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JOURNAL ARTICLES
A unique pattern of expression of the four muscle regulatory factor proteins distinguishes somitic from embryonic, fetal and newborn mouse myogenic cells
T.H. Smith, N.E. Block, S.J. Rhodes, S.F. Konieczny, J.B. Miller
Development 1993 117: 1125-1133;
T.H. Smith
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N.E. Block
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S.J. Rhodes
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S.F. Konieczny
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J.B. Miller
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Summary

A unique pattern of expression of the four muscle regulatory factor (MRF) proteins was found to distinguish early somitic from embryonic, fetal and newborn limb myogenic cells in vitro. Expression of the myosin heavy chain (MHC), MyoD, myogenin, Myf-5, and MRF4 proteins was examined by immunocytochemistry in cultures of four distinct types of mouse myogenic cells: somitic (E8.5), embryonic (E11.5), fetal (E16.5) and newborn limb. In embryonic, fetal and newborn cultures, the MRF proteins were expressed in generally similar patterns: MyoD was the first MRF expressed; MyoD and myogenin were expressed by more cells than Myf-5 or MRF4; and each of the four MRFs was found both in cells that expressed MHC and in cells that did not express MHC. In cultures of somitic cells, in contrast, Myf-5 was expressed first and by more cells than MyoD or myogenin; MRF4 was not detected; and the MRFs were never found to be coexpressed with MHC in the same cell. Thus, some somitic cells had the unexpected ability to maintain MHC expression in the absence of detectable MRF protein expression. The different myogenic programs of embryonic, fetal and newborn myogenic cells are not, therefore, a simple result of qualitatively different MRF expression patterns, whereas myogenesis by somitic cells does include a unique pattern of MRF expression.

REFERENCES

    1. Bengal E.,
    2. Ransone L.,
    3. Scharfmann R.,
    4. Dwarki V.,
    5. Tapscott S.,
    6. Weintraub H.,
    7. Verma I.
    (1992) Functional antagonism between c-Jun and MyoD proteins: a direct physical association. Cell 68, 507–519
    OpenUrlCrossRefPubMedWeb of Science
    1. Blau H. M.,
    2. Pavlath G. K.,
    3. Hardeman E. C.,
    4. Chiu C.-P.,
    5. Silberstein L.,
    6. Webster S. G.,
    7. Miller S. C.,
    8. Webster C.
    (1985) Plasticity of the differentiated state. Science 230, 758–766
    OpenUrlAbstract/FREE Full Text
    1. Block N. E.,
    2. Miller J. B.
    (1992) Expression of MRF4, a myogenic helix-loop-helix protein, produces multiple changes in the myogenic program of BC3H-1 cells. Mol. Cell. Biol 12, 2484–2492
    OpenUrlAbstract/FREE Full Text
    1. Block N. E.,
    2. Menick D. R.,
    3. Robinson K. A.,
    4. Buse M. G.
    (1991) Effect of denervation on the expression of two glucose transporter isoforms in rat hindlimb muscle. J. Clin. Invest 88, 1546–1552
    1. Bober E.,
    2. Lyons G. E.,
    3. Braun T.,
    4. Cossu G.,
    5. Buckingham M. E.,
    6. Arnold H.-H.
    (1991) The muscle regulatory gene, myf-6, has a biphasic pattern of expression during early mouse development. J. Cell Biol 113, 1255–1266
    OpenUrlAbstract/FREE Full Text
    1. Braun T.,
    2. Buschhausen-Denker G.,
    3. Bober E.,
    4. Tannich E.,
    5. Arnold H.-H.
    (1989) A novel human muscle factor related to but distinct from MyoD1 induces myogenic conversion in 10T1/2 fibroblasts. EMBO J 8, 701–709
    OpenUrlPubMedWeb of Science
    1. Brennan T. J.,
    2. Edmondson D. G.,
    3. Olson E. N.
    (1990) Aberrant regulation of MyoD1 contributes to the partially defective myogenic phenotype of BC3H-1 cells. J. Cell Biol 110, 929–938
    OpenUrlAbstract/FREE Full Text
    1. Buonanno A.,
    2. Apone L.,
    3. Morasso M. I.,
    4. Beers R.,
    5. Brenner H. R.,
    6. Eftimie R.
    (1992) The MyoD family of myogenic factors is regulated by electrical activity: isolation and characterization of a mouse Myf-5 cDNA. Nucl. Acids Res 20, 539–544
