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. 1994 Dec 2;127(6):2093–2101. doi: 10.1083/jcb.127.6.2093

Tenascin-C expression by fibroblasts is elevated in stressed collagen gels

PMCID: PMC2120287  PMID: 7528751

Abstract

Chick embryo fibroblasts cultured on a collagen matrix exert tractional forces leading to the contraction of unrestrained, floating collagen gels and to the development of tension in attached, restrained gels. On a restrained, attached collagen gel the fibroblasts synthesize large quantities of tenascin-C, whereas in a floating, contracting gel tenascin-C synthesis is decreased. This regulation of tenascin-C synthesis can be observed by the secretion of metabolically labeled tenascin-C into the conditioned medium, as well as by the deposition of tenascin-C into the collagen matrix as judged by immunofluorescence. Regulation appears to occur at the transcriptional level, because when cells on attached or floating collagen gels are transfected with promoter constructs of the tenascin-C gene, luciferase expression driven by the tenascin-C promoter parallels the effects measured for endogenous tenascin-C synthesis, whereas luciferase expression under the control of the SV40 promoter does not depend on the state of the collagen gel. The promoter region responsible for tenascin-C induction on attached collagen gels is distinct from the region important for the induction of tenascin-C by serum, and may define a novel kind of response element. By joining this tenascin-C sequence to the SV40 promoter of a reporter plasmid, its activity can be transferred to the heterologous promoter. We propose that the tenascin-C promoter is directly or indirectly activated in fibroblasts generating and experiencing mechanical stress within a restrained collagen matrix. This may be an important aspect of the regulation of tenascin-C expression during embryogenesis as well as during wound healing and other regenerative and morphogenetic processes.

