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.

  • Article
  • Published:

Cooperative and antagonistic interactions between Sall4 and Tbx5 pattern the mouse limb and heart

Nature Genetics volume 38, pages 175–183 (2006)Cite this article

Abstract

Human mutations in TBX5, a gene encoding a T-box transcription factor, and SALL4, a gene encoding a zinc-finger transcription factor, cause similar upper limb and heart defects. Here we show that Tbx5 regulates Sall4 expression in the developing mouse forelimb and heart; mice heterozygous for a gene trap allele of Sall4 show limb and heart defects that model human disease. Tbx5 and Sall4 interact both positively and negatively to finely regulate patterning and morphogenesis of the anterior forelimb and heart. Thus, a positive and negative feed-forward circuit between Tbx5 and Sall4 ensures precise patterning of embryonic limb and heart and provides a unifying mechanism for heart/hand syndromes.

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

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

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

Figure 1: Sall4 and Tbx5 expression in mouse development.
Figure 2: Tbx5 regulates Sall4 expression in forelimb and in heart.
Figure 3: Limb defects in Sall4GT/+ mice.
Figure 4: Genetic interaction between Tbx5 and Sall4 in patterning the forelimb.
Figure 5: Sall4 and Tbx5 regulate heart patterning.
Figure 6: Altered gene expression in mutant hearts.
Figure 7: Sall4 and Tbx5 interact to synergistically activate the Fgf10 promoter.
Figure 8: Cooperative and counteracting interactions between Sall4 and Tbx5 on cardiac promoters.

Similar content being viewed by others

Accession codes

Accessions

GenBank/EMBL/DDBJ

References

  1. Seidman, J.G. & Seidman, C. Transcription factor haploinsufficiency: when half a loaf is not enough. J. Clin. Invest. 109, 451–455 (2002).

    Article  CAS  Google Scholar 

  2. Bruneau, B.G. Transcriptional regulation of vertebrate cardiac morphogenesis. Circ. Res. 90, 509–519 (2002).

    Article  Google Scholar 

  3. Garg, V. et al. GATA4 mutations cause human congenital heart defects and reveal an interaction with TBX5. Nature 424, 443–447 (2003).

    Article  CAS  Google Scholar 

  4. Bruneau, B.G. et al. A murine model of Holt-Oram syndrome defines roles of the T-box transcription factor Tbx5 in cardiogenesis and disease. Cell 106, 709–721 (2001).

    Article  CAS  Google Scholar 

  5. Hiroi, Y. et al. Tbx5 associates with Nkx2–5 and synergistically promotes cardiomyocyte differentiation. Nat. Genet. 28, 276–280 (2001).

    Article  CAS  Google Scholar 

  6. Mori, A.D. & Bruneau, B.G. TBX5 mutations and congenital heart disease: Holt-Oram syndrome revealed. Curr. Opin. Cardiol. 19, 211–215 (2004).

    Article  Google Scholar 

  7. Poznanski, A.K., Gall, J.C. Jr . & Stern, A.M. Skeletal manifestations of the Holt-Oram syndrome. Radiology 94, 45–53 (1970).

    Article  CAS  Google Scholar 

  8. Basson, C.T. et al. The clinical and genetic spectrum of the Holt-Oram syndrome (heart-hand syndrome). N. Engl. J. Med. 330, 885–891 (1994).

    Article  CAS  Google Scholar 

  9. Newbury-Ecob, R.A., Leanage, R., Raeburn, J.A. & Young, I.D. Holt-Oram syndrome: a clinical genetic study. J. Med. Genet. 33, 300–307 (1996).

    Article  CAS  Google Scholar 

  10. Tickle, C. Patterning systems–from one end of the limb to the other. Dev. Cell 4, 449–458 (2003).

    Article  CAS  Google Scholar 

  11. Litingtung, Y., Dahn, R.D., Li, Y., Fallon, J.F. & Chiang, C. Shh and Gli3 are dispensable for limb skeleton formation but regulate digit number and identity. Nature 418, 979–983 (2002).

    Article  CAS  Google Scholar 

  12. te Welscher, P. et al. Progression of vertebrate limb development through SHH-mediated counteraction of GLI3. Science 298, 827–830 (2002).

    Article  CAS  Google Scholar 

  13. Drossopoulou, G. et al. A model for anteroposterior patterning of the vertebrate limb based on sequential long- and short-range Shh signalling and Bmp signalling. Development 127, 1337–1348 (2000).

    CAS  PubMed  Google Scholar 

  14. Dahn, R.D. & Fallon, J.F. Interdigital regulation of digit identity and homeotic transformation by modulated BMP signaling. Science 289, 438–441 (2000).

    Article  CAS  Google Scholar 

  15. Suzuki, T., Takeuchi, J., Koshiba-Takeuchi, K. & Ogura, T. Tbx genes specify posterior digit identity through Shh and BMP signaling. Dev. Cell 6, 43–53 (2004).

    Article  CAS  Google Scholar 

  16. 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  Google Scholar 

  17. Ahn, S. & Joyner, A.L. Dynamic changes in the response of cells to positive hedgehog signaling during mouse limb patterning. Cell 118, 505–516 (2004).

