Requirement of Rac activity for maintenance of capillary endothelial barrier properties

J. Waschke, W. Baumgartner, R. H. Adamson, M. Zeng, K. Aktories, H. Barth, C. Wilde, F. E. Curry, D. Drenckhahn

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Abstract

Our previous experiments indicated that GTPases, other than RhoA, are important for the maintenance of endothelial barrier integrity in both intact microvessels of rats and mice and cultured mouse myocardial endothelial (MyEnd) cell monolayers (J Physiol 539: 295-308, 2002). In the present study, we inhibited the endothelial GTPase Rac by Clostridium sordellii lethal toxin (LT) and investigated the relation between the degree of inhibition of Rac by glucosylation and increased endothelial barrier permeability. In rat venular microvessels, LT (200 ng/ml) increased hydraulic conductivity from a control value of 2.5 ± 0.6 to 100.8 ± 18.7 × 10-7 cm·s-1·cmH2O-1 after 80 min. In cultured My End cells exposed to LT (200 ng/ml), up to 60% of cellular Rac was glucosylated after 90 min, resulting in depolymerization of F-actin and interruptions of junctional distribution of vascular endothelial cadherin (VE-cadherin) and β-catenin as well as the formation of intercellular gaps. To understand the mechanism by which inhibition of Rac caused disassembly of adherens junctions, we used laser tweezers to quantify VE-cadherin-mediated adhesion. LT and cytochalasin D, an actin depolymerizing agent, both reduced adhesion of VE-cadherin-coated microbeads to the endothelial cell surface, whereas the inhibitor of Rho kinase Y-27632 did not. Stabilization of actin filaments by jasplakinolide completely blocked the effect of cytochalasin D but not of LT on bead adhesion. We conclude that Rac regulates endothelial barrier properties in vivo and in vitro by 1) modulation of actin filament polymerization and 2) acting directly on the tether between VE-cadherin and the cytoskeleton.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Heart and Circulatory Physiology
Volume286
Issue number1 55-1
StatePublished - Jan 2004

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Keywords

  • Actin
  • Adherens junction
  • Permeability
  • Rho proteins
  • Vascular endothelial cadherin

ASJC Scopus subject areas

  • Physiology

Cite this

Waschke, J., Baumgartner, W., Adamson, R. H., Zeng, M., Aktories, K., Barth, H., Wilde, C., Curry, F. E., & Drenckhahn, D. (2004). Requirement of Rac activity for maintenance of capillary endothelial barrier properties. American Journal of Physiology - Heart and Circulatory Physiology, 286(1 55-1).