Maintenance of muscle stem-cell quiescence by microRNA-489

Abstract

Among the key properties that distinguish adult mammalian stem cells from their more differentiated progeny is the ability of stem cells to remain in a quiescent state for prolonged periods of time1,2. However, the molecular pathways for the maintenance of stem-cell quiescence remain elusive. Here we use adult mouse muscle stem cells (satellite cells) as a model system and show that the microRNA (miRNA) pathway is essential for the maintenance of the quiescent state. Satellite cells that lack a functional miRNA pathway spontaneously exit quiescence and enter the cell cycle. We identified quiescence-specific miRNAs in the satellite-cell lineage by microarray analysis. Among these, miRNA-489 (miR-489) is highly expressed in quiescent satellite cells and is quickly downregulated during satellite-cell activation. Further analysis revealed that miR-489 functions as a regulator of satellite-cell quiescence, as it post-transcriptionally suppresses the oncogene Dek, the protein product of which localizes to the more differentiated daughter cell during asymmetric division of satellite cells and promotes the transient proliferative expansion of myogenic progenitors. Our results provide evidence of the miRNA pathway in general, and of a specific miRNA, miR-489, in actively maintaining the quiescent state of an adult stem-cell population.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Figure 1: The miRNA pathway is essential for the maintenance of satellite-cell quiescence and survival of activated satellite cells.
Figure 2: miRNA expression in purified QSCs and ASCs.
Figure 3: miRNA-489 regulates satellite-cell quiescence.
Figure 4: Targeting of Dek mRNA by miR-489 and regulation of cell-fate decision of satellite-cell progeny by Dek.

References

  1. 1

    Li, L. & Clevers, H. Coexistence of quiescent and active adult stem cells in mammals. Science 327, 542–545 (2010)

    ADS  CAS  Article  Google Scholar 

  2. 2

    Fuchs, E. The tortoise and the hair: slow-cycling cells in the stem cell race. Cell 137, 811–819 (2009)

    CAS  Article  Google Scholar 

  3. 3

    Yi, R., Poy, M. N., Stoffel, M. & Fuchs, E. A skin microRNA promotes differentiation by repressing 'stemness'. Nature 452, 225–229 (2008)

    ADS  CAS  Article  Google Scholar 

  4. 4

    Tiscornia, G. & Izpisua Belmonte, J. C. MicroRNAs in embryonic stem cell function and fate. Genes Dev. 24, 2732–2741 (2010)

    CAS  Article  Google Scholar 

  5. 5

    Nishijo, K. et al. Biomarker system for studying muscle, stem cells, and cancer in vivo. FASEB J. 23, 2681–2690 (2009)

    CAS  Article  Google Scholar 

  6. 6

    Harfe, B. D., McManus, M. T., Mansfield, J. H., Hornstein, E. & Tabin, C. J. The RNaseIII enzyme Dicer is required for morphogenesis but not patterning of the vertebrate limb. Proc. Natl Acad. Sci. USA 102, 10898–10903 (2005)

    ADS  CAS  Article  Google Scholar 

  7. 7

    Srinivas, S. et al. Cre reporter strains produced by targeted insertion of EYFP and ECFP into the ROSA26 locus. BMC Dev. Biol. 1, 4 (2001).f

    CAS  Article  Google Scholar 

  8. 8

    Morgan, J. E. & Partridge, T. A. Muscle satellite cells. Int. J. Biochem. Cell Biol. 35, 1151–1156 (2003)

    CAS  Article  Google Scholar 

  9. 9

    Friedman, R. C., Farh, K. K., Burge, C. B. & Bartel, D. P. Most mammalian mRNAs are conserved targets of microRNAs. Genome Res. 19, 92–105 (2009)

    CAS  Article  Google Scholar 

  10. 10

    Fukada, S. et al. Molecular signature of quiescent satellite cells in adult skeletal muscle. Stem Cells 25, 2448–2459 (2007)

    CAS  Article  Google Scholar 

  11. 11

    van Rooij, E. et al. Control of stress-dependent cardiac growth and gene expression by a microRNA. Science 316, 575–579 (2007)

    ADS  CAS  Article  Google Scholar 

  12. 12

    Olguin, H. C. & Olwin, B. B. Pax-7 up-regulation inhibits myogenesis and cell cycle progression in satellite cells: a potential mechanism for self-renewal. Dev. Biol. 275, 375–388 (2004)

