New vascular guidance signaling pathway

新的血管引导信号通路

基本信息

批准号：
7248788
负责人：
DEAN Yaw LI
金额：
$ 31.89万
依托单位：
UNIVERSITY OF UTAH
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-07-01 至 2009-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7248788
关键词：
Ablation Actins Adhesions Affect Alkaline Phosphatase Anatomy Apoptosis Applications Grants Architecture Axon Binding Biochemical Biological Assay Biological Neural Networks Biology Blood Vessels Cell Adhesion Cells Cellular Assay Cellular biology Complex Cues Data Development Distal Drosophila sli protein Endothelial Cells Family Family member Fibroblast Growth Factor Fibroblast Growth Factor 2 Focal Adhesions Gene Expression Genes Genetic Programming Growth Cones Heart Immunoprecipitation Integrins Kidney Knockout Mice Lead Ligands Link Lung Mammalian Cell Mediating Mediator of activation protein Modeling Molecular Morphology Movement Mus Nerve Nervous system structure Neural tube Organ Pathway interactions Pattern Protein Binding Protein Family Proteins Retina Role Signal Pathway Signal Transduction Stress Fibers Structure Tube Vascular Endothelial Growth Factors Vascular Endothelium Vascular System Yeasts angiogenesis axonal guidance cell motility domain mapping in vivo member migration novel paxillin polymerization receptor relating to nervous system research study yeast two hybrid system

项目摘要

DESCRIPTION (provided by applicant): The regimented and conserved pattern of the vascular network suggests that specific genetic programs coordinate its formation. Both the nervous and vascular systems begin as central axial structures that send sprouts to distal targets and form complex, highly reproducible and often superimposable networks. The development of the neural network is regulated by combinations of attractive and repulsive cues that guide growth cones. Secreted Slit proteins provide repulsive signals through their cognate Robo receptors and are critical for regulating whether axons cross the midline. Recently, we found that the vascular endothelium exclusively expresses a novel member of the Robo receptor family, Robo4. Slit-Robo4 signaling blocks the migration of cells to fibroblast growth factors and vascular endothelial growth factor. Here, we propose to understand the role of Robo signaling in vascular guidance. In Specific Aim 1, we will investigate the role of Robo4 signaling in endothelial sprouting. Using cell biology assays, we will determine whether the function of Slit-Robo4 signaling in endothelial cell biology is analogous to the role of Slit-Robo signaling in axonal guidance. We predict that Slit-Robo4 signaling will disrupt endothelial migration, sprouting, adhesion and cytoskeletal architecture but have no effect on proliferation or tube formation. Specific Aim 2 will examine the molecular mechanism of Robo4 signaling. Using yeast-two hybrid analyses and immunoprecipitations, we will identify intracellular proteins that bind to Robo4 and mediate its downstream signaling cascade. Our studies have revealed a potential link between Robo4 receptors and the paxillin family of proteins. We will define the biochemical interactions between Robo4 and downstream proteins, and the functional roles of these interactions. In Specific Aim 3, we will examine the role of Robo4 in vascular development using knockout mice. Preliminary data suggest that ablating Robo4 results in increased vascular sprouting around and within the neural tube. Together, the studies outlined in this grant application seek to demonstrate that nerves and vessels exploit a common signaling mechanism, the Slit-Robo pathway, for guiding the formation of their respective networks along specific tracts.

描述（由申请人提供）：血管网络的严格且保守的模式表明特定的遗传程序协调其形成。神经和血管系统都始于中轴结构，将芽发送到远端目标并形成复杂、高度可重复且通常可叠加的网络。神经网络的发育受到引导生长锥的吸引和排斥线索的组合的调节。分泌的 Slit 蛋白通过其同源 Robo 受体提供排斥信号，对于调节轴突是否穿过中线至关重要。最近，我们发现血管内皮专门表达 Robo 受体家族的一个新成员，Robo4。 Slit-Robo4 信号传导阻断细胞向成纤维细胞生长因子和血管内皮生长因子的迁移。在这里，我们建议了解 Robo 信号在血管引导中的作用。在具体目标 1 中，我们将研究 Robo4 信号传导在内皮细胞萌芽中的作用。通过细胞生物学分析，我们将确定 Slit-Robo4 信号在内皮细胞生物学中的功能是否类似于 Slit-Robo 信号在轴突引导中的作用。我们预测 Slit-Robo4 信号传导将破坏内皮迁移、出芽、粘附和细胞骨架结构，但对增殖或管形成没有影响。具体目标 2 将检查 Robo4 信号传导的分子机制。使用酵母-两种杂交分析和免疫沉淀，我们将鉴定与 Robo4 结合并介导其下游信号级联的细胞内蛋白质。我们的研究揭示了 Robo4 受体与桩蛋白家族之间的潜在联系。我们将定义 Robo4 和下游蛋白质之间的生化相互作用，以及这些相互作用的功能作用。在特定目标 3 中，我们将使用基因敲除小鼠研究 Robo4 在血管发育中的作用。初步数据表明，消融 Robo4 会导致神经管周围和内部的血管萌发增加。总之，本次拨款申请中概述的研究旨在证明神经和血管利用共同的信号传导机制（Slit-Robo 通路）来指导沿着特定束形成各自的网络。