Abstract
Embryonic stem (ES) cell pluripotency and differentiation are controlled by a network of transcription factors and signaling molecules. Transcription factors such as Oct4 and Nanog are required for self-renewal and maintain the undifferentiated state of ES cells. Decreases in the expression of these factors indicate the initiation of differentiation of ES cells. Inactivation of the gene encoding the orphan nuclear receptor GCNF showed that it plays an important role in the repression of Oct4 expression in somatic cells during early embryonic development. GCNF-/- ES cells were isolated to study the function of GCNF in the down-regulation of pluripotency genes during differentiation. Loss of repression of ES cell marker genes Oct4, Nanog, Sox2, FGF4, and Stella was observed upon treatment of GCNF-/- ES cells with retinoic acid. The loss of repression of pluripotency genes is either a direct or indirect consequence of loss of GCNF. Both the Oct4 and Nanog genes are direct targets of GCNF repression during ES cell differentiation and early mouse embryonic development. In contrast Sox2 and FGF4 are indirectly regulated by GCNF through Oct4. These findings establish a central role for GCNF in the repression of pluripotency gene expression during retinoic acid-induced ES cell differentiation.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Animals
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Blotting, Northern
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Blotting, Western
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Cell Differentiation
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Cell Line
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Cell Nucleus / metabolism
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Chromatin Immunoprecipitation
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DNA-Binding Proteins / metabolism
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DNA-Binding Proteins / physiology*
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Down-Regulation
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Embryo, Mammalian / cytology*
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Female
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Fibroblast Growth Factor 4 / metabolism
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Genotype
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Homeodomain Proteins / metabolism
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In Situ Hybridization
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Male
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Mice
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Mice, Transgenic
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Microscopy, Fluorescence
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Models, Genetic
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Nanog Homeobox Protein
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Nuclear Receptor Subfamily 6, Group A, Member 1
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Octamer Transcription Factor-3 / metabolism
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Phenotype
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Plasmids / metabolism
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Protein Binding
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Receptors, Cytoplasmic and Nuclear / metabolism
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Receptors, Cytoplasmic and Nuclear / physiology*
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Response Elements
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Reverse Transcriptase Polymerase Chain Reaction
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SOXB1 Transcription Factors
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Signal Transduction
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Stem Cells / cytology*
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Time Factors
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Trans-Activators / metabolism
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Transfection
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Tretinoin / metabolism
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Tretinoin / pharmacology*
Substances
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DNA-Binding Proteins
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FGF4 protein, human
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Fibroblast Growth Factor 4
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Homeodomain Proteins
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NR6A1 protein, human
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Nanog Homeobox Protein
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Nanog protein, mouse
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Nr6a1 protein, mouse
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Nuclear Receptor Subfamily 6, Group A, Member 1
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Octamer Transcription Factor-3
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Pou5f1 protein, mouse
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Receptors, Cytoplasmic and Nuclear
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SOXB1 Transcription Factors
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Sox2 protein, mouse
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Trans-Activators
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Tretinoin