Abstract
Hypoxia Inducible Factor (HIF), consisting of HIF1alpha and ARNT (HIF1beta) subunits, activates multiple genes in response to oxygen (O(2)) deprivation. Arnt(-/-) mice exhibit substantial defects in blood cell and vessel development. We demonstrate that hypoxia accelerates the expression of Brachyury (a mesoderm-specific transcription factor), BMP4 (a mesoderm-promoting growth factor) and FLK1 (a marker of hemangioblasts, the bipotential progenitor of endothelial and hematopoietic cells) in differentiating ES cell cultures. Significantly, proliferation of embryonic hemangioblasts (BL-CFCs) is regulated by hypoxia, as Arnt(+/+) ES cells generate increased numbers of FLK1(+) cells, and BL-CFCs with accelerated kinetics in response to low O(2). This response is HIF-dependent as Arnt(-/-) ES cells produce fewer FLK1(+) cells and BL-CFCs, under both normoxic and hypoxic conditions. Interestingly, this defect is rescued when Arnt(-/-) ES cells are co-cultured with Arnt(+/+) ES cells. Vegf(+/-)or Vegf(-/-) ES cells generate proper numbers of FLK1(+) cells but fewer BL-CFCs, suggesting that additional factors regulated by HIF (other than VEGF) are involved in these early events. Thus, hypoxic responses are important for the establishment of various progenitor cells, including early mesoderm and its differentiation into hemangioblasts. Together these data suggest that ineffective responses to hypoxia in Arnt(-/-) embryos abrogate proper cardiovascular development during early embryogenesis, including the pathways controlling hemangioblast differentiation.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Animals
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Aryl Hydrocarbon Receptor Nuclear Translocator
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Bone Morphogenetic Protein 4
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Bone Morphogenetic Proteins / genetics
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Cell Count
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Cell Differentiation*
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Cell Division
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Cells, Cultured
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Coculture Techniques
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DNA-Binding Proteins / deficiency
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DNA-Binding Proteins / genetics
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DNA-Binding Proteins / metabolism
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Fetal Proteins / genetics
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Gene Deletion
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Growth Substances / pharmacology
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Hypoxia / genetics
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Hypoxia / physiopathology*
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Hypoxia-Inducible Factor 1
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Hypoxia-Inducible Factor 1, alpha Subunit
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Kinetics
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Mesoderm / cytology*
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Mesoderm / metabolism*
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Mice
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Mice, Knockout
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Nuclear Proteins / genetics
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Phenotype
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RNA, Messenger / genetics
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RNA, Messenger / metabolism
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Receptors, Aryl Hydrocarbon / deficiency
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Receptors, Aryl Hydrocarbon / genetics
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Receptors, Aryl Hydrocarbon / metabolism
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Stem Cells / cytology
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Stem Cells / metabolism
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T-Box Domain Proteins / genetics
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Transcription Factors / deficiency
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Transcription Factors / genetics
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Transcription Factors / metabolism
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Vascular Endothelial Growth Factor A / deficiency
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Vascular Endothelial Growth Factor A / genetics
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Vascular Endothelial Growth Factor Receptor-2 / genetics
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Vascular Endothelial Growth Factor Receptor-2 / metabolism
Substances
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Arnt protein, mouse
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Bmp4 protein, mouse
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Bone Morphogenetic Protein 4
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Bone Morphogenetic Proteins
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DNA-Binding Proteins
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Fetal Proteins
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Growth Substances
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Hif1a protein, mouse
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Hypoxia-Inducible Factor 1
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Hypoxia-Inducible Factor 1, alpha Subunit
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Nuclear Proteins
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RNA, Messenger
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Receptors, Aryl Hydrocarbon
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T-Box Domain Proteins
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Transcription Factors
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Vascular Endothelial Growth Factor A
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Aryl Hydrocarbon Receptor Nuclear Translocator
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Vascular Endothelial Growth Factor Receptor-2
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Brachyury protein