Letter | Published:

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

Nature Genetics volume 39, pages 259263 (2007) | Download Citation

Subjects

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.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

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

  2. 2.

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

  3. 3.

    & Effective targeted gene 'knockdown' in zebrafish. Nat. Genet. 26, 216–220 (2000).

  4. 4.

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

  5. 5.

    & Hedgehog signalling and the specification of muscle cell identity in the Zebrafish embryo. Exp. Cell Res. 306, 336–342 (2005).

  6. 6.

    , & Multiple muscle cell identities induced by distinct levels and timing of hedgehog activity in the zebrafish embryo. Curr. Biol. 13, 1169–1181 (2003).

  7. 7.

    , & Hedgehog acts directly on the zebrafish dermomyotome to promote myogenic differentiation. Dev. Biol. 300, 736–746 (2006).

  8. 8.

    & Zebrafish slow muscle cell migration induces a wave of fast muscle morphogenesis. Dev. Cell 7, 917–923 (2004).

  9. 9.

    , , , & 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).

  10. 10.

    , & The u-boot mutation identifies a Hedgehog-regulated myogenic switch for fiber-type diversification in the zebrafish embryo. Genes Dev. 15, 1563–1576 (2001).

  11. 11.

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

  12. 12.

    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).

  13. 13.

    , & Spatial and temporal regulation of ventral spinal cord precursor specification by Hedgehog signaling. Development 131, 5959–5969 (2004).

  14. 14.

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

  15. 15.

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

  16. 16.

    , , & Characterization of the physical interaction of Gli proteins with SUFU Proteins. J. Biol. Chem. 278, 5116–5122 (2003).

  17. 17.

    & Micromanagers of gene expression: the potentially widespread influence of metazoan microRNAs. Nat. Rev. Genet. 5, 396–400 (2004).

  18. 18.

    , & Xenopus embryos regulate the nuclear localization of XMyoD. Genes Dev. 8, 1311–1323 (1994).

  19. 19.

    , , , & Temporal regulation of microRNA expression in Drosophila melanogaster mediated by hormonal signals and broad-Complex gene activity. Dev. Biol. 259, 9–18 (2003).

  20. 20.

    , , & Structure of the zebrafish snail1 gene and its expression in wild-type, spadetail and no tail mutant embryos. Development 119, 1203–1215 (1993).

  21. 21.

    , , , & Negative regulation of Gli1 and Gli2 activator function by Suppressor of fused through multiple mechanisms. Differentiation 73, 397–405 (2005).

  22. 22.

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

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

Affiliations

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

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

Authors

  1. Search for Alex S Flynt in:

  2. Search for Nan Li in:

  3. Search for Elizabeth J Thatcher in:

  4. Search for Lilianna Solnica-Krezel in:

  5. Search for James G Patton in:

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.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to James G Patton.

Supplementary information

PDF files

  1. 1.

    Supplementary Fig. 1

    Morpholino-mediated inhibition of miR-214.

  2. 2.

    Supplementary Fig. 2

    miR-214 regulates hedgehog signaling.

  3. 3.

    Supplementary Table 1

    Sequences of oligonucleotides.

  4. 4.

    Supplementary Table 2

    Summary of 95% c.i., kurtosis and skewness scores for values listed in Table 1.

  5. 5.

    Supplementary Note

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/ng1953

Further reading