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; 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（根据调查员的摘要改编）：这是一个修订版 申请检查肥大细胞和粘附连接在 内皮运动。 内皮运动对形成至关重要 发育过程中的血管，肿瘤形成以及对 缺血以及动脉壁反应的组成部分 损伤和动脉粥样硬化刺激。 在 再生，愈合和肿瘤形成需要新血管从 旧的和现有内皮连续性的中断需要 重新建立内皮的完整性。 这两个过程 需要从关节抑制细胞上释放内皮细胞 运动和细胞分裂特征的特征 内皮细胞。 该研究者观察到分泌颗粒 从肥大细胞中添加到培养的内皮的汇合单层中时 细胞，有能力恢复对及以疫苗内皮细胞的运动能力 汇合单层。 他们的证据表明 颗粒主要负责这项活动是肝素蛋白聚糖。 肝素的免费糖胺聚糖形式没有活动，但可以阻止 颗粒的活性。 通过 颗粒与形成大量差距有关 单层随着细胞的移动。 该提议基于以下发现 分泌颗粒从肥大细胞恢复运动性到Immotile内皮 汇合单层中的细胞。 该提议假设变化 包括粘附连接的蛋白质（例如Cadherin-5，Catenins，P120， ZO-1，Moesin和Ezrin）负责下调的调节 建立汇合的内皮单层和 肥大细胞颗粒或BFGF诱导的上调。 广泛 修订的建议结合了共聚焦显微镜和蛋白质 量化以检查这些蛋白质的调节并相关 它们随细胞运动的变化而变化。 提出了七个特定目标， 包括四个新目标。 AIM 1将使Cadherin-5的显示和 由肥大细胞引起的运动性粘附连接处的相关蛋白质 颗粒和BFGF。 AIM 2将检查Cadherin-5的分布 细胞骨架结合状态和可溶性细胞质状态。 四个新 目标将确定目标1和2中观察到的变化是否与 酪氨酸磷酸化，开发依从性功能状态的测试 确定运动性是否与弱化有关 连接并在BFGF中确定有助于质量细胞的影响 颗粒。 最终目标（7）将比较大容器和微血管 内皮细胞。