    OpenUrlAbstract/FREE Full Text
    1. Chakraborty T.,
    2. Brennan T.,
    3. Olson E. N.
    (1991) Differential trans-activation of a muscle-specific enhancer by myogenic helix-loop-helix proteins is separable from DNA binding. J. Biol. Chem 266, 2878–2882
    OpenUrlAbstract/FREE Full Text
    1. Chen L.,
    2. Krause M.,
    3. Draper B.,
    4. Weintraub H.,
    5. Fire A.
    (1992) Body-wall muscle formation in Caenorhabditis elegans embryos that lack the MyoD homolog hlh-1. Science 256, 240–243
    OpenUrlAbstract/FREE Full Text
    1. Chomczynski P.,
    2. Sacchi N.
    (1987) A single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Analyt. Biochem 162, 156–159
    OpenUrlCrossRefPubMedWeb of Science
    1. Cossu G.,
    2. Molinaro M.
    (1987) Cell heterogeneity in the myogenic lineage. Curr. Top. Dev. Biol 23, 185–208
    OpenUrlCrossRefPubMedWeb of Science
    1. Crescenzi M.,
    2. Fleming T. P.,
    3. Lassar A. B.,
    4. Weintraub H.,
    5. Aaronson S. A.
    (1990) MyoD induces growth arrest independent of differentiation in normal and transformed cells. Proc. Natl. Acad. Sci. USA 87, 8442–8446
    OpenUrlAbstract/FREE Full Text
    1. Cusella-De Angelis M.,
    2. Lyons G.,
    3. Sonnin C.,
    4. De Angelis L.,
    5. Vivarelli E.,
    6. Farmer K.,
    7. Wright W.,
    8. Molinaro M.,
    9. Bouche M.,
    10. Buckingham M.,
    11. Cossu G.
    (1992) MyoD, myogenin independent differentiation of primordial myoblasts in mouse somites. J. Cell Biol 116, 1243–1255
    OpenUrlAbstract/FREE Full Text
    1. Davis R. L.,
    2. Weintraub H.,
    3. Lassar A. B.
    (1987) Expression of a single transfected cDNA converts fibroblasts to myoblasts. Cell 51, 987–1000
    OpenUrlCrossRefPubMedWeb of Science
    1. Edmondson D. G.,
    2. Brennan T. J.,
    3. Olson E. N.
    (1991) Mitogenic repression of myogenin autoregulation. J. Biol. Chem 266, 21343–21346
    OpenUrlAbstract/FREE Full Text
    1. Feinberg A. P.,
    2. Vogelstein B.
    (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Analyt. Biochem 132, 6–13
    OpenUrlCrossRefPubMedWeb of Science
    1. Fujisawa-Sehara A.,
    2. Nabeshima Y.,
    3. Komiya T.,
    4. Uetsuki T.,
    5. Asakura A.,
    6. Nabeshima N.
    (1992) Differential trans-activation of muscle-specific regulatory elements including the myosin light chain box by chicken MyoD, myogenin and MRF4. J. Biol. Chem 267, 10031–10038
    OpenUrlAbstract/FREE Full Text
    1. Gossett L.,
    2. Kelvin D.,
    3. Sternberg E.,
    4. Olson E. N.
    (1989) A new myocyte-specific enhancer binding factor that recognizes a conserved element associated with multiple muscle-specific genes. Mol. Cell. Biol 9, 5022–5033
    OpenUrlAbstract/FREE Full Text
    1. Hannon K.,
    2. Smith C. K., 2d.,
    3. Bales K. R.,
    4. Santerre R. F.
    (1992) Temporal and quantitative analysis of myogenic regulatory and growth factor gene expression in the developing mouse embryo. Dev. Biol 151, 137–144
    OpenUrlCrossRefPubMedWeb of Science
    1. Hinterberger T.,
    2. Sassoon D. A.,
    3. Rhodes S. J.,
    4. Konieczny S. F.
    (1991) Expression of the muscle regulatory factor MRF4 during somite and skeletal myofiber development. Dev. Biol 147, 144–156
    OpenUrlCrossRefPubMedWeb of Science
    1. Hopwood N. D.,
    2. Gurdon J. B.
    (1990) Activation of muscle genes without myogenesis by ectopic expression of MyoD in frog embryo cells. Nature 347, 197–200
    OpenUrlCrossRefPubMed
    1. Hopwood N. D.,
    2. Pluck A.,
    3. Gurdon J. B.
    (1991) Xenopus Myf-5 marks early muscle cells and can activate muscle genes ectopically in early embryos. Development 111, 551–560
