Novel role of microtubules in thrombin-induced endothelial barrier dysfunction

被引:163
作者
Birukova, AA
Birukov, KG
Smurova, K
Adyshev, D
Kaibuchi, K
Alieva, I
Garcia, JGN
Verin, AD
机构
[1] Johns Hopkins Univ, Sch Med, Div Pulm & Crit Care Med, Baltimore, MD 21224 USA
[2] Moscow MV Lomonosov State Univ, Belozersky Inst Physicochem Biol, Moscow 117234, Russia
[3] Nagoya Univ, Grad Sch Med, Dept Cell Pharmacol, Nagoya, Aichi, Japan
关键词
thrombin; G-proteins; Rho-kinase; tau; pulmonary endothelium;
D O I
10.1096/fj.04-2328com
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 Ser(409) and Ser(262) 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 Galpha12/13 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-induced 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.
引用
收藏
页码:1879 / 1890
页数:12
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