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Sox9 is required for cartilage formation


Chondrogenesis results in the formation of cartilages, initial skeletal elements that can serve as templates for endochondral bone formation. Cartilage formation begins with the condensation of mesenchyme cells followed by their differentiation into chondrocytes. Although much is known about the terminal differentiation products that are expressed by chondrocytes1,2,3, little is known about the factors that specify the chondrocyte lineage4,5,6. SOX9 is a high-mobility-group (HMG) domain transcription factor that is expressed in chondrocytes and other tissues7,8,9,10,11,12. In humans, SOX9 haploinsufficiency results in campomelic dysplasia, a lethal skeletal malformation syndrome, and XY sex reversal7,13,14,15,16. During embryogenesis, Sox9 is expressed in all cartilage primordia and cartilages, coincident with the expression of the collagen α1(II) gene (Col2a1; refs 8,11, 12). Sox9 is also expressed in other tissues, including the central nervous and urogenital systems8,9,10,11,12. Sox9 binds to essential sequences in the Col2a1 and collagen α2(XI) gene (Col11a2) chondrocyte-specific enhancers and can activate these enhancers in non-chondrocytic cells17,18,19. Here, Sox9 is identified as a regulator of the chondrocyte lineage. In mouse chimaeras, Sox9-/- cells are excluded from all cartilages but are present as a juxtaposed mesenchyme that does not express the chondrocyte-specific markers Col2a1, Col9a2, Col11a2 and Agc. This exclusion occurred cell autonomously at the condensing mesenchyme stage of chondrogenesis. Moreover, no cartilage developed in teratomas derived from Sox9-/- embryonic stem (ES) cells. Our results identify Sox9 as the first transcription factor that is essential for chondrocyte differentiation and cartilage formation.

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Figure 1: Generation of Sox9-mutant ES cell lines.
Figure 2: Sox9 chimaeras at 15.5 dpc.
Figure 3: Sox9+/- (a-c) and Sox9-/- chimaeras (d-k) at 15.5 dpc.
Figure 4: Sox9 chimaeras at 11.5 and 12.5 dpc.
Figure 5: Teratomas derived from wild-type, Sox9+/- and Sox9-/- ES cell lines.


  1. Stockwell, R.A. in Biology of Cartilage Cells (Cambridge University Press, Cambridge, UK, 1979).

    Google Scholar 

  2. Hall, B. & Newman, S. in Cartilage: Molecular Aspects (CRC Press, Boca Raton, Florida, 1991).

    Google Scholar 

  3. Mundlos, S. & Olsen, B.R. Heritable diseases of the skeleton. Part I: molecular insights into skeletal development-transcription factors and signaling pathways. FASEB J. 11, 125 –132 (1997).

    Article  CAS  Google Scholar 

  4. Cancedda, R., Descalzi Cancedda, F. & Castagnola, P. Chondrocyte differentiation. Int. Rev. Cytol. 159, 265–358 (1995).

    Article  CAS  Google Scholar 

  5. Erlebacher, A., Filvaroff, E.H., Gitelman, S.E. & Derynck, R. Toward a molecular understanding of skeletal development. Cell 80, 371–378 ( 1995).

    Article  CAS  Google Scholar 

  6. Mundlos, S. & Olsen, B.R. Heritable diseases of the skeleton. Part II: molecular insights into skeletal development-matrix components and their homeostasis. FASEB J. 11, 227– 233 (1997).

    Article  CAS  Google Scholar 

  7. Wagner, T. et al. Autosomal sex reversal and campomelic dysplasia are caused by mutations in and around the SRY-related gene SOX9. Cell 79, 1111–1120 ( 1994).

    Article  CAS  Google Scholar 

  8. Wright, E. et al. The Sry-related gene Sox-9 is expressed during chrondrogenesis in mouse embryos. Nature Genet. 9, 15–20 (1995).

    Article  CAS  Google Scholar 

  9. Kent, J., Wheatley, S.C., Andrews, J.E., Sinclair, A.H. & Koopman, P. A male-specific role for SOX9 in vertebrate sex determination. Development 122 , 2813–2822 (1996).

    CAS  PubMed  Google Scholar 

  10. Morais da Silva, S. et al. Sox9 expression during gonadal development implies a conserved role for the gene in testis differentiation in mammals and birds. Nature Genet. 14, 62–68 (1996).

    Article  CAS  Google Scholar 

  11. Ng, L.-J. et al. Sox9 binds DNA, activates transcription, and coexpresses with type II collagen during chondrogenesis in the mouse. Dev. Biol. 183, 108–121 ( 1997).

    Article  CAS  Google Scholar 

  12. Zhao, Q., Eberspaecher, H., Lefebvre, V. & de Crombrugghe, B. Parallel expression of Sox9 and Col2a1 in cells undergoing chondrogenesis. Dev. Dyn. 209, 377–386 (1997).

    Article  CAS  Google Scholar 

  13. Hovmoller, M.L. et al. Campomelic dwarfism. A genetically determined mesenchymal disorder combined with sex reversal. Hereditas 86, 51–62 (1977).

