Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

Mrf4 determines skeletal muscle identity in Myf5:Myod double-mutant mice

Nature volume 431, pages 466–471 (2004)Cite this article

Abstract

In vertebrates, skeletal muscle is a model for the acquisition of cell fate from stem cells1. Two determination factors of the basic helix–loop–helix myogenic regulatory factor (MRF) family, Myf5 and Myod, are thought to direct this transition because double-mutant mice totally lack skeletal muscle fibres and myoblasts2,3,4. In the absence of these factors, progenitor cells remain multipotent and can change their fate5,6. Gene targeting studies have revealed hierarchical relationships between these and the other MRF genes, Mrf4 and myogenin, where the latter are regarded as differentiation genes7. Here we show, using an allelic series of three Myf5 mutants that differentially affect the expression of the genetically linked Mrf4 gene, that skeletal muscle is present in the new Myf5:Myod double-null mice only when Mrf4 expression is not compromised. This finding contradicts the widely held view that myogenic identity is conferred solely by Myf5 and Myod, and identifies Mrf4 as a determination gene. We revise the epistatic relationship of the MRFs, in which both Myf5 and Mrf4 act upstream of Myod to direct embryonic multipotent cells into the myogenic lineage.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Allelic series of Myf5 mutants.
Figure 2: Myod expression depends on Mrf4 in Myf5 mutants.
Figure 3: Myotome formation and desmin staining in Myf5-null and Mrf4:Myf5 double-mutant embyros.
Figure 4: Myf5:Myod double mutants make muscle.
Figure 5: Genetic hierarchy governing the acquisition of skeletal muscle identity in the mouse.

Similar content being viewed by others

References

  1. Tajbakhsh, S. Stem cells to tissue: molecular, cellular and anatomical heterogeneity in skeletal muscle. Curr. Opin. Genet. Dev. 13, 412–422 (2003)

    Article  Google Scholar 

  2. Rudnicki, M. A. et al. MyoD or myf-5 is required for the formation of skeletal muscle. Cell 75, 1351–1359 (1993)

    Article  CAS  PubMed  Google Scholar 

  3. Kaul, A., Koster, M., Neuhaus, H. & Braun, T. Myf-5 revisited: loss of early myotome formation does not lead to a rib phenotype in homozygous Myf-5 mutant mice. Cell 102, 17–19 (2000)

    Article  CAS  PubMed  Google Scholar 

  4. Kablar, B., Krastel, K., Tajbakhsh, S. & Rudnicki, M. A. Myf5 and MyoD activation define independent myogenic compartments during embryonic development. Dev. Biol. 258, 307–318 (2003)

    Article  CAS  PubMed  Google Scholar 

  5. Tajbakhsh, S., Rocancourt, D. & Buckingham, M. Muscle progenitor cells failing to respond to positional cues adopt non-myogenic fates in myf-5 null mice. Nature 384, 266–270 (1996)

    Article  ADS  CAS  PubMed  Google Scholar 

  6. Kablar, B. et al. Myogenic determination occurs independently in somites and limb buds. Dev. Biol. 206, 219–231 (1999)

    Article  CAS  PubMed  Google Scholar 

  7. Tajbakhsh, S. & Buckingham, M. The birth of muscle progenitor cells in the mouse: spatiotemporal considerations. Curr. Top. Dev. Biol. 48, 225–268 (2000)

    Article  CAS  PubMed  Google Scholar 

  8. Weintraub, H. et al. The MyoD gene family: nodal point during specification of the muscle cell lineage. Science 251, 761–766 (1991)

    Article  ADS  CAS  PubMed  Google Scholar 

  9. Braun, T., Rudnicki, M. A., Arnold, H. H. & Jaenisch, R. Targeted inactivation of the muscle regulatory gene Myf-5 results in abnormal rib development and perinatal death. Cell 71, 369–382 (1992)

    Article  CAS  PubMed  Google Scholar 

  10. Tallquist, M. D., Weismann, K. E., Hellstrom, M. & Soriano, P. Early myotome specification regulates PDGFA expression and axial skeleton development. Development 127, 5059–5070 (2000)