    CAS  Article  Google Scholar 

  13. 13

    Tanaka, K. K. et al. Syndecan-4-expressing muscle progenitor cells in the SP engraft as satellite cells during muscle regeneration. Cell Stem Cell 4, 217–225 (2009)

    CAS  Article  Google Scholar 

  14. 14

    Zammit, P. S., Partridge, T. A. & Yablonka-Reuveni, Z. The skeletal muscle satellite cell: the stem cell that came in from the cold. J. Histochem. Cytochem. 54, 1177–1191 (2006)

    CAS  Article  Google Scholar 

  15. 15

    Krutzfeldt, J. et al. Silencing of microRNAs in vivo with 'antagomirs'. Nature 438, 685–689 (2005)

    ADS  Article  Google Scholar 

  16. 16

    Grimson, A. et al. MicroRNA targeting specificity in mammals: determinants beyond seed pairing. Mol. Cell 27, 91–105 (2007)

    CAS  Article  Google Scholar 

  17. 17

    Khodadoust, M. S. et al. Melanoma proliferation and chemoresistance controlled by the DEK oncogene. Cancer Res. 69, 6405–6413 (2009)

    CAS  Article  Google Scholar 

  18. 18

    Soares, L. M., Zanier, K., Mackereth, C., Sattler, M. & Valcarcel, J. Intron removal requires proofreading of U2AF/3′ splice site recognition by DEK. Science 312, 1961–1965 (2006)

    ADS  Article  Google Scholar 

  19. 19

    Zammit, P. S. et al. Muscle satellite cells adopt divergent fates: a mechanism for self-renewal? J. Cell Biol. 166, 347–357 (2004)

    CAS  Article  Google Scholar 

  20. 20

    Zammit, P. S. et al. Pax7 and myogenic progression in skeletal muscle satellite cells. J. Cell Sci. 119, 1824–1832 (2006)

    CAS  Article  Google Scholar 

  21. 21

    Conboy, M. J., Karasov, A. O. & Rando, T. A. High incidence of non-random template strand segregation and asymmetric fate determination in dividing stem cells and their progeny. PLoS Biol. 5, e102 (2007)

    Article  Google Scholar 

  22. 22

    Shinin, V., Gayraud-Morel, B., Gomes, D. & Tajbakhsh, S. Asymmetric division and cosegregation of template DNA strands in adult muscle satellite cells. Nature Cell Biol. 8, 677–687 (2006)

    CAS  Article  Google Scholar 

  23. 23

    Rosenblatt, J. D., Lunt, A. I., Parry, D. J. & Partridge, T. A. Culturing satellite cells from living single muscle fibre explants. In vitro Cell Dev. Biol. Anim. 31, 773–779 (1995)

    CAS  Article  Google Scholar 

  24. 24

    Bertoni, C. et al. Enhancement of plasmid-mediated gene therapy for muscular dystrophy by directed plasmid integration. Proc. Natl Acad. Sci. USA 103, 419–424 (2006)

    ADS  CAS  Article  Google Scholar 

  25. 25

    Pfaffl, M. W. A new mathematical model for relative quantification in real-time RT–PCR. Nucleic Acids Res. 29, e45 (2001)

    CAS  Article  Google Scholar 

Download references

Acknowledgements

We thank the members of the Rando laboratory for comments and discussions. We thank B. Olwin for providing the syndecan 4 antibody. This work was supported by the Glenn Foundation for Medical Research and by grants from the National Institutes of Health (NIH) (P01 AG036695, R01 AG23806 (R37 MERIT Award), R01 AR062185 and DP1 OD000392 (an NIH Director's Pioneer Award)) and the Department of Veterans Affairs (Merit Review) to T.A.R.

Author information

Affiliations

Authors

Contributions

T.H.C. and T.A.R. conceived the study. T.H.C., N.L.Q., G.W.C., L.L. and T.A.R. designed the experiments. T.H.C., B.Y. and L.L. performed all FACS analyses. T.H.C., N.L.Q., G.W.C., L.P., A.E., B.Y. and P.H. performed the experiments and analysed the experimental data. T.H.C. and T.A.R. wrote the manuscript.

Corresponding author

Correspondence to Thomas A. Rando.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-14 with legends and Supplementary Table 1. (PDF 2023 kb)

PowerPoint slides

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Cheung, T., Quach, N., Charville, G. et al. Maintenance of muscle stem-cell quiescence by microRNA-489. Nature 482, 524–528 (2012). https://doi.org/10.1038/nature10834

Download citation

Further reading

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

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