Protein kinase A attenuates endothelial cell barrier dysfunction induced by microtubule disassembly

Anna A. Birukova, Feng Liu, Joe G.N. Garcia, Alexander Dmitriyevich Verin

Research output: Contribution to journalArticle

61 Citations (Scopus)

Abstract

Cross talk between the actin cytoskeleton and the microtubule (MT) network plays a critical role in regulation of endothelial permeability. We have previously demonstrated that MT disruption by nocodazole results in increases in MLC phosphorylation, actomyosin contraction, cell retraction, and paracellular gap formation, cardinal features of endothelial barrier dysfunction (Verin AD, Birukova A, Wang P, Liu F, Becker P, Birukov K, and Garcia JG. Am J Physiol Lung Cell Mol Physiol 281: L565-L574, 2001: Birukova AA, Smurova K, Birukov KG, Usatyuk P, Liu F, Kaibuchi K, Ricks-Cord A, Natarajan V, Alieva A, Garcia JG, and Verin AD. J Cell Physiol. In press.). Although activation of PKA opposes barrier-disrupting effects of edemagenic agents on confluent EC monolayers, information about the molecular mechanisms of PKA-mediated EC barrier protection is limited. Our results suggest that MT disassembly alters neither intracellular cAMP levels nor PKA enzymatic activity; however, elevation of cAMP levels and PKA activation by either cholera toxin or forskolin dramatically attenuates the decline in transendothelial electrical resistance induced by nocodazole in human pulmonary EC. Barrier-protective effects of PKA on EC were associated with PKA-mediated inhibition of nocodazole-induced stress fiber formation. Rho activation, phosphorylation of myosin phosphatase regulatory subunit at Thr696. and decreased MLC phosphorylation. In addition, forskolin pretreatment attenuated MT disassembly induced by nocodazole. These results suggest a critical role for PKA activity in stabilization of MT cytoskeleton and provide a novel mechanism for cAMP-mediated regulation of Rho-induced actin cytoskeletal remodeling, actomyosin contraction, and EC barrier dysfunction induced by MT disassembly.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Lung Cellular and Molecular Physiology
Volume287
Issue number1 31-1
DOIs
StatePublished - Jul 1 2004
Externally publishedYes

Fingerprint

Cyclic AMP-Dependent Protein Kinases
Microtubules
Nocodazole
Endothelial Cells
Actomyosin
Phosphorylation
Colforsin
Myosin-Light-Chain Phosphatase
Lung
Stress Fibers
Cholera Toxin
Cytoskeleton
Electric Impedance
Actin Cytoskeleton
Actins
Permeability

Keywords

  • Actin
  • Myosin
  • Myosin phosphatase
  • Phosphorylation
  • Pulmonary endothelium
  • Rho-associated kinase
  • RhoA

ASJC Scopus subject areas

  • Physiology
  • Pulmonary and Respiratory Medicine
  • Physiology (medical)
  • Cell Biology

Cite this

Protein kinase A attenuates endothelial cell barrier dysfunction induced by microtubule disassembly. / Birukova, Anna A.; Liu, Feng; Garcia, Joe G.N.; Verin, Alexander Dmitriyevich.

In: American Journal of Physiology - Lung Cellular and Molecular Physiology, Vol. 287, No. 1 31-1, 01.07.2004.

Research output: Contribution to journalArticle

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