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The microRNA miR-196 acts upstream of Hoxb8 and Shh in limb development

Nature volume 438, pages 671–674 (2005)Cite this article

Abstract

MicroRNAs (miRNAs) are an abundant class of gene regulatory molecules (reviewed in refs 1, 2). Although computational work indicates that miRNAs repress more than a third of human genes3, their roles in vertebrate development are only now beginning to be determined. Here we show that miR-196 acts upstream of Hoxb8 and Sonic hedgehog (Shh) in vivo in the context of limb development, thereby identifying a previously observed but uncharacterized inhibitory activity that operates specifically in the hindlimb. Our data indicate that miR-196 functions in a fail-safe mechanism to assure the fidelity of expression domains that are primarily regulated at the transcriptional level, supporting the idea that many vertebrate miRNAs may function as a secondary level of gene regulation.

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Figure 1: Activity downstream of Dicer inhibits RA-induced expression of Hoxb8 in mouse hindlimbs.
Figure 2: Hindlimb-specific expression of miR-196.
Figure 3: miR-196 downregulates Hoxb8 accumulation.
Figure 4: miR-196 downregulates Shh in the chick forelimb.

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Acknowledgements

We thank E. McGlinn and S. Nissim for critically reading the manuscript. This work was funded by grants from the NIH (to C.J.T. and to D.P.B.). E.H. was supported by a ‘Dorot’ fellowship and ‘Bikura’ award. J.H.M. is supported by a Kirchstein postdoctoral fellowship.

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Authors and Affiliations

  1. Department of Genetics, Harvard Medical School, 77 Avenue Louis Pasteur, Massachusetts, 02115, Boston, USA

    Eran Hornstein, Jennifer H. Mansfield, Jimmy Kuang-Hsien Hu & Clifford J. Tabin

  2. Whitehead Institute for Biomedical Research, Massachusetts Institute of Technology, 9 Cambridge Center, Massachusetts, 02142, Cambridge, USA

    Soraya Yekta, Scott Baskerville & David P. Bartel

  3. Department of Molecular Genetics and Microbiology, University of Florida College of Medicine, Florida, 32610, Gainesville, USA

    Brian D. Harfe

  4. Department of Microbiology and Immunology, Diabetes Center, University of California at San Francisco, California, 94143, San Francisco, USA

    Michael T. McManus

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  1. Eran Hornstein
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  9. Clifford J. Tabin
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Corresponding author

Correspondence to Clifford J. Tabin.

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Supplementary information

Supplementary Table 1

miRNA that are differentially expressed in limb buds. List of miRNAs that came up as differentially expressed in mouse E10.5 limb buds. Direct analysis of hind limb vs. forelimb probes (see methods) yielded four miRNAs that are apparently differentially expressed twofold or more in the hindlimb. Eight other miRNAs appear to be more abundant in forelimb bud. (RTF 27 kb)

Supplementary Figure 1

Limb-specific Cre recombinase and Dicer knockout limbs. A transgenic mouse expressing Cre recombinase driven by a Prx-1 enhancer. (a) A scheme for excision of a critical element in the Dicer conditional allele and Expression of genes that are not predicted to be downstream of Dicer (JPG 150 kb)

Supplementary Figure 2

Infection of RCAS::miR-196 in the chick limb bud did not affect several mesenchymal genes outside the Hoxb-8 pathway. (JPG 102 kb)

Supplementary Figure 3

Expression pattern of chick Hoxa-7 mRNA in wild-type embryos. (JPG 105 kb)

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Hornstein, E., Mansfield, J., Yekta, S. et al. The microRNA miR-196 acts upstream of Hoxb8 and Shh in limb development. Nature 438, 671–674 (2005). https://doi.org/10.1038/nature04138

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