    CAS  PubMed  Google Scholar 

  11. Kablar, B. et al. MyoD and Myf-5 differentially regulate the development of limb versus trunk skeletal muscle. Development 124, 4729–4738 (1997)

    CAS  PubMed  Google Scholar 

  12. Summerbell, D., Halai, C. & Rigby, P. W. Expression of the myogenic regulatory factor Mrf4 precedes or is contemporaneous with that of Myf5 in the somitic bud. Mech. Dev. 117, 331–335 (2002)

    Article  CAS  PubMed  Google Scholar 

  13. Tajbakhsh, S., Rocancourt, D., Cossu, G. & Buckingham, M. Redefining the genetic hierarchies controlling skeletal myogenesis: Pax-3 and Myf-5 act upstream of MyoD. Cell 89, 127–138 (1997)

    Article  CAS  PubMed  Google Scholar 

  14. Nabeshima, Y. et al. Myogenin gene disruption results in perinatal lethality because of severe muscle defect. Nature 364, 532–535 (1993)

    Article  ADS  CAS  PubMed  Google Scholar 

  15. Rawls, A. et al. Overlapping functions of the myogenic bHLH genes MRF4 and MyoD revealed in double mutant mice. Development 125, 2349–2358 (1998)

    CAS  PubMed  Google Scholar 

  16. Moran, J. L. et al. Limbs move beyond the radical fringe. Nature 399, 742–743 (1999)

    Article  ADS  CAS  PubMed  Google Scholar 

  17. Pham, C. T., MacIvor, D. M., Hug, B. A., Heusel, J. W. & Ley, T. J. Long-range disruption of gene expression by a selectable marker cassette. Proc. Natl Acad. Sci. USA 93, 13090–13095 (1996)

    Article  ADS  CAS  PubMed  PubMed Central  Google Scholar 

  18. Olson, E. N., Arnold, H.-H., Rigby, P. W. J. & Wold, B. J. Know your neighbors: three phenotypes in null mutants of the myogenic bHLH gene MRF4. Cell 85, 1–4 (1996)

    Article  CAS  PubMed  Google Scholar 

  19. Hadchouel, J. et al. Modular long-range regulation of Myf5 reveals unexpected heterogeneity between skeletal muscles in the mouse embryo. Development 127, 4455–4467 (2000)

    CAS  PubMed  Google Scholar 

  20. Carvajal, J. J., Cox, D., Summerbell, D. & Rigby, P. W. A BAC transgenic analysis of the Mrf4/Myf5 locus reveals interdigitated elements that control activation and maintenance of gene expression during muscle development. Development 128, 1857–1868 (2001)

    CAS  PubMed  Google Scholar 

  21. Maroto, M. et al. Ectopic Pax-3 activates MyoD and Myf-5 expression in embryonic mesoderm and neural tissue. Cell 89, 139–148 (1997)

    Article  CAS  PubMed  Google Scholar 

  22. Lu, J. R. et al. Control of facial muscle development by MyoR and capsulin. Science 298, 2378–2381 (2002)

    Article  ADS  CAS  PubMed  Google Scholar 

  23. Wang, Y. & Jaenisch, R. Myogenin can substitute for Myf5 in promoting myogenesis but less efficiently. Development 124, 2507–2513 (1997)

    CAS  PubMed  Google Scholar 

  24. Bergstrom, D. A. & Tapscott, S. J. Molecular distinction between specification and differentiation in the myogenic basic helix–loop–helix transcription factor family. Mol. Cell. Biol. 21, 2404–2412 (2001)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Kress, C., Vandormael-Pournin, S., Baldacci, P., Cohen-Tannoudji, M. & Babinet, C. Nonpermissiveness for mouse embryonic stem (ES) cell derivation circumvented by a single backcross to 129/Sv strain: establishment of ES cell lines bearing the Omd conditional lethal mutation. Mamm. Genome 9, 998–1001 (1998)

