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eNOS/NO and their role in modifying the vascular barrier in retinopathy - Source data

Citation

Love, Dominic; Ninchoji, Takeshi; Claesson-Welsh, Lena (2021), eNOS/NO and their role in modifying the vascular barrier in retinopathy - Source data, Dryad, Dataset, https://doi.org/10.5061/dryad.x69p8czhv

Abstract

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 (eNOS) 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, indicated by 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.

Conclusion: We conclude that NO destabilizes adherens 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 01 H), 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.  

Methods

The data uploaded here contains excel work sheets.

Figure (1-5)_Ninchoji et al_ 16-11-2020-RA-eLife-64944_source data.xlsx - Each figure and it accompanying supplemental figure are on a individual spread sheets (labelled with the figure number). The data for each specific figure is labelled on said sheets. The data contained within is raw data obtained from various experiments/methods and reflects what is shown in the figures.

Supplementary file 1A-C.xlsx - The tables contained in this spread sheet contain the weights of each mouse used in each experiment. Each table corresponds with the label in the article text.