Regulation of endothelial barrier function by the cAMP-dependent protein kinase

C. E. Patterson, H. Lum, K. L. Schaphorst, Alexander Dmitriyevich Verin, J. G.N. Garcia

Research output: Contribution to journalArticle

83 Citations (Scopus)

Abstract

Elevation of cAMP promotes the endothelial cell (EC) barrier and protects the lung from edema development. Thus, we tested the hypothesis that both increases and decreases in PKA modulate EC function and coordinate distribution of regulatory, adherence, and cytoskeletal proteins. Inhibition of PKA activity by RpcAMPS and activation by cholera toxin was verified by assay of kemptide phosphorylation in digitonin permeabilized EC. Inhibition of PKA by RpcAMPS or overexpression of the endogenous inhibitor, PKI, decreased monolayer electrical impedance and exacerbated the decreases produced by agonists (thrombin and PMA). RpcAMPS directly increased F-actin content and organization into stress fibers, increased co-staining of actin with both phosphatase 2B and myosin light chain kinase (MLCK), caused reorganization of focal adhesions, and decreased catenin at cell borders. These findings are similar to those evoked by thrombin. In contrast, cholera toxin prevented the agonist-induced resistance decrease and protein redistribution. Although PKA activation attenuated thrombin-induced myosin light chain (MLC) phosphorylation, PKA inhibition per se did not cause MLC phosphorylation or affect [Ca2+]i. These studies indicate that a decrease in PKA activity alone can produce disruption of barrier function via mechanisms not involving MLCK and support a central role for cAMP/PKA in regulation of cytoskeletal and adhesive protein function in EC which correlates with altered barrier function.

Original languageEnglish (US)
JournalEndothelium: Journal of Endothelial Cell Research
Volume7
Issue number4
DOIs
StatePublished - Jan 1 2000
Externally publishedYes

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Cyclic AMP-Dependent Protein Kinases
Endothelial Cells
Thrombin
Myosin-Light-Chain Kinase
Myosin Light Chains
Cytoskeletal Proteins
Cholera Toxin
Phosphorylation
kemptide
Actins
Digitonin
Catenins
Stress Fibers
Focal Adhesions
Electric Impedance
Phosphoric Monoester Hydrolases
Adhesives
Edema
Staining and Labeling
Lung

Keywords

  • Adherens junction
  • Cyclic AMP
  • Endothelium
  • Focal adhesion
  • Protein kinase A
  • Vascular permeability

ASJC Scopus subject areas

  • Physiology
  • Cell Biology

Cite this

Regulation of endothelial barrier function by the cAMP-dependent protein kinase. / Patterson, C. E.; Lum, H.; Schaphorst, K. L.; Verin, Alexander Dmitriyevich; Garcia, J. G.N.

In: Endothelium: Journal of Endothelial Cell Research, Vol. 7, No. 4, 01.01.2000.

Research output: Contribution to journalArticle

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