阐明 PCP 不对称性及其体外和体内功能的机制。完成这些研究将提供新颖的 深入了解哺乳动物表皮中 PCP 的调节机制，以及 阐明我们对 PCP 复合物形成和功能的概念理解。