title: VEGF-E activates endothelial nitric oxide synthase to induce angiogenesis via cGMP and PKG-independent pathways.
abstract: Vascular endothelial growth factor-A (VEGF), which binds to both VEGF receptor-1 (Flt1) and VEGFR-2 (KDR/Flk-1), requires nitric oxide (NO) to induce angiogenesis in a cGMP-dependent manner. Here we show that VEGF-E, a VEGFR-2-selective ligand stimulates NO release and tube formation in human umbilical vein endothelial cells (HUVEC). Inhibition of phospholipase Cgamma (PLCgamma) with U73122 abrogated VEGF-E induced endothelial cell migration, tube formation and NO release. Inhibition of endothelial nitric oxide synthase (eNOS) using l-NNA blocked VEGF-E-induced NO release and angiogenesis. Pre-incubation of HUVEC with the soluble guanylate cyclase inhibitor, ODQ, or the protein kinase G (PKG) inhibitor, KT-5823, had no effect on angiogenesis suggesting that the action of VEGF-E is cGMP-independent. Our data provide the first demonstration that VEGFR-2-mediated NO signaling and subsequent angiogenesis is through a mechanism that is dependent on PLCgamma but independent of cGMP and PKG.
authors.affiliation: Department of Reproductive and Vascular Biology, Centre for Cardiovascular Sciences, Institute of Biomedical Research, The Medical School, University of Birmingham, Edgbaston, UK.
authors.forename: Melissa
authors.initials: M
authors.lastname: Cudmore
authors.forename: Shakil
authors.initials: S
authors.lastname: Ahmad
authors.forename: Bahjat
authors.initials: B
authors.lastname: Al-Ani
authors.forename: Peter
authors.initials: P
authors.lastname: Hewett
authors.forename: Suborna
authors.initials: S
authors.lastname: Ahmed
authors.forename: Asif
authors.initials: A
authors.lastname: Ahmed
mesh_terms: D000818:Animals; D002227:Carbazoles / Q000494:pharmacology; D002465:Cell Movement / Q000187:drug effects; D002478:Cells, Cultured; D006152:Cyclic GMP / Q000378:metabolism*; D017869:Cyclic GMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004963:Estrenes / Q000494:pharmacology; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D018919:Neovascularization, Physiologic / Q000187:drug effects; D009569:Nitric Oxide / Q000378:metabolism; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D019335:Nitroarginine / Q000494:pharmacology; D051966:Phospholipase C gamma / Q000037:antagonists & inhibitors / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D011760:Pyrrolidinones / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects*; D013552:Swine; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000378:metabolism; D014764:Viral Proteins / Q000494:pharmacology*
chemical_list: D002227:Carbazoles; D004791:Enzyme Inhibitors; D004963:Estrenes; D007211:Indoles; D011760:Pyrrolidinones; C084758:VEGF-like protein, Orf virus; D014764:Viral Proteins; C060229:1-(6-((3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione; C073601:KT 5823; D019335:Nitroarginine; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase; D040301:Vascular Endothelial Growth Factor Receptor-2; D017869:Cyclic GMP-Dependent Protein Kinases; D051966:Phospholipase C gamma; D006152:Cyclic GMP