title: Shear stress promotes nitric oxide production in endothelial cells by sub-cellular delocalization of eNOS: A basis for shear stress mediated angiogenesis.
abstract: This study aims to investigate the role of shear stress in cellular remodeling and angiogenesis with relation to nitric oxide (NO). We observed a 2-fold increase in endothelial cell (EC) migration in relation to actin re-arrangements under 15 dyne/cm(2) shear stress. Blocking NO production inhibited the migration and ring formation of ECs by 6-fold and 5-fold, respectively under shear stress. eNOS-siRNA knockdown technique also ascertained a 3-fold reduction in shear stress mediated ring formation. In ovo artery ligation model with a half and complete flow block for 30 min showed a reduction of angiogenesis by 50% and 70%, respectively. External stimulation with NO donor showed a 2-fold recovery in angiogenesis under both half and complete flow block conditions. NO intensity clustering studies by using Diaminofluorescein diacetate (DAF-2DA) probed endothelial monolayer depicted pattern-changes in NO distribution and cluster formation of ECs under shear stress. Immunofluorescence and live cell studies revealed an altered sub-cellular localization pattern of eNOS and phospho-eNOS under shear stress. In conclusion, shear-induced angiogenesis is mediated by nitric oxide dependent EC migration.
authors.affiliation: Vascular Biology Lab, AU-KBC Research Centre, Anna University, Chennai, India.
authors.forename: Gopi Krishna
authors.initials: GK
authors.lastname: Kolluru
authors.forename: Swaraj
authors.initials: S
authors.lastname: Sinha
authors.forename: Syamantak
authors.initials: S
authors.lastname: Majumder
authors.forename: Ajit
authors.initials: A
authors.lastname: Muley
authors.forename: Jamila H
authors.initials: JH
authors.lastname: Siamwala
authors.forename: Ravi
authors.initials: R
authors.lastname: Gupta
authors.forename: Suvro
authors.initials: S
authors.lastname: Chatterjee
mesh_terms: D000818:Animals; D002642:Chick Embryo; D042783:Endothelial Cells / Q000201:enzymology* / Q000378:metabolism / Q000648:ultrastructure; D045462:Endothelium-Dependent Relaxing Factors / Q000378:metabolism; D006801:Humans; D018919:Neovascularization, Physiologic; D009569:Nitric Oxide / Q000096:biosynthesis*; D052250:Nitric Oxide Synthase Type III / Q000032:analysis* / Q000378:metabolism*; D010766:Phosphorylation; D013314:Stress, Mechanical
chemical_list: D045462:Endothelium-Dependent Relaxing Factors; D009569:Nitric Oxide; D052250:Nitric Oxide Synthase Type III