Novel role of microtubules in thrombin-induced endothelial barrier dysfunction

Anna A. Birukova, Konstantin G. Birukov, Ksenya Smurova, Djanybek Adyshev, Kozo Kaibuchi, Irina Alieva, Joe G.N. Garcia, Alexander Dmitriyevich Verin

Research output: Contribution to journalArticle

163 Scopus citations

Abstract

Disturbances in endothelial cell (EC) barrier regulation are critically dependent upon rearrangements of EC actin cytoskeleton. However, the role of microtubule (MT) network in the regulation of EC permeability is not well understood. We examined involvement of MT remodeling in thrombin-induced EC permeability and explored MT regulation by heterotrimeric G12/13 proteins and by small GTPase Rho. Thrombin induced phosphorylation of MT regulatory protein tau at Ser409 and Ser262 and peripheral MT disassembly, which was linked to increased EC permeability. MT stabilization by taxol attenuated thrombin-induced permeability, actin remodeling, and paracellular gap formation and diminished thrombin-induced activation of Rho and Rho-kinase. Expression of activated Ga 12/13 subunits involved in thrombin-mediated signaling or their effector p115RhoGEF involved in Rho activation caused MT disassembly, whereas p115RhoGEF-specific negative regulator RGS preserved MT from thrombin-hiduced disassembly. Consistent with these results, expression of activated RhoA and Rho-kinase induced MT disassembly. Conversely, thrombin-induced disassembly of peripheral MT network was attenuated by expression of dominant negative RhoA and Rho-kinase mutants or by pharmacological inhibition of Rho-kinase. Collectively, our data demonstrate for the first time a critical involvement of MT disassembly in thrombin-induced EC barrier dysfunction and indicate G-protein-dependent mechanisms of thrombin-induced MT alteration.

Original languageEnglish (US)
Pages (from-to)1879-1890
Number of pages12
JournalFASEB Journal
Volume18
Issue number15
DOIs
StatePublished - Dec 1 2004
Externally publishedYes

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Keywords

  • G-proteins
  • Pulmonary endothelium
  • Rho-kinase
  • Tau
  • Thrombin

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Genetics

Cite this

Birukova, A. A., Birukov, K. G., Smurova, K., Adyshev, D., Kaibuchi, K., Alieva, I., Garcia, J. G. N., & Verin, A. D. (2004). Novel role of microtubules in thrombin-induced endothelial barrier dysfunction. FASEB Journal, 18(15), 1879-1890. https://doi.org/10.1096/fj.04-2328com