title: Adenosine monophosphate-activated kinase alpha1 promotes endothelial barrier repair.
abstract: The vascular endothelium responds to damage through activation of multiple signaling events that restore cell-cell adhesion and vascular integrity. However, the molecular mechanisms that integrate these events are not clearly defined. Herein, we identify a previously unexpected role for adenosine monophosphate-activated protein kinase (AMPK) in pulmonary microvascular endothelial cell (PMVEC) repair. PMVECs selectively express the AMPKα1 catalytic subunit, pharmacological and short hairpin RNA-mediated inhibition of which attenuates Ca(2+) entry in these cells induced by the inflammatory Ca(2+)-signaling mimetic thapsigargin. We find that AMPKα1 activity is required for the formation of PMVEC cell-cell networks in a prorepair environment and for monolayer resealing after wounding. Decreasing AMPKα1 expression reduces barrier resistance in PMVEC monolayers, results consistent with a role for AMPKα1 in cell-cell adhesion. AMPKα1 colocalizes and coimmunoprecipitates with the adherens junction protein N-cadherin and cofractionates with proteins selectively expressed in caveolar membranes. Assessment of permeability, by measuring the filtration coefficient (K(f)) in isolated perfused lungs, confirmed that AMPK activation contributes to barrier repair in vivo. Our findings thus provide novel evidence for AMPKα1 in Ca(2+) influx-mediated signaling and wound repair in the endothelium.
authors.affiliation: Department of Anesthesiology, University of Alabama, Birmingham, Alabama, USA.
authors.forename: Judy
authors.initials: J
authors.lastname: Creighton
authors.forename: MingYuan
authors.initials: M
authors.lastname: Jian
authors.forename: Sarah
authors.initials: S
authors.lastname: Sayner
authors.forename: Mikhail
authors.initials: M
authors.lastname: Alexeyev
authors.forename: Paul A
authors.initials: PA
authors.lastname: Insel
mesh_terms: D055372:AMP-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D015820:Cadherins; D002118:Calcium / Q000378:metabolism; D020013:Calcium Signaling; D051242:Caveolin 1 / Q000378:metabolism; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000201:enzymology / Q000378:metabolism* / Q000473:pathology; D004730:Endothelium, Vascular / Q000166:cytology / Q000293:injuries / Q000502:physiology; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D008297:Male; D021381:Protein Transport; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D015820:Cadherins; D051242:Caveolin 1; D055372:AMP-Activated Protein Kinases; C568305:Prkaa1 protein, rat; D002118:Calcium