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Regulatory constraints in the evolution of the tetrapod limb anterior–posterior polarity

Abstract

The anterior to posterior (A–P) polarity of the tetrapod limb is determined by the confined expression of Sonic hedgehog (Shh) at the posterior margin of developing early limb buds1,2, under the control of HOX proteins encoded by gene members of both the HoxA and HoxD clusters3,4,5,6. Here, we use a set of partial deletions to show that only the last four Hox paralogy groups can elicit this response: that is, precisely those genes whose expression is excluded from most anterior limb bud cells owing to their collinear transcriptional activation. We propose that the limb A–P polarity is produced as a collateral effect of Hox gene collinearity, a process highly constrained by its crucial importance during trunk development. In this view, the co-option of the trunk collinear mechanism, along with the emergence of limbs, imposed an A–P polarity to these structures as the most parsimonious solution. This in turn further contributed to stabilize the architecture and operational mode of this genetic system.

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Figure 1: Control of Shh expression by Hoxa and Hoxd genes.
Figure 2: 5′-located Hox genes induce Shh expression in a gene-specific and dose-dependent manner.
Figure 3: Skeletons of the various mutant stocks in either the presence (top) or absence (bottom) of Shh signalling.
Figure 4: Regulatory constraints in the emergence of the limb A–P polarity.

References

  1. Saunders, J. W. & Gasselin, M. T. Ectodermal-Mesodermal Interaction in the Origin of Limb Symmetry 78–97 (Williams and Wilkins, Baltimore, 1968)

  2. Riddle, R. D., Johnson, R. L., Laufer, E. & Tabin, C. Sonic hedgehog mediates the polarizing activity of the ZPA. Cell 75, 1401–1416 (1993)

    Article  CAS  PubMed  Google Scholar 

  3. Knezevic, V. et al. Hoxd-12 differentially affects preaxial and postaxial chondrogenic branches in the limb and regulates Sonic hedgehog in a positive feedback loop. Development 124, 4523–4536 (1997)

    CAS  PubMed  Google Scholar 

  4. Charite, J., de Graaff, W., Shen, S. & Deschamps, J. Ectopic expression of Hoxb-8 causes duplication of the ZPA in the forelimb and homeotic transformation of axial structures. Cell 78, 589–601 (1994)

    Article  CAS  PubMed  Google Scholar 

  5. Zakany, J., Kmita, M. & Duboule, D. A dual role for Hox genes in limb anterior-posterior asymmetry. Science 304, 1669–1672 (2004)

    Article  ADS  CAS  PubMed  Google Scholar 

  6. Capellini, T. D. et al. Pbx1/Pbx2 requirement for distal limb patterning is mediated by the hierarchical control of Hox gene spatial distribution and Shh expression. Development 133, 2263–2273 (2006)

    Article  CAS  PubMed  Google Scholar 

  7. Laufer, E., Nelson, C. E., Johnson, R. L., Morgan, B. A. & Tabin, C. Sonic hedgehog and Fgf-4 act through a signaling cascade and feedback loop to integrate growth and patterning of the developing limb bud. Cell 79, 993–1003 (1994)

    Article  CAS  PubMed  Google Scholar 

  8. Niswander, L., Jeffrey, S., Martin, G. R. & Tickle, C. A positive feedback loop coordinates growth and patterning in the vertebrate limb. Nature 371, 609–612 (1994)

    Article  ADS  CAS  PubMed  Google Scholar 

  9. Zuniga, A., Haramis, A. P., McMahon, A. P. & Zeller, R. Signal relay by BMP antagonism controls the SHH/FGF4 feedback loop in vertebrate limb buds. Nature 401, 598–602 (1999)

    Article  ADS  CAS  PubMed  Google Scholar 

  10. Sun, X. et al. Conditional inactivation of Fgf4 reveals complexity of signalling during limb bud development. Nature Genet. 25, 83–86 (2000)

    Article  CAS  PubMed  Google Scholar 

  11. Khokha, M. K., Hsu, D., Brunet, L. J., Dionne, M. S. & Harland, R. M. Gremlin is the BMP antagonist required for maintenance of Shh and Fgf signals during limb patterning. Nature Genet. 34, 303–307 (2003)

    Article  CAS  PubMed  Google Scholar 

  12. Scherz, P. J., Harfe, B. D., McMahon, A. P. & Tabin, C. J. The limb bud Shh-Fgf feedback loop is terminated by expansion of former ZPA cells. Science 305, 396–399 (2004)

