Opioid peptide gene expression primes cardiogenesis in embryonal pluripotent stem cells.
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.
Department of Biomedical Sciences, Division of Biochemistry, Laboratory of Cardiovascular Research, University of Sassari, Sassari, Italy. chim_med@ssmain.uniss.it
C
C
Ventura
M
M
Maioli
D000818:Animals; D019898:Autocrine Communication; D002454:Cell Differentiation / Q000187:drug effects; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D004399:Dynorphins / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology; D004723:Endorphins / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology; D004745:Enkephalins / Q000096:biosynthesis / Q000235:genetics* / Q000502:physiology; D005318:Fetal Heart / Q000254:growth & development*; D005326:Fetal Proteins / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D050991:GATA4 Transcription Factor; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D000072576:Homeobox Protein Nkx-2.5; D018398:Homeodomain Proteins / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology; D051379:Mice; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D018847:Opioid Peptides / Q000096:biosynthesis / Q000235:genetics*; D011498:Protein Precursors / Q000096:biosynthesis / Q000235:genetics* / Q000502:physiology; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D013873:Thionucleotides / Q000494:pharmacology; D014157:Transcription Factors / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology; D014162:Transfection; D029867:Xenopus Proteins; D016335:Zinc Fingers / Q000502:physiology
D004268:DNA-Binding Proteins; D004723:Endorphins; D004745:Enkephalins; D005326:Fetal Proteins; D050991:GATA4 Transcription Factor; D000072576:Homeobox Protein Nkx-2.5; D018398:Homeodomain Proteins; C095552:NKX2-5 protein, human; C484469:Nkx2-5 protein, mouse; D016376:Oligonucleotides, Antisense; D018847:Opioid Peptides; D011498:Protein Precursors; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D013873:Thionucleotides; D014157:Transcription Factors; D029867:Xenopus Proteins; D004399:Dynorphins; C036200:rimorphin; C033660:preproenkephalin; D004121:Dimethyl Sulfoxide