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Selected References

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  1. Adams J. C., Watt F. M. Regulation of development and differentiation by the extracellular matrix. Development. 1993 Apr;117(4):1183–1198. doi: 10.1242/dev.117.4.1183. [DOI] [PubMed] [Google Scholar]
  2. Artelt P., Grannemann R., Stocking C., Friel J., Bartsch J., Hauser H. The prokaryotic neomycin-resistance-encoding gene acts as a transcriptional silencer in eukaryotic cells. Gene. 1991 Mar 15;99(2):249–254. doi: 10.1016/0378-1119(91)90134-w. [DOI] [PubMed] [Google Scholar]
  3. Bell E., Ivarsson B., Merrill C. Production of a tissue-like structure by contraction of collagen lattices by human fibroblasts of different proliferative potential in vitro. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1274–1278. doi: 10.1073/pnas.76.3.1274. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Ben-Ze'ev A. Animal cell shape changes and gene expression. Bioessays. 1991 May;13(5):207–212. doi: 10.1002/bies.950130502. [DOI] [PubMed] [Google Scholar]
  5. Bissell M. J., Hall H. G., Parry G. How does the extracellular matrix direct gene expression? J Theor Biol. 1982 Nov 7;99(1):31–68. doi: 10.1016/0022-5193(82)90388-5. [DOI] [PubMed] [Google Scholar]
  6. Chan B. M., Kassner P. D., Schiro J. A., Byers H. R., Kupper T. S., Hemler M. E. Distinct cellular functions mediated by different VLA integrin alpha subunit cytoplasmic domains. Cell. 1992 Mar 20;68(6):1051–1060. doi: 10.1016/0092-8674(92)90077-p. [DOI] [PubMed] [Google Scholar]
  7. Chiquet-Ehrismann R., Kalla P., Pearson C. A. Participation of tenascin and transforming growth factor-beta in reciprocal epithelial-mesenchymal interactions of MCF7 cells and fibroblasts. Cancer Res. 1989 Aug 1;49(15):4322–4325. [PubMed] [Google Scholar]
  8. Chiquet-Ehrismann R., Mackie E. J., Pearson C. A., Sakakura T. Tenascin: an extracellular matrix protein involved in tissue interactions during fetal development and oncogenesis. Cell. 1986 Oct 10;47(1):131–139. doi: 10.1016/0092-8674(86)90374-0. [DOI] [PubMed] [Google Scholar]
  9. Chiquet-Ehrismann R. Tenascin and other adhesion-modulating proteins in cancer. Semin Cancer Biol. 1993 Oct;4(5):301–310. [PubMed] [Google Scholar]
  10. Chiquet M., Vrucinić-Filipi N., Schenk S., Beck K., Chiquet-Ehrismann R. Isolation of chick tenascin variants and fragments. A C-terminal heparin-binding fragment produced by cleavage of the extra domain from the largest subunit splicing variant. Eur J Biochem. 1991 Jul 15;199(2):379–388. doi: 10.1111/j.1432-1033.1991.tb16134.x. [DOI] [PubMed] [Google Scholar]
  11. Delvoye P., Wiliquet P., Levêque J. L., Nusgens B. V., Lapière C. M. Measurement of mechanical forces generated by skin fibroblasts embedded in a three-dimensional collagen gel. J Invest Dermatol. 1991 Nov;97(5):898–902. doi: 10.1111/1523-1747.ep12491651. [DOI] [PubMed] [Google Scholar]
  12. Ekblom M., Fässler R., Tomasini-Johansson B., Nilsson K., Ekblom P. Downregulation of tenascin expression by glucocorticoids in bone marrow stromal cells and in fibroblasts. J Cell Biol. 1993 Nov;123(4):1037–1045. doi: 10.1083/jcb.123.4.1037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Erickson H. P., Bourdon M. A. Tenascin: an extracellular matrix protein prominent in specialized embryonic tissues and tumors. Annu Rev Cell Biol. 1989;5:71–92. doi: 10.1146/annurev.cb.05.110189.000443. [DOI] [PubMed] [Google Scholar]
  14. Gullberg D., Tingström A., Thuresson A. C., Olsson L., Terracio L., Borg T. K., Rubin K. Beta 1 integrin-mediated collagen gel contraction is stimulated by PDGF. Exp Cell Res. 1990 Feb;186(2):264–272. doi: 10.1016/0014-4827(90)90305-t. [DOI] [PubMed] [Google Scholar]
  15. Haskill S., Beg A. A., Tompkins S. M., Morris J. S., Yurochko A. D., Sampson-Johannes A., Mondal K., Ralph P., Baldwin A. S., Jr Characterization of an immediate-early gene induced in adherent monocytes that encodes I kappa B-like activity. Cell. 1991 Jun 28;65(7):1281–1289. doi: 10.1016/0092-8674(91)90022-q. [DOI] [PubMed] [Google Scholar]
  16. Hynes R. O. Integrins: versatility, modulation, and signaling in cell adhesion. Cell. 1992 Apr 3;69(1):11–25. doi: 10.1016/0092-8674(92)90115-s. [DOI] [PubMed] [Google Scholar]
  17. Ingber D. E. The riddle of morphogenesis: a question of solution chemistry or molecular cell engineering? Cell. 1993 Dec 31;75(7):1249–1252. doi: 10.1016/0092-8674(93)90612-t. [DOI] [PubMed] [Google Scholar]
