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.

Zebrafish miR-214 modulates Hedgehog signaling to specify muscle cell fate

Nature Genetics volume 39, pages 259–263 (2007)Cite this article

Abstract

Numerous microRNAs (miRNAs) have been discovered in the genomes of higher eukaryotes, and functional studies indicate that they are important during development. However, little is known concerning the function of individual miRNAs. We approached this problem in zebrafish by combining identification of miRNA expression, functional analyses and experimental validation of potential targets. We show that miR-214 is expressed during early segmentation stages in somites and that varying its expression alters the expression of genes regulated by Hedgehog signaling. Inhibition of miR-214 results in a reduction or loss of slow-muscle cell types. We show that su(fu) mRNA, encoding a negative regulator of Hedgehog signaling, is targeted by miR-214. Through regulation of su(fu), miR-214 enables precise specification of muscle cell types by sharpening cellular responses to Hedgehog.

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

Relevant articles

Open Access articles citing this article.

Access options

Rent or buy this article

Get just this article for as long as you need it

$39.95

Learn more

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

Figure 1: miR-214 participates in somitogenesis.
Figure 2: Modulation of miR-214 expression alters Hedgehog-mediated cell fate specification.
Figure 3: su(fu) is a target of miR-214.
Figure 4: Rescue of the miR-214MO phenotype by simultaneous inhibition of su(fu) expression.
Figure 5: Modulation of Hedgehog (Hh) signaling in somite cells by miR-214.

References

  1. Giraldez, A.J. et al. MicroRNAs regulate brain morphogenesis in zebrafish. Science 308, 833–838 (2005).

    Article  CAS  PubMed  Google Scholar 

  2. Wienholds, E. et al. MicroRNA expression in zebrafish embryonic development. Science 309, 310–311 (2005).

    Article  CAS  PubMed  Google Scholar 

  3. Nasevicius, A. & Ekker, S.C. Effective targeted gene 'knockdown' in zebrafish. Nat. Genet. 26, 216–220 (2000).

    Article  CAS  PubMed  Google Scholar 

  4. Lewis, K.E. et al. Control of muscle cell-type specification in the zebrafish embryo by hedgehog signalling. Dev. Biol. 216, 469–480 (1999).

    Article  CAS  PubMed  Google Scholar 

  5. Ingham, P.W. & Kim, H.R. Hedgehog signalling and the specification of muscle cell identity in the Zebrafish embryo. Exp. Cell Res. 306, 336–342 (2005).

    Article  CAS  PubMed  Google Scholar 

  6. Wolff, C., Roy, S. & Ingham, P.W. Multiple muscle cell identities induced by distinct levels and timing of hedgehog activity in the zebrafish embryo. Curr. Biol. 13, 1169–1181 (2003).

    Article  CAS  PubMed  Google Scholar 

  7. Feng, X., Adiarte, E.G. & Devoto, S.H. Hedgehog acts directly on the zebrafish dermomyotome to promote myogenic differentiation. Dev. Biol. 300, 736–746 (2006).

    Article  CAS  PubMed  Google Scholar 

  8. Henry, C.A. & Amacher, S.L. Zebrafish slow muscle cell migration induces a wave of fast muscle morphogenesis. Dev. Cell 7, 917–923 (2004).

    Article  CAS  PubMed  Google Scholar 

  9. Goodrich, L.V., Johnson, R.L., Milenkovic, L., McMahon, J.A. & Scott, M.P. Conservation of the hedgehog/patched signaling pathway from flies to mice: induction of a mouse patched gene by Hedgehog. Genes Dev. 10, 301–312 (1996).

    Article  CAS  PubMed  Google Scholar 

  10. Roy, S., Wolff, C. & Ingham, P.W. The u-boot mutation identifies a Hedgehog-regulated myogenic switch for fiber-type diversification in the zebrafish embryo. Genes Dev. 15, 1563–1576 (2001).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Schauerte, H.E. et al. Sonic hedgehog is not required for the induction of medial floor plate cells in the zebrafish. Development 125, 2983–2993 (1998).

    CAS  PubMed  Google Scholar 

  12. Schafer, M. et al. Hedgehog and retinoid signalling confines nkx2.2b expression to the lateral floor plate of the zebrafish trunk. Mech. Dev. 122, 43–56 (2005).

    Article  PubMed  Google Scholar 

  13. Park, H.-C., Shin, J. & Appel, B. Spatial and temporal regulation of ventral spinal cord precursor specification by Hedgehog signaling. Development 131, 5959–5969 (2004).