    Article  CAS  Google Scholar 

  18. 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  Google Scholar 

  19. Bamshad, M. et al. Mutations in human TBX3 alter limb, apocrine and genital development in ulnar-mammary syndrome. Nat. Genet. 16, 311–315 (1997).

    Article  CAS  Google Scholar 

  20. Davenport, T.G., Jerome-Majewska, L.A. & Papaioannou, V.E. Mammary gland, limb and yolk sac defects in mice lacking Tbx3, the gene mutated in human ulnar mammary syndrome. Development 130, 2263–2273 (2003).

    Article  CAS  Google Scholar 

  21. Kohlhase, J. et al. Okihiro syndrome is caused by SALL4 mutations. Hum. Mol. Genet. 11, 2979–2987 (2002).

    Article  CAS  Google Scholar 

  22. Al-Baradie, R. et al. Duane Radial Ray syndrome (Okihiro syndrome) maps to 20q13 and results from mutations in SALL4, a new member of the SAL family. Am. J. Hum. Genet. 71, 1195–1199 (2002).

    Article  CAS  Google Scholar 

  23. Kohlhase, J. et al. Mutations at the SALL4 locus on chromosome 20 result in a range of clinically overlapping phenotypes, including Okihiro syndrome, Holt-Oram syndrome, acro-renal-ocular syndrome, and patients previously reported to represent thalidomide embryopathy. J. Med. Genet. 40, 473–478 (2003).

    Article  CAS  Google Scholar 

  24. Borozdin, W. et al. Novel mutations in the gene SALL4 provide further evidence for acro-renal-ocular and Okihiro syndromes being allelic entities, and extend the phenotypic spectrum. J. Med. Genet. 41, e102 (2004).

    Article  CAS  Google Scholar 

  25. Brassington, A.M. et al. Expressivity of Holt-Oram syndrome is not predicted by TBX5 genotype. Am. J. Hum. Genet. 73, 74–85 (2003).

    Article  CAS  Google Scholar 

  26. Kohlhase, J. et al. Cloning and expression analysis of SALL4, the murine homologue of the gene mutated in Okihiro syndrome. Cytogenet. Genome Res. 98, 274–277 (2002).

    Article  CAS  Google Scholar 

  27. Spitz, F., Gonzalez, F. & Duboule, D. A global control region defines a chromosomal regulatory landscape containing the HoxD cluster. Cell 113, 405–417 (2003).

    Article  CAS  Google Scholar 

  28. Pizard, A. et al. Connexin 40, a target of transcription factor Tbx5, patterns wrist, digits and sternum. Mol. Cell. Biol. 25, 5073–5083 (2005).

    Article  CAS  Google Scholar 

  29. Sekine, K. et al. Fgf10 is essential for limb and lung formation. Nat. Genet. 21, 138–141 (1999).

    Article  CAS  Google Scholar 

  30. Naiche, L.A. & Papaioannou, V.E. Loss of Tbx4 blocks hindlimb development and affects vascularization and fusion of the allantois. Development 130, 2681–2693 (2003).

    Article  CAS  Google Scholar 

  31. Szeto, D.P. et al. Role of the Bicoid-related homeodomain factor Pitx1 in specifying hindlimb morphogenesis and pituitary development. Genes Dev. 13, 484–494 (1999).

    Article  CAS  Google Scholar 

  32. Logan, M. & Tabin, C.J. Role of Pitx1 upstream of Tbx4 in specification of hindlimb identity. Science 283, 1736–1739 (1999).

    Article  CAS  Google Scholar 

  33. Lanctot, C., Moreau, A., Chamberland, M., Tremblay, M.L. & Drouin, J. Hindlimb patterning and mandible development require the Ptx1 gene. Development 126, 1805–1810 (1999).

    CAS  PubMed  Google Scholar 

  34. Agarwal, P. et al. Tbx5 is essential for forelimb bud initiation following patterning of the limb field in the mouse embryo. Development 130, 623–633 (2003).

    Article  CAS  Google Scholar 

  35. Gibson-Brown, J.J. et al. Evidence of a role for T-box genes in the evolution of limb morphogenesis and the specification of forelimb/hindlimb identity. Mech. Dev. 56, 93–101 (1996).

    Article  CAS  Google Scholar 

  36. Logan, M. Finger or toe: the molecular basis of limb identity. Development 130, 6401–6410 (2003).

    Article  CAS  Google Scholar 

  37. Habets, P.E. et al. Cooperative action of Tbx2 and Nkx2.5 inhibits ANF expression in the atrioventricular canal: implications for cardiac chamber formation. Genes Dev. 16, 1234–1246 (2002).

    Article  CAS  Google Scholar 

  38. Hoogaars, W.M. et al. The transcriptional repressor Tbx3 delineates the developing central conduction system of the heart. Cardiovasc. Res. 62, 489–499 (2004).

    Article  CAS  Google Scholar 

  39. Ghosh, T.K. et al. Characterization of the TBX5 binding site and analysis of mutations that cause Holt-Oram syndrome. Hum. Mol. Genet. 10, 1983–1994 (2001).

