Planar Cell Polarity and the Cytoskeleton

平面细胞极性和细胞骨架

• 批准号: 7934690
• 负责人: ANDREAS JENNY
• 金额: $ 32.87万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2014-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7934690
• 关键词: Actins; Affect; Apical; Bardet-Biedl Syndrome; Biochemical Genetics; Biological Assay; Biological Models; Birds; Cell Communication; Cell Polarity; Cells; Cilia; Communication; Congenital Abnormality; Cytoskeleton; DNA Sequence Rearrangement; Data; Defect; Development; Dimerization; Dominant-Negative Mutation; Drosophila genome; Drosophila genus; Electrophoretic Mobility Shift Assay; Epithelial; Epithelium; Eye; Family; Feathers; Fishes; Generations; Genes; Genetic; Hair; Human; In Vitro; Knowledge; Labyrinth; Lead; Leukocytes; Ligands; Link; Literature; Lung; Mammals; Mediating; Mesenchymal; Microfilaments; Movement; Mucous body substance; Mutation; Nephronophthisis; Neural Tube Closure; Neural Tube Defects; Neural tube; Nuclear; Oncogenes; Organ; Organism; Pathway interactions; Phosphorylation; Photoreceptors; Physiological; Positioning Attribute; Process; Protein Family; Proteins; RNA Interference; Regulation; Research; Rho-associated kinase; Role; Rotation; Signal Transduction; Spinal Dysraphism; Stereocilium; Structure; System; Testing; Vertebrates; Wing; Work; Xenopus; Zebrafish; ascidian; base; cell determination; cell motility; fibrosarcoma; fly; gastrulation; genetic analysis; genome wide association study; genome-wide; in vivo; intercalation; knock-down; member; migration; mutant; novel; public health relevance; receptor; response; rho; sound; transcription factor

DESCRIPTION (provided by applicant): Planar cell polarity and the cytoskeleton Planar cell polarity (PCP) signaling regulates the establishment of polarity within the plane of an epithelium. The results of signaling are as diverse as the determination of cell fates, the generation of asymmetric, but highly aligned structures (e.g. stereocilia in the human inner ear or hairs on a fly wing), or the directional migration of cells during convergent extension during vertebrate gastrulation. PCP is governed by the non-canonical Fz/Planar Cell Polarity pathway, in which a Wnt signals through a Frizzled receptor leading to nuclear responses, as well as to cytoskeletal changes mediated by Rho Kinase. PCP signaling was originally discovered and is best studied in Drosophila, mainly because of the versatility of the fly as model system due to its low genetic redundancy. In Drosophila, PCP is essential for the orientation of the actin wing hairs and the polarization of the ommatidia in the eye, requiring highly coordinated movement of groups of photoreceptor cells (ommatidial rotation). Thus, key to PCP signaling in flies, and to convergent extension in vertebrates, are cytoskeletal rearrangements and cell migration processes. Central to these processes is Rho Kinase (Rock, Drok in Drosophila), mutations in which or dominant negative forms of which lead to ommatidial rotation/migration and convergent extension/neural tube defects in flies and fish, respectively. This proposal focuses on biochemical and genetic approaches to identify and characterize new PCP components and regulators of the cytoskeleton. In a genome wide screen we identified direct Drok substrates using a phosphorylation induced gel-shift assay. We will characterize the candidates using in vivo RNAi, mutational analysis and genetic interaction assays with known pathway components. We propose to study their mechanism of action with respect to Rho Kinase and PCP signaling in detail. Using the above strategies to work out mechanistic aspects of PCP signaling as well as to discover new links to cellular responses will extend our knowledge of early development. Due to the conservation of the PCP gene network in organisms as diverse as flies, ascidians and mammals, the proposed research will be of immediate importance for the understanding of the establishment of PCP and related morphogenetic processes in vertebrates. PUBLIC HEALTH RELEVANCE: The mechanism of communication between cells of the fruitfly Drosophila and humans is well conserved and very similar. We make use of Drosophila to study how cell communication affects cell movement. Lack of such communication can lead to severe birth defects such as open neural tubes (e.g. spina bifida).

描述（由申请人提供）：平面细胞极性和细胞骨架平面细胞极性（PCP）信号传导调节上皮平面内的极性建立。信号传导的结果与细胞命运的确定，不对称但高度排列的结构的产生（例如，人体内耳或蝇翼上的头发的立体胶质）或在脊椎动物胃肠道过程中收敛延伸过程中的定向迁移。 PCP受非典型的FZ/Planar细胞极性途径的控制，其中WNT通过卷曲的受体发出信号，导致核反应以及Rho激酶介导的细胞骨架变化。 PCP信号最初是在果蝇中发现的，并且最好在果蝇中进行研究，这主要是由于果蝇作为模型系统的多功能性，这是由于其遗传冗余低。在果蝇中，PCP对于肌动蛋白机翼头发的方向和眼睛中的Ommatidia的极化至关重要，需要高度协调的光感受器细胞（胶状旋转）运动。因此，果蝇中的PCP信号传导以及脊椎动物的收敛延伸的关键是细胞骨架重排和细胞迁移过程。这些过程的核心是Rho激酶（岩石，果蝇中的Drok），其中分别导致果蝇和鱼类中产生跨性旋转/迁移和收敛延伸/神经管缺陷的突变。该建议着重于生化和遗传方法，以识别和表征细胞骨架的新PCP组件和调节剂。在基因组宽屏幕中，我们使用磷酸化诱导的凝胶转移测定法确定了直接Drok底物。我们将使用体内RNAi，突变分析和与已知途径成分的遗传相互作用分析来表征候选者。我们建议研究他们在Rho激酶和PCP信号方面的作用机理。使用上述策略来确定PCP信号的机理方面，以及发现与细胞反应的新链接，将扩展我们对早期发展的了解。由于在像苍蝇一样多样化的生物体中保存了PCP基因网络，因此拟议的研究对于理解脊椎动物中PCP的建立以及相关的形态学过程至关重要。公共卫生相关性：果蝇果蝇和人类细胞之间交流的机制是充分保守的，而且非常相似。我们利用果蝇研究细胞通信如何影响细胞运动。缺乏这种交流会导致严重的先天缺陷，例如开放神经管（例如脊柱裂）。