TY - JOUR
T1 - Involvement of microtubules and Rho pathway in TGF-β1-induced lung vascular barrier dysfunction
AU - Birukova, Anna A.
AU - Birukov, Konstantin G.
AU - Adyshev, Djanibek
AU - Usatyuk, Peter
AU - Natarajan, Viswanathan
AU - Garcia, Joe G.N.
AU - Verin, Alexander D.
PY - 2005/9
Y1 - 2005/9
N2 - Transforming growth factor-β1 (TGF-β1) is a cytokine critically involved in acute lung injury and endothelial cell (EC) barrier dysfunction. We have studied TGF-β1-mediated signaling pathways and examined a role of microtubule (MT) dynamics in TGF-β1-induced actin cytoskeletal remodeling and EC barrier dysfunction. TGF-β1 (0.1-50 ng/ml) induced dose-dependent decrease in transendothelial electrical resistance (TER) in bovine pulmonary ECs, which was linked to increased actin stress fiber formation, myosin light chain (MLC) phosphorylation, EC retraction, and gap formation. Inhibitor of TGF-β1 receptor kinase RI (5 μM) abolished TGF-β1-induced TER decline, whereas inhibitor of caspase-3 zVAD (10 μM) was without effect. TGF-β1-induced EC barrier dysfunction was linked to partial dissolution of peripheral MT meshwork and decreased levels of stable (acetylated) MT pool, whereas MT stabilization by taxol (5 μM) attenuated TGF-β1-induced barrier dysfunction and actin remodeling. TGF-β1 induced sustained activation of small GTPase Rho and its effector Rho-kinase; phosphorylation of myosin binding subunit of myosin specific phosphatase; MLC phosphorylation; EC contraction; and gap formation, which was abolished by inhibition of Rho and Rho-kinase, and by MT stabilization with taxol. Finally, elevation of intracellular cAMP induced by forskolin (50 μM) attenuated TGF-β1-induced barrier dysfunction, MLC phosphorylation, and protected the MT peripheral network. These results suggest a novel role for MT dynamics in the TGF-β1-mediated Rho regulation, EC barrier dysfunction, and actin remodeling.
AB - Transforming growth factor-β1 (TGF-β1) is a cytokine critically involved in acute lung injury and endothelial cell (EC) barrier dysfunction. We have studied TGF-β1-mediated signaling pathways and examined a role of microtubule (MT) dynamics in TGF-β1-induced actin cytoskeletal remodeling and EC barrier dysfunction. TGF-β1 (0.1-50 ng/ml) induced dose-dependent decrease in transendothelial electrical resistance (TER) in bovine pulmonary ECs, which was linked to increased actin stress fiber formation, myosin light chain (MLC) phosphorylation, EC retraction, and gap formation. Inhibitor of TGF-β1 receptor kinase RI (5 μM) abolished TGF-β1-induced TER decline, whereas inhibitor of caspase-3 zVAD (10 μM) was without effect. TGF-β1-induced EC barrier dysfunction was linked to partial dissolution of peripheral MT meshwork and decreased levels of stable (acetylated) MT pool, whereas MT stabilization by taxol (5 μM) attenuated TGF-β1-induced barrier dysfunction and actin remodeling. TGF-β1 induced sustained activation of small GTPase Rho and its effector Rho-kinase; phosphorylation of myosin binding subunit of myosin specific phosphatase; MLC phosphorylation; EC contraction; and gap formation, which was abolished by inhibition of Rho and Rho-kinase, and by MT stabilization with taxol. Finally, elevation of intracellular cAMP induced by forskolin (50 μM) attenuated TGF-β1-induced barrier dysfunction, MLC phosphorylation, and protected the MT peripheral network. These results suggest a novel role for MT dynamics in the TGF-β1-mediated Rho regulation, EC barrier dysfunction, and actin remodeling.
UR - http://www.scopus.com/inward/record.url?scp=21844450693&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=21844450693&partnerID=8YFLogxK
U2 - 10.1002/jcp.20359
DO - 10.1002/jcp.20359
M3 - Article
C2 - 15828024
AN - SCOPUS:21844450693
SN - 0021-9541
VL - 204
SP - 934
EP - 947
JO - Journal of Cellular Physiology
JF - Journal of Cellular Physiology
IS - 3
ER -