    Article  CAS  Google Scholar 

  40. Fan, C., Liu, M. & Wang, Q. Functional analysis of TBX5 missense mutations associated with Holt-Oram syndrome. J. Biol. Chem. 278, 8780–8785 (2003).

    Article  CAS  Google Scholar 

  41. Durocher, D., Charron, F., Warren, R., Schwartz, R.J. & Nemer, M. The cardiac transcription factors Nkx2–5 and GATA-4 are mutual cofactors. EMBO J. 16, 5687–5696 (1997).

    Article  CAS  Google Scholar 

  42. Minguillon, C., Del Buono, J. & Logan, M.P. Tbx5 and Tbx4 are not sufficient to determine limb-specific morphologies but have common roles in initiating limb outgrowth. Dev. Cell 8, 75–84 (2005).

    Article  CAS  Google Scholar 

  43. Rallis, C. et al. Tbx5 is required for forelimb bud formation and continued outgrowth. Development 130, 2741–2751 (2003).

    Article  CAS  Google Scholar 

  44. Shen-Orr, S.S., Milo, R., Mangan, S. & Alon, U. Network motifs in the transcriptional regulation network of Escherichia coli. Nat. Genet. 31, 64–68 (2002).

    Article  CAS  Google Scholar 

  45. Mangan, S. & Alon, U. Structure and function of the feed-forward loop network motif. Proc. Natl. Acad. Sci. USA 100, 11980–11985 (2003).

    Article  CAS  Google Scholar 

  46. Boyer, L.A. et al. Core transcriptional regulatory circuitry in human embryonic stem cells. Cell 122, 947–956 (2005).

    Article  CAS  Google Scholar 

  47. Kaufman, M.H. The Atlas of Mouse Development (Academic, London, 1992).

    Google Scholar 

  48. Lickert, H. et al. Baf60c is essential for function of BAF chromatin remodelling complexes in heart development. Nature 432, 107–112 (2004).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We are grateful to A. Mori for help with statistics and quantitative RT-PCR. We also thank V. Christoffels, M. Nemer and T. Ogura for expression vectors and reporter constructs, and A. Brown, D. Duboule, M. Logan, G. Martin and C. Oka for in situ probes. This work was supported by grants from the Canadian Institutes of Health Research (B.G.B., C.c.H), the Heart and Stroke Foundation of Ontario (B.G.B.) and the March of Dimes Birth Defects Foundation (B.G.B.). K.K.-T. was supported by the Uehara Memorial Foundation. J.K.T holds a long-term fellowship from the Human Frontiers Science Program and was partly supported by the Mochida Memorial Foundation for Medical and Pharmaceutical Research. B.G.B. holds a Canada Research Chair in Developmental Cardiology.

Author information

Author notes

  1. Deepak Srivastava

    Present address: Gladstone Institute of Cardiovascular Disease, University of California, San Francisco, California, 94158, USA

  2. Kazuko Koshiba-Takeuchi and Jun K Takeuchi: These authors contributed equally to this work.

Authors and Affiliations

  1. Programs in Cardiovascular Research, The Hospital for Sick Children, Toronto, M5G 1X8, Ontario, Canada

    Kazuko Koshiba-Takeuchi, Jun K Takeuchi, Eric P Arruda & Benoit G Bruneau

  2. Developmental Biology, The Hospital for Sick Children, Toronto, M5G 1X8, Ontario, Canada

    Kazuko Koshiba-Takeuchi, Jun K Takeuchi, Eric P Arruda, Rong Mo, Chi-chung Hui & Benoit G Bruneau

  3. The Heart and Stroke/Richard Lewar Centre of Excellence, University of Toronto, Toronto, M5S 1A8, Ontario, Canada

    Kazuko Koshiba-Takeuchi, Jun K Takeuchi, Eric P Arruda & Benoit G Bruneau

  4. Department of Molecular and Medical Genetics, University of Toronto, Toronto, M5S 1A8, Ontario, Canada

    Eric P Arruda & Chi-chung Hui

  5. Departments of Pediatrics and Molecular Biology, University of Texas Southwestern Medical Center at Dallas, Dallas, 75390-9148, Texas, USA

    Irfan S Kathiriya & Deepak Srivastava

Authors
  1. Kazuko Koshiba-Takeuchi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jun K Takeuchi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Eric P Arruda
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Irfan S Kathiriya
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Rong Mo
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Chi-chung Hui
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Deepak Srivastava
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Benoit G Bruneau
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Benoit G Bruneau.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Schematic of Sall4 alleles. (PDF 179 kb)

Supplementary Fig. 2

Patterning defects in Tbx5−/+ limbs. (PDF 425 kb)

Supplementary Fig. 3

Nuclear localization of Sall4 proteins. (PDF 307 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Koshiba-Takeuchi, K., Takeuchi, J., Arruda, E. et al. Cooperative and antagonistic interactions between Sall4 and Tbx5 pattern the mouse limb and heart. Nat Genet 38, 175–183 (2006). https://doi.org/10.1038/ng1707

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ng1707

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