    Article  CAS  PubMed  Google Scholar 

  14. Chen, P.Y. et al. The developmental miRNA profiles of zebrafish as determined by small RNA cloning. Genes Dev. 19, 1288–1293 (2005).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Koudijs, M.J. et al. The Zebrafish mutants dre, uki, and lep encode negative regulators of the Hedgehog signaling pathway. PLoS Genet. 1, e19 (2005).

    Article  PubMed  PubMed Central  Google Scholar 

  16. Dunaeva, M., Michelson, P., Kogerman, P. & Toftgard, R. Characterization of the physical interaction of Gli proteins with SUFU Proteins. J. Biol. Chem. 278, 5116–5122 (2003).

    Article  CAS  PubMed  Google Scholar 

  17. Bartel, D.P. & Chen, C.Z. Micromanagers of gene expression: the potentially widespread influence of metazoan microRNAs. Nat. Rev. Genet. 5, 396–400 (2004).

    Article  CAS  PubMed  Google Scholar 

  18. Rupp, R.A., Snider, L. & Weintraub, H. Xenopus embryos regulate the nuclear localization of XMyoD. Genes Dev. 8, 1311–1323 (1994).

    Article  CAS  PubMed  Google Scholar 

  19. Sempere, L.F., Sokol, N.S., Dubrovsky, E.B., Berger, E.M. & Ambros, V. Temporal regulation of microRNA expression in Drosophila melanogaster mediated by hormonal signals and broad-Complex gene activity. Dev. Biol. 259, 9–18 (2003).

    Article  CAS  PubMed  Google Scholar 

  20. Thisse, C., Thisse, B., Schilling, T.F. & Postlethwait, J.H. Structure of the zebrafish snail1 gene and its expression in wild-type, spadetail and no tail mutant embryos. Development 119, 1203–1215 (1993).

    CAS  PubMed  Google Scholar 

  21. Barnfield, P.C., Zhang, X., Thanabalasingham, V., Yoshida, M. & Hui, C.-c. Negative regulation of Gli1 and Gli2 activator function by Suppressor of fused through multiple mechanisms. Differentiation 73, 397–405 (2005).

    Article  CAS  PubMed  Google Scholar 

  22. Topczewska, J.M. et al. The winged helix transcription factor Foxc1a is essential for somitogenesis in zebrafish. Genes Dev. 15, 2483–2493 (2001).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

The authors would like to thank Y. Thu, C. Yin and E. Tillman for experimental assistance, reagents and technical advice. Fluorescent confocal microscopy was possible through use of Vanderbilt Cell-Imaging Shared Resource equipment. Mouse monoclonal antibody F59 against slow myosin HC was a gift from F. Stockdale, Stanford University), and rabbit polyclonal antibodies against chick Engrailed were a gift from A. Joyner (Skirball Institute, New York University). Requests for materials should be addressed to J.G.P. (james.g.patton@vanderbilt.edu). This work was supported by grants from the Vanderbilt Zebrafish Initiative and the NIH (GM 075790). A.F. was supported in part by T32 GM 008554.

Author information

Authors and Affiliations

  1. Department of Biological Sciences, Vanderbilt University, Nashville, 37235, Tennessee, USA

    Alex S Flynt, Nan Li, Elizabeth J Thatcher, Lilianna Solnica-Krezel & James G Patton

Authors
  1. Alex S Flynt
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Nan Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Elizabeth J Thatcher
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Lilianna Solnica-Krezel
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. James G Patton
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

A.S.F., L.S.-K. and J.G.P. conceived and designed all experiments and wrote the paper. A.S.F. performed all experiments with help from N.L. on Figure 3 and help from E.J.T. with the developmental miRNA microarrays and supplementary statistics.

Corresponding author

Correspondence to James G Patton.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Morpholino-mediated inhibition of miR-214. (PDF 217 kb)

Supplementary Fig. 2

miR-214 regulates hedgehog signaling. (PDF 722 kb)

Supplementary Table 1

Sequences of oligonucleotides. (PDF 49 kb)

Supplementary Table 2

Summary of 95% c.i., kurtosis and skewness scores for values listed in Table 1. (PDF 56 kb)

Supplementary Note (PDF 63 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Flynt, A., Li, N., Thatcher, E. et al. Zebrafish miR-214 modulates Hedgehog signaling to specify muscle cell fate. Nat Genet 39, 259–263 (2007). https://doi.org/10.1038/ng1953

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ng1953

This article is cited by

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