    Article  CAS  PubMed  Google Scholar 

  26. Lallemand, Y., Luria, V., Haffner-Krausz, R. & Lonai, P. Maternally expressed PGK-Cre transgene as a tool for early and uniform activation of the Cre site-specific recombinase. Transgenic Res. 7, 105–112 (1998)

    Article  CAS  PubMed  Google Scholar 

  27. Tajbakhsh, S. & Buckingham, M. E. Lineage restriction of the myogenic conversion factor myf-5 in the brain. Development 121, 4077–4083 (1995)

    CAS  PubMed  Google Scholar 

  28. Bober, E. et al. The muscle regulatory gene, Myf-6, has a biphasic pattern of expression during early mouse development. J. Cell Biol. 113, 1255–1265 (1991)

    Article  CAS  PubMed  Google Scholar 

  29. Daubas, P., Tajbakhsh, S., Hadchouel, J., Primig, M. & Buckingham, M. Myf5 is a novel early axonal marker in the mouse brain and is subjected to post-transcriptional regulation in neurons. Development 127, 319–331 (2000)

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank S. Shorte, E. Perret and P. Roux at the Pasteur Dynamic Imaging Center, and C. Cimper for assistance. The laboratories of S.T. and M.B. were funded by the Pasteur Institute, CNRS, AFM, ACI Integrative Biology programme of the French Ministry. M.B. was funded by the European Union. S.T. was also funded by grants from the HFSPO, Association pour la Recherche sur le Cancer. B.G.-M. received a fellowship from the ARC, and V.S. a grant from the Fondation pour la Recherche Médicale and Pasteur Institute. L.K.-D. was funded by the HFSPO, the EU and Pasteur Institute.

Author information

Author notes

  1. Barbara Gayraud-Morel and Vasily Shinin: These authors contributed equally to the work

Authors and Affiliations

  1. Stem Cells and Development, Department of Developmental Biology, CNRS URA 2578, 25 rue du Dr Roux, 75724, Paris Cedex, 15, France

    Lina Kassar-Duchossoy, Barbara Gayraud-Morel, Danielle Gomès, Vasily Shinin & Shahragim Tajbakhsh

  2. Molecular Genetics of Development, Department of Developmental Biology, CNRS URA 2578, 25 rue du Dr Roux, 75724, Paris Cedex, 15, France

    Didier Rocancourt & Margaret Buckingham

Authors
  1. Lina Kassar-Duchossoy
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Barbara Gayraud-Morel
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Danielle Gomès
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Didier Rocancourt
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Margaret Buckingham
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Vasily Shinin
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Shahragim Tajbakhsh
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Shahragim Tajbakhsh.

Ethics declarations

Competing interests

The authors declare that they have no competing financial interests.

Supplementary information

Supplementary Data

Null status of Myf5 allelic mutants. (DOC 25 kb)

Supplementary Figure 1

Characterisation of Myf5nlacZ, Myf5GFP-P and Myf5loxP alleles. (JPG 72 kb)

Supplementary Figure 1 legend

Characterisation of Myf5nlacZ, Myf5GFP-P and Myf5loxP alleles. (DOC 21 kb)

Supplementary Figure 2

Myogenic conversion assay of Myf5 allelic constructs by transient transfections. (JPG 37 kb)

Supplementary Figure 2 legend

Myogenic conversion assay of Myf5 allelic constructs by transient transfections. (DOC 20 kb)

Supplementary Figure 3

Ex vivo analysis of Myf5:Myod mutants ; no myogenesis in embryos lacking Mrf4, Myf5 and Myod. (JPG 198 kb)

Supplementary Figure 3 legend

Ex vivo analysis of Myf5:Myod mutants ; no myogenesis in embryos lacking Mrf4, Myf5 and Myod. (DOC 21 kb)

Supplementary Methods

Explant cultures and immunofluorescence. (DOC 42 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kassar-Duchossoy, L., Gayraud-Morel, B., Gomès, D. et al. Mrf4 determines skeletal muscle identity in Myf5:Myod double-mutant mice. Nature 431, 466–471 (2004). https://doi.org/10.1038/nature02876

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature02876

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing