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Sox9 is required for cartilage formation

Nature Genetics volume 22, pages 85–89 (1999)Cite this article

Abstract

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.

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Acknowledgements

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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Authors and Affiliations

  1. Department of Molecular Genetics, The University of Texas, M.D. Anderson Cancer Center, Houston, 77030, Texas, USA

    Weimin Bi, Jian Min Deng, Zhaoping Zhang, Richard R. Behringer & Benoit de Crombrugghe

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  1. Weimin Bi
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  2. Jian Min Deng
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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). https://doi.org/10.1038/8792

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