  18. Jones F. S., Chalepakis G., Gruss P., Edelman G. M. Activation of the cytotactin promoter by the homeobox-containing gene Evx-1. Proc Natl Acad Sci U S A. 1992 Mar 15;89(6):2091–2095. doi: 10.1073/pnas.89.6.2091. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Jones F. S., Crossin K. L., Cunningham B. A., Edelman G. M. Identification and characterization of the promoter for the cytotactin gene. Proc Natl Acad Sci U S A. 1990 Sep;87(17):6497–6501. doi: 10.1073/pnas.87.17.6497. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Juliano R. L., Haskill S. Signal transduction from the extracellular matrix. J Cell Biol. 1993 Feb;120(3):577–585. doi: 10.1083/jcb.120.3.577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kim S. J., Denhez F., Kim K. Y., Holt J. T., Sporn M. B., Roberts A. B. Activation of the second promoter of the transforming growth factor-beta 1 gene by transforming growth factor-beta 1 and phorbol ester occurs through the same target sequences. J Biol Chem. 1989 Nov 15;264(32):19373–19378. [PubMed] [Google Scholar]
  22. Klein C. E., Dressel D., Steinmayer T., Mauch C., Eckes B., Krieg T., Bankert R. B., Weber L. Integrin alpha 2 beta 1 is upregulated in fibroblasts and highly aggressive melanoma cells in three-dimensional collagen lattices and mediates the reorganization of collagen I fibrils. J Cell Biol. 1991 Dec;115(5):1427–1436. doi: 10.1083/jcb.115.5.1427. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Klemsz M. J., McKercher S. R., Celada A., Van Beveren C., Maki R. A. The macrophage and B cell-specific transcription factor PU.1 is related to the ets oncogene. Cell. 1990 Apr 6;61(1):113–124. doi: 10.1016/0092-8674(90)90219-5. [DOI] [PubMed] [Google Scholar]
  24. Koch M., Bernasconi C., Chiquet M. A major oligomeric fibroblast proteoglycan identified as a novel large form of type-XII collagen. Eur J Biochem. 1992 Aug 1;207(3):847–856. doi: 10.1111/j.1432-1033.1992.tb17116.x. [DOI] [PubMed] [Google Scholar]
  25. Kolodney M. S., Wysolmerski R. B. Isometric contraction by fibroblasts and endothelial cells in tissue culture: a quantitative study. J Cell Biol. 1992 Apr;117(1):73–82. doi: 10.1083/jcb.117.1.73. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Komuro I., Katoh Y., Kaida T., Shibazaki Y., Kurabayashi M., Hoh E., Takaku F., Yazaki Y. Mechanical loading stimulates cell hypertrophy and specific gene expression in cultured rat cardiac myocytes. Possible role of protein kinase C activation. J Biol Chem. 1991 Jan 15;266(2):1265–1268. [PubMed] [Google Scholar]
  27. Liu J. K., DiPersio C. M., Zaret K. S. Extracellular signals that regulate liver transcription factors during hepatic differentiation in vitro. Mol Cell Biol. 1991 Feb;11(2):773–784. doi: 10.1128/mcb.11.2.773. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Lux S. E., John K. M., Bennett V. Analysis of cDNA for human erythrocyte ankyrin indicates a repeated structure with homology to tissue-differentiation and cell-cycle control proteins. Nature. 1990 Mar 1;344(6261):36–42. doi: 10.1038/344036a0. [DOI] [PubMed] [Google Scholar]
  29. Mackie E. J., Scott-Burden T., Hahn A. W., Kern F., Bernhardt J., Regenass S., Weller A., Bühler F. R. Expression of tenascin by vascular smooth muscle cells. Alterations in hypertensive rats and stimulation by angiotensin II. Am J Pathol. 1992 Aug;141(2):377–388. [PMC free article] [PubMed] [Google Scholar]
  30. Mackie E. J., Thesleff I., Chiquet-Ehrismann R. Tenascin is associated with chondrogenic and osteogenic differentiation in vivo and promotes chondrogenesis in vitro. J Cell Biol. 1987 Dec;105(6 Pt 1):2569–2579. doi: 10.1083/jcb.105.6.2569. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Massagué J. The transforming growth factor-beta family. Annu Rev Cell Biol. 1990;6:597–641. doi: 10.1146/annurev.cb.06.110190.003121. [DOI] [PubMed] [Google Scholar]
  32. Mitchell P. J., Tjian R. Transcriptional regulation in mammalian cells by sequence-specific DNA binding proteins. Science. 1989 Jul 28;245(4916):371–378. doi: 10.1126/science.2667136. [DOI] [PubMed] [Google Scholar]
  33. Mochitate K., Pawelek P., Grinnell F. Stress relaxation of contracted collagen gels: disruption of actin filament bundles, release of cell surface fibronectin, and down-regulation of DNA and protein synthesis. Exp Cell Res. 1991 Mar;193(1):198–207. doi: 10.1016/0014-4827(91)90556-a. [DOI] [PubMed] [Google Scholar]
