title: eNOS-derived nitric oxide regulates endothelial barrier function through VE-cadherin and Rho GTPases.
abstract: Transient disruption of endothelial adherens junctions and cytoskeletal remodeling are responsible for increases in vascular permeability induced by inflammatory stimuli and vascular endothelial growth factor (VEGF). Nitric oxide (NO) produced by endothelial NO synthase (eNOS) is crucial for VEGF-induced changes in permeability in vivo; however, the molecular mechanism by which endogenous NO modulates endothelial permeability is not clear. Here, we show that the lack of eNOS reduces VEGF-induced permeability, an effect mediated by enhanced activation of the Rac GTPase and stabilization of cortical actin. The loss of NO increased the recruitment of the Rac guanine-nucleotide-exchange factor (GEF) TIAM1 to adherens junctions and VE-cadherin (also known as cadherin 5), and reduced Rho activation and stress fiber formation. In addition, NO deficiency reduced VEGF-induced VE-cadherin phosphorylation and impaired the localization, but not the activation, of c-Src to cell junctions. The physiological role of eNOS activation is clear given that VEGF-, histamine- and inflammation-induced vascular permeability is reduced in mice bearing a non-phosphorylatable knock-in mutation of the key eNOS phosphorylation site S1176. Thus, NO is crucial for Rho GTPase-dependent regulation of cytoskeletal architecture leading to reversible changes in vascular permeability.
authors.affiliation: Department of Pharmacology and Vascular Biology and Therapeutics Program, Yale University School of Medicine, New Haven, CT 06520, USA.
authors.forename: Annarita
authors.initials: A
authors.lastname: Di Lorenzo
authors.forename: Michelle I
authors.initials: MI
authors.lastname: Lin
authors.forename: Takahisa
authors.initials: T
authors.lastname: Murata
authors.forename: Shira
authors.initials: S
authors.lastname: Landskroner-Eiger
authors.forename: Michael
authors.initials: M
authors.lastname: Schleicher
authors.forename: Milankumar
authors.initials: M
authors.lastname: Kothiya
authors.forename: Yasuko
authors.initials: Y
authors.lastname: Iwakiri
authors.forename: Jun
authors.initials: J
authors.lastname: Yu
authors.forename: Paul L
authors.initials: PL
authors.lastname: Huang
authors.forename: William C
authors.initials: WC
authors.lastname: Sessa
mesh_terms: D022005:Adherens Junctions / Q000378:metabolism; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism*; D000081247:CSK Tyrosine-Protein Kinase; D015820:Cadherins / Q000378:metabolism*; D002199:Capillary Permeability; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000201:enzymology; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism*; D020662:Guanine Nucleotide Exchange Factors / Q000378:metabolism*; D006801:Humans; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009569:Nitric Oxide / Q000502:physiology*; D052250:Nitric Oxide Synthase Type III / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D021381:Protein Transport; D022502:Stress Fibers / Q000378:metabolism; D000075684:T-Lymphoma Invasion and Metastasis-inducing Protein 1; D042461:Vascular Endothelial Growth Factor A / Q000502:physiology; D019061:src-Family Kinases / Q000378:metabolism; D000099152:Cadherin 5
chemical_list: D015703:Antigens, CD; D000081247:CSK Tyrosine-Protein Kinase; D015820:Cadherins; D020662:Guanine Nucleotide Exchange Factors; D009569:Nitric Oxide; D052250:Nitric Oxide Synthase Type III; D000075684:T-Lymphoma Invasion and Metastasis-inducing Protein 1; D042461:Vascular Endothelial Growth Factor A; D019061:src-Family Kinases; D000099152:Cadherin 5; C087334:TIAM1 protein, human; C496314:NOS3 protein, human; C000631346:CSK protein, human
keywords: Cadherin 5; Cytoskeleton; Nitric oxide; Src; VE-cadherin; VEGF; eNOS
references.citation: Am J Physiol Heart Circ Physiol. 2004 Oct;287(4):H1590-8
references.pmid: 15155260
references.citation: Science. 1998 Jun 26;280(5372):2074-5
references.pmid: 9669963
references.citation: Cell. 1992 Aug 7;70(3):401-10
references.pmid: 1643658
references.citation: Nat Med. 2003 Jun;9(6):669-76
references.pmid: 12778165
references.citation: J Appl Physiol (1985). 2003 Mar;94(3):1193-203
references.pmid: 12482769
references.citation: Vascul Pharmacol. 2002 Nov;39(4-5):187-99
references.pmid: 12747959
references.citation: Mol Cell. 2010 Aug 13;39(3):468-76
references.pmid: 20705246
references.citation: Proc Natl Acad Sci U S A. 2005 Jan 18;102(3):761-6
references.pmid: 15637154
references.citation: J Cell Sci. 1997 Sep;110 ( Pt 17):2065-77
references.pmid: 9378757
