Summary
The activation of mononuclear muscle precursor cells after crush injury to mouse tibialis anterior muscles was monitored in vivo by in situ hybridization with MyoD1 and myogenin probes. These genes are early markers of skeletal muscle differentiation and have been extensively studied in vitro. The role in vivo of these regulatory proteins during myogenesis of mature muscle has not been studied previously. MyoD1 and myogenin mRNA were present in occasional mononuclear cells of uninjured muscle. Increased MyoD1 and myogenin mRNA sequences in mononuclear cells were detected as early as 6 h after injury, peaked between 24 and 48 h, and thereafter declined to pre-injury levels at about 8 days. The mRNAs were detected in mononuclear cells throughout the muscle, with the majority of cells located some distance from the site of crush injury. The presence of MyoD1 and myogenin mRNA at 6 to 48 h indicates that transcription of these genes is occurring at the same time as replication of muscle precursor cells in vivo. At no time were significant levels of mRNA for these genes detected in myotubes. MyoD1 and myogenin provide precise markers for the very early identification and study of mononuclear skeletal muscle precusor cells in muscle regenerating in vivo.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Benezra R, Davis RL, Lockshon D, Turner DL, Weintraub H (1990) The protein Id: a negative regulator of helix-loop-helix DNA binding proteins. Cell 61:49–59
Bischoff R (1979) Tissue culture studies on the origin of myogenic cells during muscle regeneration in the rat. In: Mauro A (ed) Muscle regeneration. Raven Press, New York, pp 13–29
Braun T, Bober E, Buschhausen-Denker G, Kohtz S, Grzeschik KH, Arnold HH (1989) Differential expression of myogenic determination genes in muscle cells: possible autoactivation by the Myf gene products. EMBO J 8:3617–3625
Buckingham M, Lyons G, Ott M-O, Catala F (1991) Myogenesis in the mouse. J Cell Biochem 15C:2
Choi J, Costa ML, Mermelstein CS, Chagas C, Holtzer S, Holtzer H (1990) MyoD converts primary dermal fibroblasts, chondroblasts, smooth muscle, and retinal pigmented epithelial cells into striated mononucleated myoblasts and multinucleated myotubes. Proc Natl Acad Sci USA 87:7988–7992
Davis RL, Weintraub H, Lassar AB (1987) Expression of a single transfected cDNA converts fibroblasts to myoblasts. Cell 51:987–1000
Eftimie R, Brenner HR, Buonanno A (1991) Myogenin and MyoD join a family of skeletal muscle genes regulated by electrical activity. Proc Natl Acad Sci USA 88:1349–1353
Gall JG, Pardue ML (1971) Nucleic acid hybridisation in cytological preparations. Methods Enzymol 38:470–480
Grounds MD (1990) Factors controlling skeletal muscle regeneration. In: Kakulas BA, Mastaglia FL (eds) Pathogenesis and therapy of Duchenne and Becker muscular dystrophy. Raven Press, New York, pp 177–185
Ground MD (1991) Towards understanding skeletal muscle regeneration. Pathol Res Pract 187:1–22
Grounds MD, McGeachie JK (1989) A comparison of muscle precursor replication in crush injured skeletal muscle of Swiss and BALBc mice. Cell Tissue Res 255:385–391
Gunning P, Hardeman E, Wade R, Ponte W, Blau HM, Kedes L (1987) Differential patterns of transcript accumulation during human myogenesis. Mol Cell Biol 7:4100–4114
Hopwood ND, Pluck A, Gurdon JB (1989) MyoD expression in the forming somites is an early response to mesoderm induction in Xenopus embryos. EMBO J 8:3409–3417
Hughes SM, Blau HM (1990) Migration of myoblasts across basal lamina during skeletal muscle development. Nature 345:350–353
Lawrence JB, Taneja K, Singer RH (1989) Temporal resolution and sequential expression of muscle-specific genes revealed by in situ hybridisation. Dev Biol 133:235–246
Mauro A (1961) Satellite cells of skeletal muscle fibres. J Biophys Biochem Cytol 9:493–495
Mazanet R, Franzini-Armstrong C (1980) The satellite cell. In: Engel AG, Banker BQ (eds) Myology. McGraw-Hill, New York, pp 285–307
Montarras D, Pinset C, Chelly J, Kahn A, Gros F (1989) Expression of MyoD1 coincides with terminal differentiation in determined but inducible muscle cells. EMBO J 8:2203–2207
Partridge TA (1991) Myoblast transplantation: a possible therapy for inherited myopathies. Muscle Nerve 14:197–212
Partridge TA, Morgan JE, Coulton GR, Hoffman EP, Kunkel LM (1989) Conversion of mdx myofibres from dystrophin-negative to dystrophin-positive by injection of normal myoblasts. Nature 337:176–179
Phillips GD, Hoffman JR, Knighton DR (1990) Migration of myogenic cells in the rat extensor digitorum longus muscle studied with the split autograft model. Cell Tissue Res 262:81–88
Robertson TA, Papadimitriou JM, Mitchell CA, Grounds MD (1990) Fusion of myogenic cells in vivo: an ultrastructural study of regenerating murine skeletal muscle. J Struct Biol 105:170–182
Sassoon DA, Garner I, Buckingham M (1988) Transcripts of α-cardiac and α-skeletal actins are early markers for myogenesis in the mouse embryo. Development 104:155–164
Sassoon D, Lyons G, Wright WE, Lin V, Lassar A, Weintraub H, Buckingham M (1989) Expression of two myogenic regulatory factors myogenin and MyoD1 during mouse embryogensis. Nature 341:303–307
Scales JB, Olson EN, Perry M (1990) Two distinct Xenopus genes with homology to MyoD1 are expressed before somite formation in early embryogenesis. Mol Cell Biol 10:1516–1524
Schultz E, Jaryszak DL, Valliere CR (1985) Response of satellite cells to focal skeletal muscle injury. Muscle Nerve 8:217–222
Thayer MJ, Tapscott SJ, Davis RL, Wright WE, Lassar AB, Weintraub H (1989) Positive autoregulation of the myogenic determination gene MyoD1. Cell 58:241–248
Weintraub H, Davis R, Tapscott S, Thayer M, Krause M, Benezra R, Blackwell TK, Turner D, Rupp R, Hollenberg S, Shuang Y, Lassar A (1991) The MyoD gene family: nodal point during specification of the muscle cell lineage. Science 251:671–766
Wright WE, Sassoon DA, Lin VK (1989) Myogenin a factor regulating myogenesis has a domain homologous to MyoD1. Cell 56:607–617
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Grounds, M.D., Garrett, K.L., Lai, M.C. et al. Identification of skeletal muscle precursor cells in vivo by use of MyoD1 and myogenin probes. Cell Tissue Res 267, 99–104 (1992). https://doi.org/10.1007/BF00318695
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00318695