title: Caveolin-1 scaffolding domain promotes leukocyte adhesion by reduced basal endothelial nitric oxide-mediated ICAM-1 phosphorylation in rat mesenteric venules.
abstract: Exogenously applied caveolin-1 scaffolding domain (CAV) has been shown to inhibit inflammatory mediator-induced nitric oxide (NO) production and NO-mediated increases in microvessel permeability. However, the effect of CAV on endothelial basal NO that prevents leukocyte adhesion remains unknown. This study aims to investigate the roles of exogenously applied CAV in endothelial basal NO production, leukocyte adhesion, and adhesion-induced changes in microvessel permeability. Experiments were conducted in individually perfused rat mesenteric venules. Microvessel permeability was determined by measuring hydraulic conductivity (Lp). NO was quantified with fluorescence imaging in DAF-2-loaded vessels. Perfusing venules with CAV inhibited basal NO production without affecting basal Lp. Resuming blood flow in CAV-perfused vessels significantly increased leukocyte adhesion. The firmly adherent leukocytes altered neither basal Lp nor adherens junction integrity. Increases in Lp occurred only upon formyl-Met-Leu-Phe application that induces release of reactive oxygen species from the adherent leukocytes. The application of NO synthase inhibitor showed similar results to CAV, and NO donor abolished CAV-mediated leukocyte adhesion. Immunofluorescence staining showed increases in binding of ICAM-1 to an adhesion-blocking antibody concurrent with a Src-dependent ICAM-1 phosphorylation following CAV perfusion. Pre-perfusing vessels with anti-ICAM-1 blocking antibody or a Src kinase inhibitor attenuated CAV-induced leukocyte adhesion. These results indicate that the application of CAV, in addition to preventing excessive NO-mediated permeability increases, also causes reduction of basal NO and promotes ICAM-1-mediated leukocyte adhesion through Src activation-mediated ICAM-1 phosphorylation. CAV-induced leukocyte adhesion was uncoupled from leukocyte oxidative burst and microvessel barrier function, unless in the presence of a secondary stimulation.
authors.affiliation: Department of Physiology and Pharmacology, School of Medicine, West Virginia University, Morgantown, West Virginia.
authors.forename: Sulei
authors.initials: S
authors.lastname: Xu
authors.forename: Xueping
authors.initials: X
authors.lastname: Zhou
authors.forename: Dong
authors.initials: D
authors.lastname: Yuan
authors.forename: Yanchun
authors.initials: Y
authors.lastname: Xu
authors.forename: Pingnian
authors.initials: P
authors.lastname: He
mesh_terms: D000818:Animals; D051943:Antennapedia Homeodomain Protein / Q000494:pharmacology; D002199:Capillary Permeability / Q000187:drug effects; D051242:Caveolin 1 / Q000494:pharmacology*; D002448:Cell Adhesion / Q000187:drug effects*; D004730:Endothelium, Vascular / Q000187:drug effects* / Q000201:enzymology / Q000276:immunology; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005260:Female; D005455:Fluorescent Antibody Technique; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism*; D007962:Leukocytes / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D008643:Mesentery / Q000098:blood supply*; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009569:Nitric Oxide / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III / Q000037:antagonists & inhibitors* / Q000378:metabolism; D010446:Peptide Fragments / Q000494:pharmacology*; D010766:Phosphorylation; D011485:Protein Binding; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D017382:Reactive Oxygen Species / Q000378:metabolism; D013997:Time Factors; D014699:Venules / Q000187:drug effects / Q000201:enzymology / Q000276:immunology; D019061:src-Family Kinases / Q000378:metabolism
chemical_list: D051943:Antennapedia Homeodomain Protein; D051242:Caveolin 1; D004791:Enzyme Inhibitors; D010446:Peptide Fragments; D047428:Protein Kinase Inhibitors; D017382:Reactive Oxygen Species; C571371:caveolin-1 (82-101); D018799:Intercellular Adhesion Molecule-1; D009569:Nitric Oxide; D009240:N-Formylmethionine Leucyl-Phenylalanine; D052250:Nitric Oxide Synthase Type III; C496316:Nos3 protein, rat; D019061:src-Family Kinases
keywords: ICAM-1 phorsphorylation; caveolin-1; leukocyte adhesion; microvessel permeability; reactive oxygen species
references.citation: Am J Physiol. 1993 Nov;265(5 Pt 2):H1623-8