references.citation: J Physiol. 2006 Jul 1;574(Pt 1):275-81
references.pmid: 16675496
references.citation: J Cell Biol. 2004 Oct 25;167(2):223-9
references.pmid: 15504909
references.citation: Am J Physiol Lung Cell Mol Physiol. 2006 Dec;291(6):L1232-45
references.pmid: 16891393
references.citation: J Biol Chem. 1996 Jun 7;271(23):13556-60
references.pmid: 8662891
references.citation: J Biol Chem. 2006 Nov 24;281(47):35593-7
references.pmid: 17005550
references.citation: J Biol Chem. 1996 Aug 23;271(34):20246-9
references.pmid: 8702756
references.citation: Exp Cell Res. 2001 Sep 10;269(1):73-87
references.pmid: 11525641
references.citation: Microvasc Res. 2001 Nov;62(3):252-62
references.pmid: 11678628
references.citation: Nat Rev Mol Cell Biol. 2004 Apr;5(4):261-70
references.pmid: 15071551
references.citation: J Biol Chem. 2010 Feb 5;285(6):3806-3814
references.pmid: 19948721
references.citation: Arterioscler Thromb Vasc Biol. 1999 Oct;19(10):2286-97
references.pmid: 10521356
references.citation: Proc Natl Acad Sci U S A. 2009 Aug 25;106(34):14552-7
references.pmid: 19622728
references.citation: Oncogene. 2000 Jul 27;19(32):3632-41
references.pmid: 10951569
references.citation: J Cell Biol. 1999 Nov 15;147(4):831-44
references.pmid: 10562284
references.citation: Circ Res. 2002 May 3;90(8):904-10
references.pmid: 11988492
references.citation: Science. 1996 Jul 12;273(5272):245-8
references.pmid: 8662509
references.citation: Nat Med. 2000 Dec;6(12):1362-7
references.pmid: 11100121
references.citation: J Cell Biol. 2011 May 30;193(5):841-50
references.pmid: 21624953
references.citation: Cell. 1992 Aug 7;70(3):389-99
references.pmid: 1643657
references.citation: J Clin Invest. 2005 Aug;115(8):2119-27
references.pmid: 16075056
references.citation: Microcirculation. 1999 Mar;6(1):23-44
references.pmid: 10100187
references.citation: Trends Cell Biol. 2001 Dec;11(12):471-7
references.pmid: 11719051
references.citation: Mol Biol Cell. 2002 Apr;13(4):1175-89
references.pmid: 11950930
references.citation: J Exp Med. 2011 Nov 21;208(12):2393-401
references.pmid: 22025303
references.citation: J Clin Invest. 1998 Jun 1;101(11):2567-78
references.pmid: 9616228
references.citation: Oncogene. 1993 Jul;8(7):1875-86
references.pmid: 8510932
references.citation: Circ Res. 2012 Aug 17;111(5):553-63
references.pmid: 22777005
references.citation: J Clin Invest. 1997 Dec 15;100(12):3131-9
references.pmid: 9399960
references.citation: Cardiovasc Res. 2001 Feb 16;49(3):568-81
references.pmid: 11166270
references.citation: Science. 1997 Nov 21;278(5342):1464-6
references.pmid: 9367959
references.citation: J Cell Biol. 1996 Dec;135(6 Pt 1):1551-64
references.pmid: 8978822
references.citation: Science. 1998 Jan 23;279(5350):509-14
references.pmid: 9438836
references.citation: J Cell Sci. 1998 Jul;111 ( Pt 13):1853-65
references.pmid: 9625748
references.citation: J Clin Invest. 2001 Sep;108(5):689-701
references.pmid: 11544274
references.citation: Nat Med. 2003 Mar;9(3):357-62
references.pmid: 12598894
references.citation: Cell. 2004 Jan 23;116(2):167-79
references.pmid: 14744429
references.citation: Mol Cell. 1999 Dec;4(6):915-24
references.pmid: 10635317
references.citation: FASEB J. 2003 Dec;17(15):2240-9
references.pmid: 14656986
references.citation: J Cell Biol. 1998 Nov 30;143(5):1385-98
references.pmid: 9832565
references.citation: J Cell Sci. 2004 May 15;117(Pt 12):2427-9
references.pmid: 15159447
references.citation: Sci Signal. 2009 Aug 04;2(82):ra41
references.pmid: 19654415
references.citation: Am J Physiol Lung Cell Mol Physiol. 2001 Sep;281(3):L565-74
references.pmid: 11504682
references.citation: Am J Physiol Lung Cell Mol Physiol. 2002 Jun;282(6):L1330-8
references.pmid: 12003790
references.citation: Exp Cell Res. 1999 Nov 1;252(2):401-15
references.pmid: 10527630
references.citation: J Biol Chem. 1999 May 7;274(19):13198-204
references.pmid: 10224076
references.citation: J Cell Biol. 1999 Nov 29;147(5):1009-22
references.pmid: 10579721
references.citation: Cancer Cell. 2003 Jul;4(1):31-9
references.pmid: 12892711
references.citation: J Cell Sci. 2000 Feb;113 ( Pt 3):471-82
references.pmid: 10639334
references.citation: Arterioscler Thromb Vasc Biol. 2003 Feb 1;23(2):211-7
references.pmid: 12588761
references.citation: J Clin Invest. 1997 Jun 1;99(11):2625-34
references.pmid: 9169492
references.citation: Am J Pathol. 1997 Feb;150(2):713-25
references.pmid: 9033284
references.citation: Proc Natl Acad Sci U S A. 2001 Feb 27;98(5):2604-9
references.pmid: 11226286
references.citation: Am J Physiol. 1999 May;276(5):C1148-53
references.pmid: 10329964