MAST CELL GRANULES INDUCE ENDOTHELIAL CELL MOTILITY

肥大细胞颗粒诱导内皮细胞运动

• 批准号: 2029385
• 负责人: David Lagunoff
• 金额: $ 28.08万
• 依托单位: SAINT LOUIS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-01-01 至 1999-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2029385
• 关键词: cadherins; cell motility; confocal scanning microscopy; endothelin; enzyme inhibitors; fibroblast growth factor; gap junctions; granule; mast cell; phosphorylation; protein purification; protein structure function; protein tyrosine kinase; protein tyrosine phosphatase

DESCRIPTION (Adapted from Investigator's Abstract): This is a revised application examining the role of mast cells and adherent junctions in endothelial motility. Endothelial motility is critical to the formation of blood vessels during development, tumor formation, and in response to ischemia as well as an integral part of the response of the arterial wall to injury and atherosclerotic stimuli. The formation of blood vessels during regeneration, healing and tumor formation requires new vessels to form from old, and interruption of the continuity of existing endothelium requires re-establishment of the integrity of the endothelium. Both these processes require the release of endothelial cells from the joint inhibitions on cell movement and cell division characteristic of a stable, confluent sheet of endothelial cells. This investigator has observed that secretory granules from mast cells, when added to confluent monolayers of cultured endothelial cells, have the ability to restore motility to immotile endothelial cells in a confluent monolayer. Their evidence indicates that the component of the granule principally responsible for this activity is heparin proteoglycan. The free glycosaminoglycan form of heparin has no activity but can block the activity of granules. The regained motility of the cells induced by granules is associated with the formation of substantial gaps in the monolayer as the cells move. This proposal is based upon the finding that secretory granules from mast cells restore motility to immotile endothelial cells in confluent monolayers. The proposal hypothesizes that changes in proteins comprising adherent junctions (e.g., cadherin-5, catenins, p120, ZO-1, moesin, and ezrin) are responsible for the down regulation of motility following the establishment of confluent endothelial monolayers and the upregulation induced by mast cell granules or by bFGF. The extensively revised proposal uses a combination of confocal microscopy and protein quantification to examine the regulation of these proteins and to correlate them with changes in cell motility. Seven specific aims are proposed, including four new Aims. Aim 1 will correlate the display of cadherin-5 and associated proteins in adherent junctions with motility induced by mast cell granules and by bFGF. Aim 2 will examine the distribution of cadherin-5 in the cytoskeletal-bound state and the soluble cytoplasmic state. Four new aims will determine if changes observed in Aims 1 and 2 are related to tyrosine phosphorylation, develop tests of the functional state of adherent junctions to determine if motility is associated with weakening of the junctions, and determine in bFGF contributes to the effects of mass cell granules. A final Aim (7), will compare large vessel and microvascular endothelial cells.

描述(改编自调查人员摘要)：这是一个修订版 应用研究肥大细胞和贴壁连接在人类免疫缺陷中的作用 血管内皮细胞运动。血管内皮细胞的运动对血管的形成至关重要 血管在发育、肿瘤形成和对 缺血以及动脉壁对 损伤和动脉粥样硬化的刺激。血管的形成过程中 再生、愈合和肿瘤形成需要形成新的血管 陈旧，并中断现有内皮细胞的连续性需要 重建内皮细胞的完整性。这两个过程 要求内皮细胞从关节释放对细胞的抑制 一种稳定的、融合的薄片的运动和细胞分裂特性 内皮细胞。这位研究人员观察到，分泌颗粒 将肥大细胞加入培养的内皮细胞融合单层 细胞，有能力恢复运动到静止的内皮细胞在 融合的单层。他们的证据表明， 主要负责这一活动的颗粒是肝素蛋白多糖。 肝素的游离型糖胺多糖没有活性，但可以阻断 颗粒的活性。细胞运动功能的恢复。 颗粒与形成大量的空隙有关 随着细胞的移动而形成单层。这项建议是基于以下发现： 肥大细胞分泌颗粒恢复静止内皮细胞的运动 融合单层中的细胞。该提案假设，在 包含粘着连接的蛋白质(例如，钙粘附素-5，连环蛋白，p120， ZO-1、Moesin和Ezrin)负责运动性的下调 在建立融合内皮细胞单层和 肥大细胞颗粒或碱性成纤维细胞生长因子诱导的表达上调。广为流传 修订后的提案使用了共聚焦显微镜和蛋白质的组合 定量检测这些蛋白质的调节并进行相关分析 细胞运动性的改变。提出了七个具体目标， 包括四个新目标。目标1将钙粘附素-5和钙粘蛋白5的显示关联起来 肥大细胞诱导的黏附连接运动性相关蛋白 颗粒和碱性成纤维细胞生长因子。目标2将研究钙粘蛋白-5在脑内的分布 细胞骨架结合状态和可溶性细胞质状态。四个新的 AIMS将确定在AIMS 1和2中观察到的变化是否与 酪氨酸磷酸化，开发粘附物功能状态的测试 以确定运动性是否与脑电活动减弱有关 连接，并测定在碱性成纤维细胞生长因子对质量细胞的作用 颗粒。最终目标(7)，将比较大血管和微血管 内皮细胞。