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. 1988 Feb 1;106(2):461–467. doi: 10.1083/jcb.106.2.461

Changes in the expression of collagen genes show two stages in chondrocyte differentiation in vitro

PMCID: PMC2114986  PMID: 3339097

Abstract

This report deals with the quantitation of both mRNA and transcription activity of type I collagen gene and of three cartilage-specific collagens (types II, IX, and X) during in vitro differentiation of chick chondrocytes. Differentiation was obtained by transferal to suspension culture of dedifferentiated cells passaged for 3 wk as adherent cells. The type I collagen mRNA, highly represented in the dedifferentiated cells, rapidly decreased during chondrocyte differentiation. On the contrary, types II and IX collagen mRNAs sharply increased within the first week of suspension culture, peaked in the second week, and thereafter began to decrease. This decrease was particularly significant for type IX collagen mRNA. The level of type X collagen mRNA progressively increased during the course of the culture, reached its maximal value after 3-4 wk, and decreased only at a later stage of cell differentiation. As determined by in vitro run-off transcription assays, all these changes in collagen mRNA levels could be attributed to parallel modifications in the relative rate of transcription of the corresponding collagen genes. We suggest that chicken chondrocyte differentiation proceeds through at least two different steps: (a) first, transition from a stage characterized by a high level of type I collagen mRNA to a stage characterized by predominance of types II and IX collagen mRNAs; (b) later, transition to a stage characterized by the highest level of type X collagen mRNA.