    OpenUrlAbstract
    1. Lassar A. B.,
    2. Davis R. L.,
    3. Wright W. E.,
    4. Kadesch T.,
    5. Murre C.,
    6. Voronova A.,
    7. Baltimore D.,
    8. Weintraub H.
    (1991) Functional activity of myogenic HLH proteins requires hetero-oligomerization with E12/E47-like proteins in vivo. Cell 66, 305–315
    OpenUrlCrossRefPubMedWeb of Science
    1. Li L.,
    2. Chambard J. C.,
    3. Karin M.,
    4. Olson E. N.
    (1992) Fos and Jun repress transcriptional activation by myogenin and MyoD: the amino terminus of Jun can mediate repression. Genes Dev 6, 676–689
    OpenUrlAbstract/FREE Full Text
    1. Lin H.,
    2. Konieczny S. F.
    (1992) Identification of MRF4, myogenin and E12 oligomers by chemical cross-linking and two-dimensional gel electrophoresis. J. Biol. Chem 267, 4773–4780
    OpenUrlAbstract/FREE Full Text
    1. Mar J. H.,
    2. Ordahl C. P., Jr
    (1990) M-CAT binding factor, a novel trans-acting factor governing muscle-specific transcription. Mol. Cell. Biol 10, 4271–4283
    OpenUrlAbstract/FREE Full Text
    1. Martin J. F.,
    2. Li L.,
    3. Olson E. N.
    (1992) Repression of myogenin function by TGF-beta 1 is targeted at the basic helix-loop-helix motif and is independent of E2A products. J. Biol. Chem 267, 10956–10960
    OpenUrlAbstract/FREE Full Text
    1. Miller J. B.
    (1990) Myogenic programs of mouse muscle cell lines: Expression of myosin heavy chain isoforms, MyoD1 and myogenin. J. Cell Biol 111, 1149–1159
    OpenUrlAbstract/FREE Full Text
    1. Miller J. B.
    (1991) Myoblasts, myosins, MyoDs and the diversification of muscle fibers. Neuromusc. Disorders 1, 7–17
    OpenUrlCrossRefPubMed
    1. Miller J. B.
    (1992) Myoblast diversity in skeletal myogenesis: How much and to what end?. Cell 69, 1–3
    OpenUrlCrossRefPubMedWeb of Science
    1. Miller J. B.,
    2. Crow M. T.,
    3. Stockdale F. E.
    (1985) Slow and fast myosin heavy chain expression defines three types of myotubes in early muscle cell cultures. J. Cell Biol 101, 1643–1650
    OpenUrlAbstract/FREE Full Text
    1. Miller J. B.,
    2. Stockdale F. E.
    (1986) Developmental origins ofskeletal muscle fibers: Clonal analysis of myogenic cell lineages based on fast and slow myosin heavy chain expression. Proc. Nat. Acad. Sci. USA 83, 3860–3864
    OpenUrlAbstract/FREE Full Text
    1. Miller J. B.,
    2. Stockdale F. E.
    (1986) Developmental regulation of the multiple myogenic cell lineages of the avian embryo. J. Cell Biol 103, 2197–2208
    OpenUrlAbstract/FREE Full Text
    1. Miller J. B.,
    2. Stockdale F. E.
    (1987) What muscle cells know that nerves don't tell them. Trends Neurosci 10, 325–329
    OpenUrlCrossRef
    1. Miller J. B.,
    2. Teal S. B.,
    3. Stockdale F. E.
    (1989) Evolutionarily conserved sequences specific for striated muscle myosin heavy chain isoforms: Epitope mapping by cDNA expression. J. Biol. Chem 264, 13122–13130
    OpenUrlAbstract/FREE Full Text
    1. Miner J. H.,
    2. Wold B.
    (1990) Herculin, a fourth member of the MyoD family of myogenic regulatory genes. Proc. Nat. Acad. Sci. USA 87, 1089–1093
    OpenUrlAbstract/FREE Full Text
    1. Miner J. H.,
    2. Miller J. B.,
    3. Wold B.
    (1992) Skeletal muscle phenotypes initiated by ectopic MyoD in transgenic mouse heart. Development 114, 853–860
    OpenUrlAbstract
    1. Ordahl C. P.,
    2. LeDouarin N.
    (1992) Two myogenic lineages within the developing somite. Development 114, 339–353
    OpenUrlAbstract
    1. Pinney D.,
    2. Pearson-White S.,
    3. Konieczny S.,
    4. Latham K.,
    5. Emerson C.
    (1988) Myogenic lineage determination and differentiation: evidence for a regulatory gene pathway. Cell 53, 781–793
    OpenUrlCrossRefPubMedWeb of Science
    1. Reznikoff C. A.,
    2. Brankow D. W.,
    3. Heidelberger C.
    (1973) Establishment and characterization of a cloned line of C3H mouse embryo cells sensitive to postconfluence inhibition of division. Cancer Res 33, 3231–3238