    Article  CAS  Google Scholar 

  14. Takahashi, H., Sando, I. & Masutani, H. Temporal bone histopathological findings in campomelic dysplasia. J. Laryngol. Otol. 106, 361– 365 (1992).

    Article  CAS  Google Scholar 

  15. Foster, J.W. et al. Campomelic dysplasia and autosomal sex reversal caused by mutations in an SRY-related gene. Nature 372 , 525–530 (1994).

    Article  CAS  Google Scholar 

  16. Kwok, C. et al. Mutations in SOX9, the gene responsible for campomelic dysplasia and autosomal sex reversal. Am. J. Hum. Genet. 57, 1028–1036 (1995).

    CAS  PubMed  PubMed Central  Google Scholar 

  17. Bell, D.M. et al. Sox9 directly regulates the type-II collagen gene. Nature Genet. 16, 174–178 (1997).

    Article  CAS  Google Scholar 

  18. Lefebvre, V., Huang, W., Harley, V.R., Goodfellow, P.N. & de Crombrugghe, B. SOX9 is a potent activator of the chondrocyte-specific enhancer of the proα1(II) collagen gene. Mol. Cell. Biol. 17, 2336–2346 ( 1997).

    Article  CAS  Google Scholar 

  19. Bridgewater, L.C., Lefebvre, V. & de Crombrugghe, B. Chondrocyte-specific enhancer elements in the Col11a2 gene resemble the Col2a1 tissue-specific enhancer. J. Biol. Chem. 273, 14998–15006 (1998).

    Article  CAS  Google Scholar 

  20. Widelitz, R.B., Jiang, T.X., Murray, B.A. & Chuong, C.M. Adhesion molecules in skeletogenesis: II. Neural cell adhesion molecules mediate precartilaginous mesenchymal condensations and enhance chondrogenesis. J. Cell Physiol. 156, 399–411 (1993).

    Article  CAS  Google Scholar 

  21. Oberlender, S.A. & Tuan, R.S. Expression and functional involvement of N-cadherin in embryonic limb chondrogenesis. Development 120, 177–187 (1994).

    CAS  PubMed  Google Scholar 

  22. Mountford, P. et al. Dicistronic targeting constructs: reporters and modifiers of mammalian gene expression. Proc. Natl Acad. Sci. USA 91, 4303–4307 (1994).

    Article  CAS  Google Scholar 

  23. Behringer, R.R., Finegold, M.J. & Cate, R.L. Müllerian-inhibiting substance function during mammalian sexual development. Cell 79, 415 –425 (1994).

    Article  CAS  Google Scholar 

  24. Abuin, A. & Bradley, A. Recycling selectable markers in mouse embryonic stem cells. Mol. Cell. Biol. 16, 1851–1856 (1996).

    Article  CAS  Google Scholar 

  25. Hogan, B., Beddington, R., Costantini, F. & Lacy, E. in Manipulating the Mouse Embryo: A Laboratory Manual (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 1994).

    Google Scholar 

  26. Wilkinson, D.G. & Nieto, M.A. Detection of messenger RNA by in situ hybridization to tissue sections and whole-mounts. Methods Enzymol. 225, 361–373 (1993).

    Article  CAS  Google Scholar 

  27. Metsäranta, M., Toman, D., de Crombrugghe, B. & Vuorio, E. Specific hybridization probes for mouse type I, II, III and IX collagen mRNAs. Biochim. Biophys. Acta 1089, 241– 243 (1991).

    Article  Google Scholar 

  28. Glumoff, V., Savontaus, M., Vehanen, J. & Vuorio, E. Analysis of aggrecan and tenascin gene expression in mouse skeletal tissues by northern and in situ hybridization using species specific cDNA probes. Biochim. Biophys. Acta 1219, 613– 622 (1994).

    Article  Google Scholar 

  29. Elima, K. et al. Specific hybridization probes for mouse α2(IX) and α1(X) collagen mRNAs. Biochim. Biophys. Acta 1130, 78–80 (1992).

    Article  CAS  Google Scholar 

  30. Robertson, E.J. in Teratocarcinomas and Embryonic Stem Cells: A Practical Approach (IRL Press, Oxford, UK, 1987).

    Google Scholar 

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We thank A. Bradley for AB-1 ES and SNL 76/7 STO cell lines; M. Wakamiya for IRES-lacZ-pA loxP-flanked PGKneobpA plasmid; E. Vuorio and L. Bridgewater for RNA in situ hybridization probes; D. Whitworth for assistance with embryos; H. Eberspaecher for advice on histology; G. Pinero for histological assistance; and A. Bradley, R. Johnson and V. Lefebvre for helpful comments on the manuscript. The anti-collagen II monoclonal antibody developed by T. Linsenmayer was obtained from the Developmental Studies Hybridoma Bank. This study was supported by NIH grants HD30284 to R.R.B. and AR42919 to R.R.B. and B. de C.

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Correspondence to Richard R. Behringer or Benoit de Crombrugghe.

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Bi, W., Deng, J., Zhang, Z. et al. Sox9 is required for cartilage formation. Nat Genet 22, 85–89 (1999).

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