  34. Pearson C. A., Pearson D., Shibahara S., Hofsteenge J., Chiquet-Ehrismann R. Tenascin: cDNA cloning and induction by TGF-beta. EMBO J. 1988 Oct;7(10):2977–2982. doi: 10.1002/j.1460-2075.1988.tb03160.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Prieto A. L., Jones F. S., Cunningham B. A., Crossin K. L., Edelman G. M. Localization during development of alternatively spliced forms of cytotactin mRNA by in situ hybridization. J Cell Biol. 1990 Aug;111(2):685–698. doi: 10.1083/jcb.111.2.685. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Resnick N., Collins T., Atkinson W., Bonthron D. T., Dewey C. F., Jr, Gimbrone M. A., Jr Platelet-derived growth factor B chain promoter contains a cis-acting fluid shear-stress-responsive element. Proc Natl Acad Sci U S A. 1993 May 15;90(10):4591–4595. doi: 10.1073/pnas.90.10.4591. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Rettig W. J., Erickson H. P., Albino A. P., Garin-Chesa P. Induction of human tenascin (neuronectin) by growth factors and cytokines: cell type-specific signals and signalling pathways. J Cell Sci. 1994 Feb;107(Pt 2):487–497. [PubMed] [Google Scholar]
  38. Risse G., Jooss K., Neuberg M., Brüller H. J., Müller R. Asymmetrical recognition of the palindromic AP1 binding site (TRE) by Fos protein complexes. EMBO J. 1989 Dec 1;8(12):3825–3832. doi: 10.1002/j.1460-2075.1989.tb08560.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Rossi P., Karsenty G., Roberts A. B., Roche N. S., Sporn M. B., de Crombrugghe B. A nuclear factor 1 binding site mediates the transcriptional activation of a type I collagen promoter by transforming growth factor-beta. Cell. 1988 Feb 12;52(3):405–414. doi: 10.1016/s0092-8674(88)80033-3. [DOI] [PubMed] [Google Scholar]
  40. Sadler I., Crawford A. W., Michelsen J. W., Beckerle M. C. Zyxin and cCRP: two interactive LIM domain proteins associated with the cytoskeleton. J Cell Biol. 1992 Dec;119(6):1573–1587. doi: 10.1083/jcb.119.6.1573. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Schmidhauser C., Casperson G. F., Myers C. A., Sanzo K. T., Bolten S., Bissell M. J. A novel transcriptional enhancer is involved in the prolactin- and extracellular matrix-dependent regulation of beta-casein gene expression. Mol Biol Cell. 1992 Jun;3(6):699–709. doi: 10.1091/mbc.3.6.699. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Schwartz M. A. Transmembrane signalling by integrins. Trends Cell Biol. 1992 Oct;2(10):304–308. doi: 10.1016/0962-8924(92)90120-c. [DOI] [PubMed] [Google Scholar]
  43. Stopak D., Harris A. K. Connective tissue morphogenesis by fibroblast traction. I. Tissue culture observations. Dev Biol. 1982 Apr;90(2):383–398. doi: 10.1016/0012-1606(82)90388-8. [DOI] [PubMed] [Google Scholar]
  44. Tucker R. P., Hammarback J. A., Jenrath D. A., Mackie E. J., Xu Y. Tenascin expression in the mouse: in situ localization and induction in vitro by bFGF. J Cell Sci. 1993 Jan;104(Pt 1):69–76. doi: 10.1242/jcs.104.1.69. [DOI] [PubMed] [Google Scholar]
  45. Umbhauer M., Riou J. F., Spring J., Smith J. C., Boucaut J. C. Expression of tenascin mRNA in mesoderm during Xenopus laevis embryogenesis: the potential role of mesoderm patterning in tenascin regionalization. Development. 1992 Sep;116(1):147–157. doi: 10.1242/dev.116.1.147. [DOI] [PubMed] [Google Scholar]
  46. Van Obberghen-Schilling E., Roche N. S., Flanders K. C., Sporn M. B., Roberts A. B. Transforming growth factor beta 1 positively regulates its own expression in normal and transformed cells. J Biol Chem. 1988 Jun 5;263(16):7741–7746. [PubMed] [Google Scholar]
  47. Vrucinić-Filipi N., Chiquet-Ehrismann R. Tenascin function and regulation of expression. Symp Soc Exp Biol. 1993;47:155–162. [PubMed] [Google Scholar]
  48. Wang N., Butler J. P., Ingber D. E. Mechanotransduction across the cell surface and through the cytoskeleton. Science. 1993 May 21;260(5111):1124–1127. doi: 10.1126/science.7684161. [DOI] [PubMed] [Google Scholar]
  49. Wilson E., Mai Q., Sudhir K., Weiss R. H., Ives H. E. Mechanical strain induces growth of vascular smooth muscle cells via autocrine action of PDGF. J Cell Biol. 1993 Nov;123(3):741–747. doi: 10.1083/jcb.123.3.741. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Yurochko A. D., Liu D. Y., Eierman D., Haskill S. Integrins as a primary signal transduction molecule regulating monocyte immediate-early gene induction. Proc Natl Acad Sci U S A. 1992 Oct 1;89(19):9034–9038. doi: 10.1073/pnas.89.19.9034. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Zachary I., Rozengurt E. Focal adhesion kinase (p125FAK): a point of convergence in the action of neuropeptides, integrins, and oncogenes. Cell. 1992 Dec 11;71(6):891–894. doi: 10.1016/0092-8674(92)90385-p. [DOI] [PubMed] [Google Scholar]
  52. von der Ahe D., Janich S., Scheidereit C., Renkawitz R., Schütz G., Beato M. Glucocorticoid and progesterone receptors bind to the same sites in two hormonally regulated promoters. Nature. 1985 Feb 21;313(6004):706–709. doi: 10.1038/313706a0. [DOI] [PubMed] [Google Scholar]

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