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Selected References

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  1. Ambesi-Impiombato F. S., Parks L. A., Coon H. G. Culture of hormone-dependent functional epithelial cells from rat thyroids. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3455–3459. doi: 10.1073/pnas.77.6.3455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Capasso O., Tajana G., Cancedda R. Location of 64K collagen producer chondrocytes in developing chicken embryo tibiae. Mol Cell Biol. 1984 Jun;4(6):1163–1168. doi: 10.1128/mcb.4.6.1163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Castagnola P., Moro G., Descalzi-Cancedda F., Cancedda R. Type X collagen synthesis during in vitro development of chick embryo tibial chondrocytes. J Cell Biol. 1986 Jun;102(6):2310–2317. doi: 10.1083/jcb.102.6.2310. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Castagnola P., Torella G., Cancedda R. Type X collagen synthesis by cultured chondrocytes derived from the permanent cartilaginous region of chick embryo sternum. Dev Biol. 1987 Oct;123(2):332–337. doi: 10.1016/0012-1606(87)90391-5. [DOI] [PubMed] [Google Scholar]
  5. Clayton D. F., Darnell J. E., Jr Changes in liver-specific compared to common gene transcription during primary culture of mouse hepatocytes. Mol Cell Biol. 1983 Sep;3(9):1552–1561. doi: 10.1128/mcb.3.9.1552. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Duchêne M., Sobel M. E., Müller P. K. Levels of collagen mRNA in dedifferentiating chondrocytes. Exp Cell Res. 1982 Dec;142(2):317–324. doi: 10.1016/0014-4827(82)90373-1. [DOI] [PubMed] [Google Scholar]
  7. Focht R. J., Adams S. L. Tissue specificity of type I collagen gene expression is determined at both transcriptional and post-transcriptional levels. Mol Cell Biol. 1984 Sep;4(9):1843–1852. doi: 10.1128/mcb.4.9.1843. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kosher R. A., Kulyk W. M., Gay S. W. Collagen gene expression during limb cartilage differentiation. J Cell Biol. 1986 Apr;102(4):1151–1156. doi: 10.1083/jcb.102.4.1151. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kravis D., Upholt W. B. Quantitation of type II procollagen mRNA levels during chick limb cartilage differentiation. Dev Biol. 1985 Mar;108(1):164–172. doi: 10.1016/0012-1606(85)90018-1. [DOI] [PubMed] [Google Scholar]
  10. Lindell T. J., Weinberg F., Morris P. W., Roeder R. G., Rutter W. J. Specific inhibition of nuclear RNA polymerase II by alpha-amanitin. Science. 1970 Oct 23;170(3956):447–449. doi: 10.1126/science.170.3956.447. [DOI] [PubMed] [Google Scholar]
  11. McKnight G. S., Palmiter R. D. Transcriptional regulation of the ovalbumin and conalbumin genes by steroid hormones in chick oviduct. J Biol Chem. 1979 Sep 25;254(18):9050–9058. [PubMed] [Google Scholar]
  12. Müller-Glauser W., Humbel B., Glatt M., Sträuli P., Winterhalter K. H., Bruckner P. On the role of type IX collagen in the extracellular matrix of cartilage: type IX collagen is localized to intersections of collagen fibrils. J Cell Biol. 1986 May;102(5):1931–1939. doi: 10.1083/jcb.102.5.1931. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ninomiya Y., van der Rest M., Mayne R., Lozano G., Olsen B. R. Construction and characterization of cDNA encoding the alpha 2 chain of chicken type IX collagen. Biochemistry. 1985 Jul 16;24(15):4223–4229. doi: 10.1021/bi00336a061. [DOI] [PubMed] [Google Scholar]
  14. Saxe S. A., Lukens L. N., Pawlowski P. J. Changes in the nuclear and cytoplasmic levels of type I and type II collagen RNAs during growth of chondrocytes in 5-bromo-2'-deoxyuridine. J Biol Chem. 1985 Mar 25;260(6):3812–3819. [PubMed] [Google Scholar]
  15. Schmid T. M., Linsenmayer T. F. Immunohistochemical localization of short chain cartilage collagen (type X) in avian tissues. J Cell Biol. 1985 Feb;100(2):598–605. doi: 10.1083/jcb.100.2.598. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Solursh M., Jensen K. L., Reiter R. S., Schmid T. M., Linsenmayer T. F. Environmental regulation of type X collagen production by cultures of limb mesenchyme, mesectoderm, and sternal chondrocytes. Dev Biol. 1986 Sep;117(1):90–101. doi: 10.1016/0012-1606(86)90351-9. [DOI] [PubMed] [Google Scholar]
  17. Solursh M., Jensen K. L., Singley C. T., Linsenmayer T. F., Reiter R. S. Two distinct regulatory steps in cartilage differentiation. Dev Biol. 1982 Dec;94(2):311–325. doi: 10.1016/0012-1606(82)90350-5. [DOI] [PubMed] [Google Scholar]
  18. Tacchetti C., Quarto R., Nitsch L., Hartmann D. J., Cancedda R. In vitro morphogenesis of chick embryo hypertrophic cartilage. J Cell Biol. 1987 Aug;105(2):999–1006. doi: 10.1083/jcb.105.2.999. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Tullis R. H., Rubin H. Calcium protects DNase I from proteinase K: a new method for the removal of contaminating RNase from DNase I. Anal Biochem. 1980 Sep 1;107(1):260–264. doi: 10.1016/0003-2697(80)90519-9. [DOI] [PubMed] [Google Scholar]
  21. Yamamoto T., Sobel M. E., Adams S. L., Avvedimento V. E., DiLauro R., Pastan I., de Crombrugghe B., Showalter A., Pesciotta D., Fietzek P. Construction of a recombinant bacterial plasmid containing pro-alpha 1(I) collagen DNA sequences. J Biol Chem. 1980 Mar 25;255(6):2612–2615. [PubMed] [Google Scholar]
  22. Young M. F., Vogeli G., Nunez A. M., Fernandez M. P., Sullivan M., Sobel M. E. Isolation of cDNA and genomic DNA clones encoding type II collagen. Nucleic Acids Res. 1984 May 25;12(10):4207–4228. doi: 10.1093/nar/12.10.4207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. von der Mark K., von der Mark H. Immunological and biochemical studies of collagen type transition during in vitro chrondrogenesis of chick limb mesodermal cells. J Cell Biol. 1977 Jun;73(3):736–747. doi: 10.1083/jcb.73.3.736. [DOI] [PMC free article] [PubMed] [Google Scholar]

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