Mechanosensitive signaling and cell adhesion during migration

迁移过程中的机械敏感信号传导和细胞粘附

基本信息

批准号：
7409323
负责人：
Gregory Weber
金额：
$ 4.68万
依托单位：
UNIVERSITY OF VIRGINIA
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-03-15 至 2010-03-14
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7409323
关键词：
Actins Address Adhesions Adhesives Adult Affect Arts Atomic Force Microscopy Behavior Biological Assay Biological Models Biomechanics C cadherin Cadherins Cell Adhesion Cell Polarity Cell-Cell Adhesion Cell-Matrix Junction Cells Chemotaxis Chimeric Proteins Complex Condition Confocal Microscopy Cues Cultured Cells Development Disease Elements Environment Exhibits Family Fibronectins Fluorescence Resonance Energy Transfer Focal Adhesions Green Fluorescent Proteins Guanosine Triphosphate Phosphohydrolases Immigration In Vitro Individual Integrins Investigation Life Ligands Link Mechanics Mediating Microscopy Modeling Molecular Morphogenesis Movement Pathology Physiology Platelet-Derived Growth Factor Process Regulation Resolution Role Signal Transduction Speed System Techniques Testing Time Tissues Traction Xenopus cell motility cell type cellular imaging computerized data processing extracellular in vivo insight magnetic beads migration response rho spatiotemporal

项目摘要

DESCRIPTION (provided by applicant): Cell migration is a complex process that involves both physical adhesive mechanics and intracellular signaling, which are profoundly interdependent on one another. Mechanisms of cell migration have been studied intensively using single cells cultured in vitro, but our understanding of cell migration in vivo remains limited. Cell migration in vivo often occurs in multicellular arrays that produce bulk tissue movements critical to morphogenesis, normal adult physiology, and various pathologies. While the function of mechanical signaling generated through cell-matrix traction forces has frequently been the subject of investigation, mechanosensitive responses to tension in cell-cell adhesions and the subsequent effects of these adhesive contacts on cell migration have not been elucidated. I will test the hypothesis that mechanical signaling, initiated by cadherin-mediated cell-cell adhesions in intact migratory tissues, has a central role in coordinating cell polarization and directional migration of individual cells to direct intact tissue movements. Mechanisms of cell migration are widely conserved between different species, tissues, and cell types. The migratory mesendoderm that traverses the fibronectin matrix of the blastocoel roof during Xenopus gastrulation will be used as a model system for these studies. This model provides an ideal system in which to study normal in vivo migratory processes because the motile behaviors of the intact tissue are retained when cultured in vitro and thus, the system is accessible to a variety of experimental manipulations. There are four specific aims proposed to address the significance of cell-cell adhesion in the propagation of mechanical signals that regulate mesendoderm cell polarity and migration. In specific aim 1, mesendoderm explants and a magnetic bead "pull" assay will be used to establish the role of biomechanical tension on C-cadherin dependent adhesions as an important regulator of polarization and directional migration in this tissue. Cadherins and integrins provide a physical link between extracellular ligands and internal cytoskeletal elements. GFP fusion proteins will be used to track cytoskeletal dynamics as a consequence of mechanical signaling initiated by cadherin adhesions in specific aim 2. Actin cytoskeletal dynamics involved in migration is signaled by spatiotemporal activation of the small Rho family GTPases. In specific aim 3, FRET analysis will be used to examine the local subcellular activation of Rho family GTPases as a result of C-cadherin and a5 (31 integrin-dependent mechanosensitive cell signaling. Finally, traction force microscopy and reflection contrast microscopy approaches will be used in specific aim 4 to determine the specific force relationship between cell-matrix adhesions and C-cadherin cell-cell contacts in migrating mesendoderm.

描述（由申请人提供）：细胞迁移是一个复杂的过程，涉及物理粘合力力学和细胞内信号传导，它们彼此之间非常相互依存。已经使用在体外培养的单细胞进行了深入研究细胞迁移的机制，但是我们对体内细胞迁移的理解仍然有限。体内细胞迁移通常发生在多细胞阵列中，产生对形态发生，正常成人生理学和各种病理至关重要的散装组织运动。虽然通过细胞基质牵引力产生的机械信号传导的功能经常是研究的主题，但机械敏感的反应对细胞 - 细胞粘连中的张力的反应以及这些粘合剂接触对细胞迁移的随后影响尚未阐明。我将检验以下假设：由钙粘蛋白介导的细胞 - 细胞粘附在完整迁移组织中引发的机械信号传导在协调细胞极化和单个细胞的方向迁移以导致完整的组织运动方面具有核心作用。细胞迁移的机制在不同的物种，组织和细胞类型之间得到了广泛保守。在异武肠胃肠道期间，跨胚层屋顶的纤连蛋白基质的迁移序列胚层将用作这些研究的模型系统。该模型提供了一个理想的系统，可以在其中研究正常的体内迁移过程，因为在体外培养时保留了完整组织的运动行为，因此，该系统可用于多种实验性操作。提出了四个特定的目的，以解决细胞 - 细胞粘附在调节中胚层细胞极性和迁移的机械信号传播中的重要性。在特定的目标1中，将使用中胚层外植体和磁珠“拉动”测定法来确定生物力学张力对C-钙粘着蛋白依赖性粘附的作用，作为该组织中极化和定向迁移的重要调节剂。钙粘蛋白和整联蛋白在细胞外配体和内部细胞骨架元素之间提供物理联系。 GFP融合蛋白将用于跟踪特定目标中钙粘蛋白粘附引发的机械信号传导的结果。在特定目标3中，将使用FRET分析来检查Rho家族GTPases的局部亚细胞激活，这是C-钙粘着蛋白和A5的结果（31个整合素依赖蛋白依赖性的机械敏感细胞信号传导。最后，牵引力显微镜和反射对比显微镜方法将是将是在特定目标4中用于确定迁移的胚胚层中细胞 - 矩阵粘连与C-钙粘蛋白细胞 - 细胞接触之间的特定力关系。