    Article  ADS  CAS  PubMed  Google Scholar 

  13. Chiang, C. et al. Manifestation of the limb prepattern: limb development in the absence of Sonic hedgehog function. Dev. Biol. 236, 421–435 (2001)

    Article  CAS  PubMed  Google Scholar 

  14. Kraus, P., Fraidenraich, D. & Loomis, C. A. Some distal limb structures develop in mice lacking Sonic hedgehog signaling. Mech. Dev. 100, 45–58 (2001)

    Article  CAS  PubMed  Google Scholar 

  15. Lewis, P. M. et al. Cholesterol modification of Sonic hedgehog is required for long-range signaling activity and effective modulation of signaling by Ptc1. Cell 105, 599–612 (2001)

    Article  CAS  PubMed  Google Scholar 

  16. Harfe, B. D. et al. Evidence for an expansion-based temporal Shh gradient in specifying vertebrate digit identities. Cell 118, 517–528 (2004)

    Article  CAS  PubMed  Google Scholar 

  17. Kmita, M. et al. Early developmental arrest of mammalian limbs lacking HoxA/HoxD gene function. Nature 435, 1113–1116 (2005)

    Article  ADS  CAS  PubMed  Google Scholar 

  18. Tarchini, B. & Duboule, D. Control of Hoxd genes' collinearity during early limb development. Dev. Cell 10, 93–103 (2006)

    Article  CAS  PubMed  Google Scholar 

  19. Shubin, N. H., Tabin, C. J. & Carroll, S. Fossils, genes and the evolution of animal limbs. Nature 388, 639–648 (1997)

    Article  ADS  CAS  PubMed  Google Scholar 

  20. Fromental-Ramain, C. et al. Specific and redundant functions of the paralogous Hoxa-9 and Hoxd-9 genes in forelimb and axial skeleton patterning. Development 122, 461–472 (1996)

    CAS  PubMed  Google Scholar 

  21. Izpisúa-Belmonte, J. C. et al. Murine genes related to the Drosophila AbdB homeotic gene are sequentially expressed during development of the posterior part of the body.. EMBO J. 10, 2279–2289 (1991)

    Article  PubMed  PubMed Central  Google Scholar 

  22. van der Hoeven, F., Zakany, J. & Duboule, D. Gene transpositions in the HoxD complex reveal a hierarchy of regulatory controls. Cell 85, 1025–1035 (1996)

    Article  CAS  PubMed  Google Scholar 

  23. Coates, M. I. The origin of vertebrate limbs. Development (Suppl.) 120, 169–180 (1994)

    Google Scholar 

  24. Kmita, M., van Der Hoeven, F., Zakany, J., Krumlauf, R. & Duboule, D. Mechanisms of Hox gene colinearity: transposition of the anterior Hoxb1 gene into the posterior HoxD complex. Genes Dev. 14, 198–211 (2000)

    CAS  PubMed  PubMed Central  Google Scholar 

  25. Tarchini, B., Huynh, T. H., Cox, G. A. & Duboule, D. HoxD cluster scanning deletions identify multiple defects leading to paralysis in the mouse mutant Ironside. Genes Dev. 19, 2862–2876 (2005)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Echelard, Y. et al. Sonic hedgehog, a member of a family of putative signaling molecules, is implicated in the regulation of CNS polarity. Cell 75, 1417–1430 (1993)

    Article  CAS  PubMed  Google Scholar 

  27. Inouye, M. Differential staining of cartilage and bone in fetal mouse skeleton by alcian blue and alizarin red. Congenital Anomalies 16, 171–173 (1976)

    Google Scholar 

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Acknowledgements

We thank N. Fraudeau and T. H. N. Huynh for technical assistance as well as C. Tabin and M. Coates for comments and suggestions. This work was supported by funds from the canton de Genève, the Louis-Jeantet foundation, the Swiss National Research Fund, the National Center for Competence in Research (NCCR) ‘Frontiers in Genetics’ and the EU programme ‘Cells into Organs’.

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Correspondence to Denis Duboule.

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Tarchini, B., Duboule, D. & Kmita, M. Regulatory constraints in the evolution of the tetrapod limb anterior–posterior polarity. Nature 443, 985–988 (2006). https://doi.org/10.1038/nature05247

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