Abstract
Zinc finger-containing transcription factor GATA-4 and homeodomain Nkx-2.5 govern crucial developmental fates and have been found to promote cardiogenesis in embryonic cells exposed to the differentiating agent DMSO. Nevertheless, intracellular activators of these transcription factors are largely unknown. In this study, pluripotent P19 cells expressed the prodynorphin gene, an opioid gene encoding for the dynorphin family of opioid peptides. P19 cells were also able to synthesize and secrete dynorphin B, a biologically active end product of the prodynorphin gene. DMSO-primed GATA-4 and Nkx-2.5 gene expression was preceded by a marked increase in prodynorphin gene expression and dynorphin B synthesis and secretion. The DMSO effect occurred at the transcriptional level. In the absence of DMSO, dynorphin B triggered GATA-4 and Nkx-2.5 gene expression and led to the appearance of both alpha-myosin heavy chain and myosin light chain-2V transcripts, two markers of cardiac differentiation. Moreover, dynorphin B-exposed cells were positively stained in the presence of MF 20, a mouse monoclonal antibody raised against the alpha-myosin heavy chain. Opioid receptor antagonism and inhibition of opioid gene expression by a prodynorphin antisense phosphorothioate oligonucleotide blocked DMSO-induced cardiogenesis, suggesting an autocrine role of an opioid gene in developmental decisions.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Autocrine Communication
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Cell Differentiation / drug effects
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DNA-Binding Proteins / biosynthesis
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DNA-Binding Proteins / genetics
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DNA-Binding Proteins / physiology
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Dimethyl Sulfoxide / pharmacology
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Dynorphins / biosynthesis
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Dynorphins / genetics
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Dynorphins / pharmacology
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Endorphins / biosynthesis
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Endorphins / genetics
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Endorphins / pharmacology
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Enkephalins / biosynthesis
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Enkephalins / genetics*
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Enkephalins / physiology
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Fetal Heart / growth & development*
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Fetal Proteins / biosynthesis
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Fetal Proteins / genetics
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Fetal Proteins / physiology*
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GATA4 Transcription Factor
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Gene Expression Regulation, Developmental* / drug effects
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Homeobox Protein Nkx-2.5
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Homeodomain Proteins / biosynthesis
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Homeodomain Proteins / genetics
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Homeodomain Proteins / physiology
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Mice
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Oligonucleotides, Antisense / pharmacology
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Opioid Peptides / biosynthesis
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Opioid Peptides / genetics*
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Protein Precursors / biosynthesis
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Protein Precursors / genetics*
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Protein Precursors / physiology
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RNA, Messenger / biosynthesis
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RNA, Messenger / genetics
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Recombinant Fusion Proteins / biosynthesis
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Recombinant Fusion Proteins / genetics
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Stem Cells / cytology*
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Stem Cells / metabolism
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Thionucleotides / pharmacology
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Transcription Factors / biosynthesis
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Transcription Factors / genetics
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Transcription Factors / physiology
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Transfection
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Xenopus Proteins*
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Zinc Fingers / physiology
Substances
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DNA-Binding Proteins
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Endorphins
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Enkephalins
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Fetal Proteins
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GATA4 Transcription Factor
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Homeobox Protein Nkx-2.5
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Homeodomain Proteins
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NKX2-5 protein, human
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Nkx2-5 protein, mouse
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Oligonucleotides, Antisense
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Opioid Peptides
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Protein Precursors
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RNA, Messenger
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Recombinant Fusion Proteins
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Thionucleotides
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Transcription Factors
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Xenopus Proteins
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Dynorphins
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rimorphin
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preproenkephalin
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Dimethyl Sulfoxide