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Vascular Endothelial Growth Factor/Akt‐mediated endothelial Nitric Oxide Synthase Phosphorylation Regulates Endothelial Cell Division
Author(s) -
Gentile Carmine,
MuiseHelmericks Robin C.,
Drake Christopher J.
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.300.8
Subject(s) - enos , vasculogenesis , angiogenesis , microbiology and biotechnology , cd31 , pi3k/akt/mtor pathway , biology , cell division , protein kinase b , vascular endothelial growth factor , phosphorylation , nitric oxide , chemistry , signal transduction , endocrinology , nitric oxide synthase , cancer research , cell , biochemistry , stem cell , vegf receptors , progenitor cell
To evaluate the relevancy of nitric oxide (NO) to vasculogenesis mouse embryos were immunolabeled with antibodies that define the early endothelial cell (EC) lineage as well as antibodies that define the eNOS signaling pathway. Analysis revealed that isolated Flk1+/TAL1+/CD31−/VE‐Cadherin− cells (angioblasts) were eNOS− whereas primitive ECs (PECs), Flk1+/TAL1+/CD31+/VE‐Cadherin− cells and definitive ECs Flk1+/TAL1−/CD31+/VE‐Cadherin+ cells were eNOS+. Although the majority of the eNOS+ cells were phospho‐eNOS (PeNOS)−, PECs adjacent to the dorsal aortae and allantoic PECs and ECs were PeNOS+. Analysis of cultured 8.5dpc allantoides revealed a correlation between P‐eNOS expression and EC division. A correlation between PeNOS expression and EC division was also observed in nocodazole‐treated HUVECs. To evaluate VEGF as a regulator of eNOS phosphorylation, analysis of HUVECs treated with either VEGF alone or VEGF plus L‐NAME (an inhibitor of NO production) revealed that all dividing cells in VEGF treated cultures were PeNOS+ and that L‐NAME blocked VEFG‐mediated EC division. Our findings suggest that VEGF acting through PI3K/Akt‐mediated eNOS phosphorylation regulates PEC/EC division. Current studies seek to determine if PeNOS acts to regulate PEC/EC division via the nitrosylation of cyclins and to determine whether this pathway is applicable to angiogenesis. Grant Funding Source : NIH RO1 HL080168 (CJD)