Resealing of endothelial junctions by focal adhesion kinase.
Endothelial cell (EC) junctions determine vascular barrier properties and are subject to transient opening to allow liquid flux from blood to tissue. Although EC junctions open in the presence of permeability-enhancing factors, including oxidants, the mechanisms by which they reseal remain inadequately understood. To model opening and resealing of EC junctions in the presence of an oxidant, we quantified changes in H(2)O(2)-induced transendothelial resistance (TER) in monolayers of rat lung microvascular EC. During a 30-min exposure, H(2)O(2) (100 microM) decreased TER for an initial approximately 10 min, indicating junctional opening. Subsequently, despite continuous presence of H(2)O(2), TER recovered to baseline, indicating the activation of junctional resealing mechanisms. These bimodal TER transients matched the time course of loss and then gain of E-cadherin at EC junctions. The timing of the TER decrease matched the onset of focal adhesion formation, while F-actin increase at the cell periphery occurred with a time course that complemented the recovery of peripheral E-cadherin. In monolayers expressing a focal adhesion kinase (FAK) mutant (del-FAK) that inhibits FAK activity, the initial H(2)O(2)-induced junctional opening was present, although the subsequent junctional recovery was blocked. Expression of transfected E-cadherin was evident at the cell periphery of wild-type but not del-FAK-expressing EC. E-cadherin overexpression in del-FAK-expressing EC failed to effect major rescue of the junctional resealing response. These findings indicate that in oxidant-induced EC junction opening, FAK plays a critical role in remodeling the adherens junction to reseal the barrier.
Quadri
Sadiqa K
SK
Lung Biology Laboratory, College of Physicians and Surgeons, Columbia University, St. Luke's-Roosevelt Hospital Center, 432 West 58th Street, New York, NY 10019, USA. skq1@columbia.edu
Bhattacharya
Jahar
J
D022005:Adherens Junctions / Q000187:drug effects / Q000201:enzymology* / Q000648:ultrastructure; D000818:Animals; D015820:Cadherins / Q000378:metabolism; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000187:drug effects / Q000201:enzymology* / Q000648:ultrastructure; D051417:Focal Adhesion Kinase 1 / Q000172:deficiency / Q000235:genetics / Q000502:physiology*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000633:toxicity; D008564:Membrane Potentials / Q000187:drug effects; D016877:Oxidants / Q000633:toxicity; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects
D015820:Cadherins; D016877:Oxidants; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D006861:Hydrogen Peroxide; D051417:Focal Adhesion Kinase 1; C496052:Ptk2 protein, rat