CELL/CELL APPOSITION AND INFLAMMATORY EDEMA

细胞/细胞定位和炎症性水肿

• 批准号: 6390438
• 负责人: Douglas MICHAEL SHASBY
• 金额: $ 33.67万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-07-01 至 2003-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6390438
• 关键词: MDCK cell; cadherins; cell cell interaction; chromatography; confocal scanning microscopy; edema; electrophysiology; extracellular matrix; high endothelial venule; histamine; immunoprecipitation; inflammation; intercellular connection; phosphatase inhibitor; phosphorylation; polymerase chain reaction; protein tyrosine phosphatase; tissue /cell culture; transfection; vascular endothelium; MDCK cell; cadherins; cell cell interaction; chromatography; confocal scanning microscopy; edema; electrophysiology; extracellular matrix; high endothelial venule; histamine; immunoprecipitation; inflammation; intercellular connection; phosphatase inhibitor; phosphorylation; polymerase chain reaction; protein tyrosine phosphatase; tissue /cell culture; transfection; vascular endothelium

We have a long standing interest in understanding how inflammatory edema alters vascular and epithelial barriers. These barriers are dependent on the focal apposition of the cells to each other and to the underlying matrix. Our recently published work indicates that some agonists initially decrease endothelial and epithelial barrier function without increasing tension within the cells. More recent work indicates that histamine initially decreases focal apposition of HUVE cells at sites of cell-cell ahesion (measured as the impedance of a layer of cells on an electrode). In other recent work we found that the tyrosine phosphatase inhibitor, bPVphen, increases focal apposition of MDCK cells at cell-cell sites. This is associated with tyrosine phosphyorylation of E-cadherin. Cadherins are transmembrane proteins which contribute importanty to cell-cell apposition through homotypic binding at adherens junctions. ECV304 cells are a line of endothelial cells that do not express the endothelial cadherin, cadherin-5. When these cells are exposed to histamine they do not decrease focal apposition. When they are exposed to the tyrosine phosphatase inhibitor, bPVphen, they show a slow fall in impedance. In contrast, when the same cells are transfected with E-cadherin or cadherin-5 they show a prompt fall in cell-cell apposition with histamine. When the E-cadherin transfected cells are exposed to bPVphen, cell-cell apposition increases. When the cells transfected with cadherin-5 are exposed to bPVphen, cell-matrix, but not, cell-cell apposition increases, similar to what happens in HUVE cells. These observations implicate important effects of signalling on the cadherin and its uniquely associated proteins. We will use the transfected cells to examine effects of mutations in the cadherins on regulation of adherens junction function, which we can measure in real time.

我们对了解炎症性水肿如何改变血管和上皮屏障有很长的兴趣。这些障碍取决于细胞彼此的焦点和基础基质的焦点。 我们最近发表的工作表明，一些激动剂最初会降低内皮和上皮屏障功能，而不会增加细胞内部的张力。 最新的工作表明，组胺最初会降低HUVE细胞在细胞 - 细胞AHESIN的位点的焦点（以电极在电极上的阻抗为阻抗）。 在最近的其他工作中，我们发现酪氨酸磷酸酶抑制剂BPVPHEN增加了MDCK细胞在细胞细胞部位的局灶性分配。 这与电子钙黏着蛋白的酪氨酸磷酸化有关。 钙粘蛋白是跨膜蛋白，通过在粘附连接处的同型结合对细胞细胞的依赖有助于重要性。 ECV304细胞是一条未表达内皮钙粘蛋白Cadherin-5的内皮细胞系。 当这些细胞暴露于组胺时，它们不会降低局灶性。 当它们暴露于酪氨酸磷酸酶抑制剂BPVPHEN时，它们的阻抗下降缓慢。 相反，当用E-钙粘蛋白或钙粘蛋白5转染相同的细胞时，它们显示出与组胺的细胞细胞伴侣的迅速落下。 当E-钙粘着蛋白转染的细胞暴露于BPVPhen时，细胞 - 细胞的置换会增加。 当用钙粘蛋白5转染的细胞暴露于bpvphen时，细胞 - 矩阵，但不暴露于细胞 - 细胞的植入术增加，类似于HUVE细胞中发生的情况。这些观察结果暗示了信号传导对钙粘蛋白及其独特相关的蛋白质的重要​​影响。 我们将使用转染的细胞检查钙粘蛋白中突变对粘附连接函数调节的影响，我们可以实时测量。