Inhibition of precardiac mesoderm cell proliferation by antisense oligodeoxynucleotide complementary to fibroblast growth factor-2 (FGF-2).
This laboratory recently reported the occurrence of concentrated deposits of fibroblast growth factor-2 (FGF-2; basic FGF)-like proteins in the sarcoplasm of embryonic cardiac myocytes from the earliest stage of avian heart development (M. H. Parlow, D. L. Bolender, N. P. Kokan-Moore, and J. Lough, Dev. Biol. 146, 139, 1991). To determine the role, if any, of FGF-2 during embryonic cardiogenesis, the proliferative and functional effects of treating cultured anterior lateral plate mesoderm from Hamburger-Hamilton stage 6 embryos with antisense oligodeoxynucleotide (ODN) complementary to FGF-2 mRNA were determined. Within 2 days of culture in defined medium, the explanted monolayer normally differentiates into a multilayer of proliferative cells that express sarcomeric alpha-actin and exhibit rhythmic contractions. The inclusion of 25 microM ODN that is complementary to the second exon of chicken FGF-2 mRNA caused a 50% inhibition in these cells' proliferative ability as judged by incorporation of 5'-bromodeoxyuridine; contractility was similarly inhibited. These effects were prevented by including recombinant human FGF-2 protein in the medium. Treatment with sense ODN did not cause inhibition. Inhibition of FGF-2 protein synthesis in the explanted tissue by antisense ODN was verified by immunoprecipitation analysis. These results point to a critical role for FGF-2 in the autocrine regulation of proliferation, and perhaps differentiative function, of embryonic cardiac myocytes.
Sugi
Y
Y
Department of Cellular Biology and Anatomy, Medical College of Wisconsin, Milwaukee 53226.
Sasse
J
J
Lough
J
J
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D001483:Base Sequence; D002455:Cell Division / Q000187:drug effects; D002642:Chick Embryo; D002645:Chickens; D005091:Exons; D016222:Fibroblast Growth Factor 2 / Q000032:analysis / Q000235:genetics*; D006321:Heart / Q000196:embryology*; D008648:Mesoderm / Q000166:cytology* / Q000187:drug effects / Q000502:physiology; D008969:Molecular Sequence Data; D009200:Myocardial Contraction / Q000187:drug effects; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D009924:Organ Culture Techniques; D010455:Peptides / Q000138:chemical synthesis / Q000276:immunology; D016133:Polymerase Chain Reaction / Q000379:methods; D012333:RNA, Messenger / Q000235:genetics
D000906:Antibodies; D016376:Oligonucleotides, Antisense; D010455:Peptides; D012333:RNA, Messenger; D016222:Fibroblast Growth Factor 2