eNOS-induced vascular barrier disruption in retinopathy by c-Src activation and tyrosine phosphorylation of VE-cadherin

Elife. 2021 Apr 28;10:e64944. doi: 10.7554/eLife.64944.


Background: Hypoxia and consequent production of vascular endothelial growth factor A (VEGFA) promote blood vessel leakiness and edema in ocular diseases. Anti-VEGFA therapeutics may aggravate hypoxia; therefore, therapy development is needed.

Methods: Oxygen-induced retinopathy was used as a model to test the role of nitric oxide (NO) in pathological neovascularization and vessel permeability. Suppression of NO formation was achieved chemically using L-NMMA, or genetically, in endothelial NO synthase serine to alanine (S1176A) mutant mice.

Results: Suppression of NO formation resulted in reduced retinal neoangiogenesis. Remaining vascular tufts exhibited reduced vascular leakage through stabilized endothelial adherens junctions, manifested as reduced phosphorylation of vascular endothelial (VE)-cadherin Y685 in a c-Src-dependent manner. Treatment with a single dose of L-NMMA in established retinopathy restored the vascular barrier and prevented leakage.

Conclusions: We conclude that NO destabilizes adheren junctions, resulting in vascular hyperpermeability, by converging with the VEGFA/VEGFR2/c-Src/VE-cadherin pathway.

Funding: This study was supported by the Swedish Cancer foundation (19 0119 Pj ), the Swedish Research Council (2020-01349), the Knut and Alice Wallenberg foundation (KAW 2020.0057) and a Fondation Leducq Transatlantic Network of Excellence Grant in Neurovascular Disease (17 CVD 03). KAW also supported LCW with a Wallenberg Scholar grant (2015.0275). WCS was supported by Grants R35 HL139945, P01 HL1070205, AHA MERIT Award. DV was supported by grants from the Deutsche Forschungsgemeinschaft, SFB1450, B03, and CRU342, P2.

Keywords: Src; VE-cadherin; cell biology; medicine; mouse; nitric oxide; retinal vasculature; retinopathy of prematurity; vascular barrier.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adherens Junctions / genetics
  • Adherens Junctions / metabolism
  • Amino Acid Motifs
  • Animals
  • Antigens, CD / chemistry*
  • Antigens, CD / genetics
  • Antigens, CD / metabolism*
  • CSK Tyrosine-Protein Kinase / genetics
  • CSK Tyrosine-Protein Kinase / metabolism*
  • Cadherins / chemistry*
  • Cadherins / genetics
  • Cadherins / metabolism*
  • Capillary Permeability
  • Endothelial Cells / enzymology
  • Endothelial Cells / metabolism
  • Endothelium, Vascular / enzymology
  • Endothelium, Vascular / metabolism
  • Humans
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Neovascularization, Pathologic
  • Nitric Oxide / metabolism
  • Nitric Oxide Synthase Type III / genetics
  • Nitric Oxide Synthase Type III / metabolism*
  • Phosphorylation
  • Retinal Diseases / enzymology*
  • Retinal Diseases / genetics
  • Retinal Diseases / metabolism
  • Tyrosine / metabolism*
  • Vascular Endothelial Growth Factor A / genetics
  • Vascular Endothelial Growth Factor A / metabolism


  • Antigens, CD
  • Cadherins
  • Vascular Endothelial Growth Factor A
  • cadherin 5
  • Nitric Oxide
  • Tyrosine
  • Nitric Oxide Synthase Type III
  • Nos3 protein, mouse
  • CSK Tyrosine-Protein Kinase

Associated data

  • Dryad/10.5061/dryad.x69p8czhv