VEGF-mediated phosphorylation of eNOS regulates angioblast and embryonic endothelial cell proliferation

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Abstract

To evaluate potential roles of nitric oxide (NO) in the regulation of the endothelial lineage and neovascular processes (vasculogenesis and angiogenesis) we evaluated endothelial nitric oxide synthase (eNOS) and phosphorylated eNOS (p-eNOS) expression in 7.2-8.5 days post-coitum (dpc) mouse embryos. Analysis revealed that p-eNOS(S1177) but not P-eNOS(S617) or P-eNOS(T495) was expressed in a subpopulation of angioblasts (TAL-1+/Flk-1+/CD31-/CD34-/VE-Cadherin-) at 7.2 dpc. A role of the VEGF/Akt1/eNOS signaling pathway in the regulation of the endothelial cell (EC) lineage was suggested by the strong correlation observed between cell division and p-eNOS(S1177) expression in both angioblasts and embryonic endothelial cells (EECs, TAL-1+/Flk-1+/CD31+/CD34+/VE-Cadherin+). Our studies using Akt1 null mouse embryos show a reduction in p-eNOS(S1177) expression in angioblast and EECs that is correlated with a decrease in endothelial cell proliferation and results in changes in VEGF-induced vascular patterning. Further, we show that VEGF-mediated cell proliferation in Flk-1+ cells in allantoic cultures is decreased by pharmacological inhibitors of the VEGF/Akt1/eNOS signaling pathways. Taken together, our findings suggest that VEGF-mediated eNOS phosphorylation on Ser1177 regulates angioblast and EEC division, which underlies the formation of blood vessels and vascular networks. © 2012 Elsevier Inc.

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Gentile, C., Muise-Helmericks, R. C., & Drake, C. J. (2013). VEGF-mediated phosphorylation of eNOS regulates angioblast and embryonic endothelial cell proliferation. Developmental Biology, 373(1), 163–175. https://doi.org/10.1016/j.ydbio.2012.10.020

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