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


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.

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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.


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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. ( 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.

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Authors and Affiliations



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.

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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)

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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).

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