Role of cadherin internalization in hydrogen peroxide-mediated endothelial permeability.
Exposure of endothelial monolayers to hydrogen peroxide results in increased solute permeability in a time- and dose-dependent fashion. This effect is prevented by either staurosporine, an inhibitor of PKC, or by Gö6976, an inhibitor of "classical" PKC isoforms. Immunohistochemistry of peroxide-treated monolayers illustrates a loss of cadherin staining at cell junctions and gap formation predominantly at tri-cellular junctions. Both staurosporine and Gö6976 prevented peroxide-induced gap formation. Peroxide also stimulated internalization of cadherins as measured by the trypsin protection assay, which was not blocked by staurosporine or Gö6976. These data suggest that peroxide causes: 1) a time- and dose-dependent increase in permeability and dose-dependent increase in gap formation, both of which are PKC dependent; and 2) promotes PKC-independent cadherin internalization. These data indicate that cadherin internalization may be part of the mechanism through which oxidants regulate solute permeability.
Kevil
C G
CG
LSU Medical Center, Department of Molecular and Cellular Physiology, Shreveport, LA 71130, USA.
Ohno
N
N
Gute
D C
DC
Okayama
N
N
Robinson
S A
SA
Chaney
E
E
Alexander
J S
JS
D000818:Animals; D015820:Cadherins / Q000378:metabolism* / Q000502:physiology; D002199:Capillary Permeability / Q000187:drug effects*; D002227:Carbazoles / Q000494:pharmacology; D002417:Cattle; D002478:Cells, Cultured; D003513:Cycloheximide / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000166:cytology / Q000201:enzymology / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D007211:Indoles / Q000494:pharmacology; D007365:Intercellular Junctions / Q000187:drug effects; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism; D011651:Pulmonary Artery
D015820:Cadherins; D002227:Carbazoles; D007211:Indoles; C081021:Go 6976; D003513:Cycloheximide; D006861:Hydrogen Peroxide; D011493:Protein Kinase C