Abstract
Molecular mechanisms controlling arterial-venous specification have not been fully elucidated. Previously, we established an embryonic stem cell differentiation system and demonstrated that activation of cAMP signaling together with VEGF induces arterial endothelial cells (ECs) from Flk1(+) vascular progenitor cells. Here, we show novel arterial specification machinery regulated by Notch and beta-catenin signaling. Notch and GSK3beta-mediated beta-catenin signaling were activated downstream of cAMP through phosphatidylinositol-3 kinase. Forced activation of Notch and beta-catenin with VEGF completely reconstituted cAMP-elicited arterial EC induction, and synergistically enhanced target gene promoter activity in vitro and arterial gene expression during in vivo angiogenesis. A protein complex with RBP-J, the intracellular domain of Notch, and beta-catenin was formed on RBP-J binding sites of arterial genes in arterial, but not venous ECs. This molecular machinery for arterial specification leads to an integrated and more comprehensive understanding of vascular signaling.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Arteries / cytology
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Arteries / embryology*
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Arteries / metabolism
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Biomarkers / metabolism
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Cell Differentiation / physiology
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Cyclic AMP / metabolism
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Endothelial Cells / cytology
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Endothelial Cells / physiology*
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Glycogen Synthase Kinase 3 / genetics
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Glycogen Synthase Kinase 3 / metabolism
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Glycogen Synthase Kinase 3 beta
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Immunoglobulin J Recombination Signal Sequence-Binding Protein / genetics
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Immunoglobulin J Recombination Signal Sequence-Binding Protein / metabolism
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Mice
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Neovascularization, Physiologic / physiology
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Phosphatidylinositol 3-Kinases / genetics
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Phosphatidylinositol 3-Kinases / metabolism
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Phosphoinositide-3 Kinase Inhibitors
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Receptors, Notch / genetics
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Receptors, Notch / metabolism*
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Signal Transduction / physiology*
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Stem Cells / cytology
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Stem Cells / physiology*
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Vascular Endothelial Growth Factor A / metabolism
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Vascular Endothelial Growth Factor Receptor-2 / genetics
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Vascular Endothelial Growth Factor Receptor-2 / metabolism
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Veins / cytology
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Veins / embryology
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Veins / metabolism
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beta Catenin / genetics
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beta Catenin / metabolism*
Substances
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Biomarkers
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Immunoglobulin J Recombination Signal Sequence-Binding Protein
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Phosphoinositide-3 Kinase Inhibitors
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Rbpj protein, mouse
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Receptors, Notch
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Vascular Endothelial Growth Factor A
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beta Catenin
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Cyclic AMP
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Vascular Endothelial Growth Factor Receptor-2
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Glycogen Synthase Kinase 3 beta
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Gsk3b protein, mouse
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Glycogen Synthase Kinase 3