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Journal of Cell Science, Vol 114, Issue 7 1343-1355, Copyright © 2001 by Company of Biologists
JOURNAL ARTICLES |
B Wojciak-Stothard, S Potempa, T Eichholtz and AJ Ridley
Ludwig Institute for Cancer Research, Royal Free and University College School of Medicine, London W1W 7BS, UK. anne@ludwig.ucl.ac.uk
Endothelial permeability induced by thrombin and histamine is accompanied by actin stress fibre assembly and intercellular gap formation. Here, we investigate the roles of the &Rgr; family GTPases Rho1, Rac1 and Cdc42 in regulating endothelial barrier function, and correlate this with their effects on F-actin organization and intercellular junctions. RhoA, Rac1 and Cdc42 proteins were expressed efficiently in human umbilical vein endothelial cells by adenovirus-mediated gene transfer. We show that inhibition of &Rgr; prevents both thrombin- and histamine-induced increases in endothelial permeability and decreases in transendothelial resistance. Dominant-negative RhoA and a &Rgr; kinase inhibitor, Y-27632, not only inhibit stress fibre assembly and contractility but also prevent thrombin- and histamine-induced disassembly of adherens and tight junctions in endothelial cells, providing an explanation for their effects on permeability. In contrast, dominant-negative Rac1 induces permeability in unstimulated cells and enhances thrombin-induced permeability, yet inhibits stress fibre assembly, indicating that increased stress fibre formation is not essential for endothelial permeability. Dominant-negative Cdc42 reduces thrombin-induced stress fibre formation and contractility but does not affect endothelial cell permeability or responses to histamine. These results demonstrate that &Rgr; and Rac act in different ways to alter endothelial barrier function, whereas Cdc42 does not affect barrier function.
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 D. GRATZINGER, S. CANOSA, B. ENGELHARDT, and J. A. MADRI Platelet endothelial cell adhesion molecule-1 modulates endothelial cell motility through the small G-protein Rho FASEB J, August 1, 2003; 17(11): 1458 - 1469. [Abstract] [Full Text] [PDF]
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M. Holinstat, D. Mehta, T. Kozasa, R. D. Minshall, and A. B. Malik Protein Kinase C{alpha}-Induced p115RhoGEF Phosphorylation Signals Endothelial Cytoskeletal Rearrangement J. Biol. Chem., August 1, 2003; 278(31): 28793 - 28798. [Abstract] [Full Text] [PDF]
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J. Qiao, F. Huang, and H. Lum PKA inhibits RhoA activation: a protection mechanism against endothelial barrier dysfunction Am J Physiol Lung Cell Mol Physiol, June 1, 2003; 284(6): L972 - L980. [Abstract] [Full Text] [PDF]
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B. Wojciak-Stothard and A. J. Ridley Shear stress-induced endothelial cell polarization is mediated by Rho and Rac but not Cdc42 or PI 3-kinases J. Cell Biol., April 28, 2003; 161(2): 429 - 439. [Abstract] [Full Text] [PDF]
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S. K. Quadri, M. Bhattacharjee, K. Parthasarathi, T. Tanita, and J. Bhattacharya Endothelial Barrier Strengthening by Activation of Focal Adhesion Kinase J. Biol. Chem., April 4, 2003; 278(15): 13342 - 13349. [Abstract] [Full Text] [PDF]
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N. M. Rubenstein, Y. Guan, P. L. Woo, and G. L. Firestone Glucocorticoid Down-regulation of RhoA Is Required for the Steroid-induced Organization of the Junctional Complex and Tight Junction Formation in Rat Mammary Epithelial Tumor Cells J. Biol. Chem., March 14, 2003; 278(12): 10353 - 10360. [Abstract] [Full Text] [PDF]
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G. Benais-Pont, A. Punn, C. Flores-Maldonado, J. Eckert, G. Raposo, T. P. Fleming, M. Cereijido, M. S. Balda, and K. Matter Identification of a tight junction-associated guanine nucleotide exchange factor that activates Rho and regulates paracellular permeability J. Cell Biol., March 3, 2003; 160(5): 729 - 740. [Abstract] [Full Text] [PDF]
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S. W. Klarenbach, A. Chipiuk, R. C. Nelson, M. D. Hollenberg, and A. G. Murray Differential Actions of PAR2 and PAR1 in Stimulating Human Endothelial Cell Exocytosis and Permeability: The Role of Rho-GTPases Circ. Res., February 21, 2003; 92(3): 272 - 278. [Abstract] [Full Text] [PDF]
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G. P. van Nieuw Amerongen, P. Koolwijk, A. Versteilen, and V. W.M. van Hinsbergh Involvement of RhoA/Rho Kinase Signaling in VEGF-Induced Endothelial Cell Migration and Angiogenesis In Vitro Arterioscler. Thromb. Vasc. Biol., February 1, 2003; 23(2): 211 - 217. [Abstract] [Full Text] [PDF]
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R. Ruiz-Velasco, C. C. Lanning, and C. L. Williams The Activation of Rac1 by M3 Muscarinic Acetylcholine Receptors Involves the Translocation of Rac1 and IQGAP1 to Cell Junctions and Changes in the Composition of Protein Complexes Containing Rac1, IQGAP1, and Actin J. Biol. Chem., August 30, 2002; 277(36): 33081 - 33091. [Abstract] [Full Text] [PDF]
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D. L. Cioffi, T. M. Moore, J. Schaack, J. R. Creighton, D. M.F. Cooper, and T. Stevens Dominant regulation of interendothelial cell gap formation by calcium-inhibited type 6 adenylyl cyclase J. Cell Biol., June 24, 2002; 157(7): 1267 - 1278. [Abstract] [Full Text] [PDF]
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V. Vouret-Craviari, C. Bourcier, E. Boulter, and E. Van Obberghen-Schilling Distinct signals via Rho GTPases and Src drive shape changes by thrombin and sphingosine-1-phosphate in endothelial cells J. Cell Sci., June 15, 2002; 115(12): 2475 - 2484. [Abstract] [Full Text] [PDF]
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K. J. Bayless and G. E. Davis The Cdc42 and Rac1 GTPases are required for capillary lumen formation in three-dimensional extracellular matrices J. Cell Sci., March 15, 2002; 115(6): 1123 - 1136. [Abstract] [Full Text] [PDF]
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S. van Wetering, J. D. van Buul, S. Quik, F. P. J. Mul, E. C. Anthony, J.-P. t. Klooster, J. G. Collard, and P. L. Hordijk Reactive oxygen species mediate Rac-induced loss of cell-cell adhesion in primary human endothelial cells J. Cell Sci., January 5, 2002; 115(9): 1837 - 1846. [Abstract] [Full Text] [PDF]
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D. W. Lawrence, K. M. Comerford, and S. P. Colgan Role of VASP in reestablishment of epithelial tight junction assembly after Ca2+ switch Am J Physiol Cell Physiol, June 1, 2002; 282(6): C1235 - C1245. [Abstract] [Full Text] [PDF]
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