references.pmid: 7902014
references.citation: Am J Physiol Heart Circ Physiol. 2004 Jan;286(1):H195-201
references.pmid: 12946927
references.citation: Anesthesiology. 1997 Sep;87(3):591-8
references.pmid: 9316965
references.citation: Nat Rev Immunol. 2007 Oct;7(10):803-15
references.pmid: 17893694
references.citation: Am J Physiol Cell Physiol. 2002 Dec;283(6):C1745-51
references.pmid: 12388068
references.citation: Nat Rev Immunol. 2007 Sep;7(9):678-89
references.pmid: 17717539
references.citation: Proc Natl Acad Sci U S A. 1991 Jun 1;88(11):4651-5
references.pmid: 1675786
references.citation: Circ Res. 2003 May 30;92(10):1089-97
references.pmid: 12714560
references.citation: Circulation. 1992 Oct;86(4):1302-9
references.pmid: 1394936
references.citation: Exp Physiol. 1995 May;80(3):359-72
references.pmid: 7640005
references.citation: Am J Physiol Heart Circ Physiol. 2005 Jun;288(6):H2869-77
references.pmid: 15665052
references.citation: J Biol Chem. 1997 Jul 25;272(30):18522-5
references.pmid: 9228013
references.citation: Am J Physiol Heart Circ Physiol. 2002 Dec;283(6):H2420-30
references.pmid: 12388263
references.citation: Cell Metab. 2009 Jul;10(1):48-54
references.pmid: 19583953
references.citation: Am J Physiol. 1996 Dec;271(6 Pt 2):H2735-9
references.pmid: 8997338
references.citation: Am J Physiol Heart Circ Physiol. 2000 Oct;279(4):H2017-23
references.pmid: 11009493
references.citation: Am J Physiol. 1994 Jun;266(6 Pt 2):H2369-73
references.pmid: 7912900
references.citation: Am J Physiol. 1999 Jun;276(6):H1943-50
references.pmid: 10362674
references.citation: Nat Med. 2000 Dec;6(12):1362-7
references.pmid: 11100121
references.citation: Am J Physiol Heart Circ Physiol. 2005 Mar;288(3):H1331-8
references.pmid: 15498822
references.citation: Am J Physiol Heart Circ Physiol. 2011 Jul;301(1):H108-14
references.pmid: 21536843
references.citation: Free Radic Biol Med. 2012 Feb 1;52(3):556-592
references.pmid: 22154653
references.citation: J Biol Chem. 1997 Oct 10;272(41):25437-40
references.pmid: 9325253
references.citation: Am J Physiol. 1997 Jan;272(1 Pt 2):H176-85
references.pmid: 9038936
references.citation: Am J Physiol. 1996 Dec;271(6 Pt 2):H2377-87
references.pmid: 8997296
references.citation: Arterioscler Thromb Vasc Biol. 2004 Jan;24(1):98-105
references.pmid: 14563650
references.citation: J Biol Chem. 1996 Nov 15;271(46):29182-90
references.pmid: 8910575
references.citation: Inflammation. 1992 Aug;16(4):295-305
references.pmid: 1526662
references.citation: Am J Physiol Heart Circ Physiol. 2006 Jan;290(1):H365-72
references.pmid: 16155097
references.citation: J Cell Biol. 1992 Nov;119(4):935-44
references.pmid: 1385447
references.citation: Lancet. 1987 Nov 7;2(8567):1057-8
references.pmid: 2889967
references.citation: Am J Pathol. 2010 Aug;177(2):998-1003
references.pmid: 20581061
references.citation: Arterioscler Thromb Vasc Biol. 2011 Jun;31(6):1342-50
references.pmid: 21474822
references.citation: Am J Physiol Heart Circ Physiol. 2011 Nov;301(5):H1788-97
references.pmid: 21873500
references.citation: Proc Natl Acad Sci U S A. 2005 Jan 18;102(3):904-8
references.pmid: 15640348
references.citation: Am J Physiol Cell Physiol. 2004 May;286(5):C1195-202
references.pmid: 15075219
references.citation: J Vasc Surg. 1996 Aug;24(2):187-93
references.pmid: 8752027
references.citation: Cardiovasc Res. 2010 Jul 15;87(2):281-90
references.pmid: 20472564
references.citation: Circ Res. 1993 Jul;73(1):164-71
references.pmid: 7685251
references.citation: Cardiovasc Res. 2010 Jul 15;87(2):340-7
references.pmid: 20080986
references.citation: Trends Pharmacol Sci. 2003 Feb;24(2):91-5
references.pmid: 12559774
references.citation: Cancer Cell. 2003 Jul;4(1):31-9
references.pmid: 12892711
references.citation: Circ Res. 1994 Jun;74(6):1133-40
references.pmid: 7910528
references.citation: Cardiovasc Res. 2010 Jul 15;87(2):254-61
references.pmid: 20462865
references.citation: Am J Physiol. 1992 Feb;262(2 Pt 2):H611-5
references.pmid: 1539722
references.citation: Circ Res. 1995 Jan;76(1):30-9
references.pmid: 7528112
references.citation: Proc Natl Acad Sci U S A. 2005 Jan 18;102(3):761-6
references.pmid: 15637154
references.citation: Am J Physiol. 1995 Jun;268(6 Pt 2):H2224-31
references.pmid: 7541959
references.citation: J Immunol. 2004 Aug 1;173(3):2068-77
references.pmid: 15265942