    OpenUrlAbstract/FREE Full Text
    1. Rhodes S. J.,
    2. Konieczny S. F.
    (1989) Identification of MRF4: a new member of the muscle regulatory factor gene family. Genes Dev 3, 2050–2061
    OpenUrlAbstract/FREE Full Text
    1. Schubert D.,
    2. Harris J.,
    3. Devine C. E.,
    4. Heineman S.
    (1974) Characterization of a unique muscle cell line. J. Cell Biol 61, 398–413
    OpenUrlAbstract/FREE Full Text
    1. Smith D. B.,
    2. Johnson K. S.
    (1988) Single-step purification of polypeptides expressed in Escherichia coli as fusion with glutathione S-transferase. Gene 67, 31–40
    OpenUrlCrossRefPubMedWeb of Science
    1. Smith T. H.,
    2. Miller J. B.
    (1992) Distinct myogenic programs of embryonic and fetal mouse muscle cells: Expression of the perinatal myosin heavy chain isoform in vitro. Dev. Biol 149, 16–26
    OpenUrlCrossRefPubMedWeb of Science
    1. Sorrentino V.,
    2. Pepperkok R.,
    3. Davis R. L.,
    4. Ansorge W.,
    5. Phillipson L.
    (1990) Cell proliferation inhibited by MyoD1 independently of myogenic differentiation. Nature 345, 813–815
    OpenUrlCrossRefPubMed
    1. Stockdale F. E.
    (1990) The myogenic lineage: Evidence for multiple cellular precursors during avian limb development. Proc. Exp. Biol. Med 194, 71–75
    OpenUrlAbstract/FREE Full Text
    1. Sun X.-H.,
    2. Copeland N. G.,
    3. Jenkins N. A.,
    4. Baltimore D.
    (1991) Id proteins Id1 and Id2 selectively inhibit DNA binding by one class of helix-loop-helix proteins. Mol. Cell. Biol 11, 5603–5611
    OpenUrlAbstract/FREE Full Text
    1. Thompson W. R.,
    2. Nadal-Ginard B.,
    3. Mahdavi V.
    (1991) A MyoD1-independent muscle-specific enhancer controls the expression of the beta myosin heavy chain gene in skeletal and cardiac muscle cells. J. Biol. Chem 266, 22678–22688
    OpenUrlAbstract/FREE Full Text
    1. Vaidya T. B.,
    2. Rhodes S. J.,
    3. Taparowsky E. J.,
    4. Konieczny S. F.
    (1989) Fibroblast growth factor and transforming growth factorrepress transcription of the myogenic regulatory gene MyoD1. Mol. Cell. Biol 9, 3576–3579
    OpenUrlAbstract/FREE Full Text
    1. Weintraub H.,
    2. Davis R.,
    3. Tapscott S.,
    4. Thayer M.,
    5. Grause M.,
    6. Benezra R.,
    7. Blackwell T.,
    8. Turner D.,
    9. Rupp R.,
    10. Hollenberg S.,
    11. Zhuang Y.,
    12. Lassar A.
    (1991) The myoD gene family: Nodal point during specification of the muscle cell lineage. Science 251, 761–766
    OpenUrlAbstract/FREE Full Text
    1. Wright W. E.,
    2. Sassoon D. A.,
    3. Lin V. K.
    (1989) Myogenin, a factor regulating myogenesis, has a domain homologous to MyoD1. Cell 56, 607–617
    OpenUrlCrossRefPubMedWeb of Science
    1. Wright W. E.,
    2. Binder M.,
    3. Funk W.
    (1991) Cyclic amplification and target selection (CASTING). for the myogenin consensus binding site. Mol. Cell. Biol 11, 4104–4111
    OpenUrlAbstract/FREE Full Text
    1. Yutzey K. E.,
    2. Rhodes S. J.,
    3. Konieczny S. F.
    (1990) Differential trans activation associated with the muscle regulatory factors MyoD1, myogenin and MRF4. Mol. Cell. Biol 10, 3934–3944
    OpenUrlAbstract/FREE Full Text
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JOURNAL ARTICLES
A unique pattern of expression of the four muscle regulatory factor proteins distinguishes somitic from embryonic, fetal and newborn mouse myogenic cells
T.H. Smith, N.E. Block, S.J. Rhodes, S.F. Konieczny, J.B. Miller
Development 1993 117: 1125-1133;
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JOURNAL ARTICLES
A unique pattern of expression of the four muscle regulatory factor proteins distinguishes somitic from embryonic, fetal and newborn mouse myogenic cells
T.H. Smith, N.E. Block, S.J. Rhodes, S.F. Konieczny, J.B. Miller
Development 1993 117: 1125-1133;

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