Skip to main content
NCBI home page
Journal of Anatomy logoLink to Journal of Anatomy
. 1997 Jun;190(Pt 4):523–532. doi: 10.1046/j.1469-7580.1997.19040523.x

Structural colocalisation of type VI collagen and fibronectin in agarose cultured chondrocytes and isolated chondrons extracted from adult canine tibial cartilage

JIANG CHANG 1 , HIDETO NAKAJIMA 2 , C ANTHONY POOLE 1
PMCID: PMC1467638  PMID: 9183676

Abstract

Cell–matrix and matrix–matrix interactions are of critical importance in regulating the development, maintenance and repair of articular cartilage. In this study, we examined the structural colocalisation of type VI collagen and fibronectin in isolated chondrons and long-term agarose cultured chondrocytes extracted from normal adult canine articular cartilage. Using double labelling immunohistochemistry in conjunction with dual channel confocal microscopy and digital image processing we demonstrate that type VI collagen and fibronectin are distributed in a similar staining pattern and are colocalised at the surface of cultured chondrocytes and isolated chondrons. The results suggest that type VI collagen and fibronectin may play a role in both cell–matrix adhesion and matrix–matrix cohesion in the pericellular microenvironment surrounding articular cartilage chondrocytes.

Keywords: Articular cartilage, dog, extracellular matrix, matrix–matrix, confocal microscopy

Full Text

The Full Text of this article is available as a PDF (452.3 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Akiyama S. K., Nagata K., Yamada K. M. Cell surface receptors for extracellular matrix components. Biochim Biophys Acta. 1990 Feb 28;1031(1):91–110. doi: 10.1016/0304-4157(90)90004-v. [DOI] [PubMed] [Google Scholar]
  2. Amenta P. S., Gay S., Vaheri A., Martinez-Hernandez A. The extracellular matrix is an integrated unit: ultrastructural localization of collagen types I, III, IV, V, VI, fibronectin, and laminin in human term placenta. Coll Relat Res. 1986 Jun;6(2):125–152. doi: 10.1016/s0174-173x(86)80021-8. [DOI] [PubMed] [Google Scholar]
  3. Ayad S., Evans H., Weiss J. B., Holt L. Type VI collagen but not type V collagen is present in cartilage. Coll Relat Res. 1984 Mar;4(2):165–168. doi: 10.1016/s0174-173x(84)80023-0. [DOI] [PubMed] [Google Scholar]
  4. Aydelotte M. B., Kuettner K. E. Differences between sub-populations of cultured bovine articular chondrocytes. I. Morphology and cartilage matrix production. Connect Tissue Res. 1988;18(3):205–222. doi: 10.3109/03008208809016808. [DOI] [PubMed] [Google Scholar]
  5. Bidanset D. J., Guidry C., Rosenberg L. C., Choi H. U., Timpl R., Hook M. Binding of the proteoglycan decorin to collagen type VI. J Biol Chem. 1992 Mar 15;267(8):5250–5256. [PubMed] [Google Scholar]
  6. Chevalier X. Fibronectin, cartilage, and osteoarthritis. Semin Arthritis Rheum. 1993 Apr;22(5):307–318. doi: 10.1016/s0049-0172(05)80010-1. [DOI] [PubMed] [Google Scholar]
  7. Doane K. J., Yang G., Birk D. E. Corneal cell-matrix interactions: type VI collagen promotes adhesion and spreading of corneal fibroblasts. Exp Cell Res. 1992 Jun;200(2):490–499. doi: 10.1016/0014-4827(92)90200-r. [DOI] [PubMed] [Google Scholar]
  8. Engel J., Furthmayr H., Odermatt E., von der Mark H., Aumailley M., Fleischmajer R., Timpl R. Structure and macromolecular organization of type VI collagen. Ann N Y Acad Sci. 1985;460:25–37. doi: 10.1111/j.1749-6632.1985.tb51154.x. [DOI] [PubMed] [Google Scholar]
  9. Evans H. B., Ayad S., Abedin M. Z., Hopkins S., Morgan K., Walton K. W., Weiss J. B., Holt P. J. Localisation of collagen types and fibronectin in cartilage by immunofluorescence. Ann Rheum Dis. 1983 Oct;42(5):575–581. doi: 10.1136/ard.42.5.575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Glant T. T., Hadházy C., Mikecz K., Sipos A. Appearance and persistence of fibronectin in cartilage. Specific interaction of fibronectin with collagen type II. Histochemistry. 1985;82(2):149–158. doi: 10.1007/BF00708199. [DOI] [PubMed] [Google Scholar]
  11. Hirsch M. S., Cook S. C., Killiany R., Hartford Svoboda K. K. Increased cell diameter precedes chondrocyte terminal differentiation, whereas cell-matrix attachment complex proteins appear constant. Anat Rec. 1996 Mar;244(3):284–296. doi: 10.1002/(SICI)1097-0185(199603)244:3<284::AID-AR2>3.0.CO;2-Z. [DOI] [PubMed] [Google Scholar]
  12. Hynes R. O. Integrins: a family of cell surface receptors. Cell. 1987 Feb 27;48(4):549–554. doi: 10.1016/0092-8674(87)90233-9. [DOI] [PubMed] [Google Scholar]
  13. Keene D. R., Engvall E., Glanville R. W. Ultrastructure of type VI collagen in human skin and cartilage suggests an anchoring function for this filamentous network. J Cell Biol. 1988 Nov;107(5):1995–2006. doi: 10.1083/jcb.107.5.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kielty C. M., Boot-Handford R. P., Ayad S., Shuttleworth C. A., Grant M. E. Molecular composition of type VI collagen. Evidence for chain heterogeneity in mammalian tissues and cultured cells. Biochem J. 1990 Dec 15;272(3):787–795. doi: 10.1042/bj2720787. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kielty C. M., Whittaker S. P., Grant M. E., Shuttleworth C. A. Type VI collagen microfibrils: evidence for a structural association with hyaluronan. J Cell Biol. 1992 Aug;118(4):979–990. doi: 10.1083/jcb.118.4.979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kirsch T., Pfäffle M. Selective binding of anchorin CII (annexin V) to type II and X collagen and to chondrocalcin (C-propeptide of type II collagen). Implications for anchoring function between matrix vesicles and matrix proteins. FEBS Lett. 1992 Sep 28;310(2):143–147. doi: 10.1016/0014-5793(92)81316-e. [DOI] [PubMed] [Google Scholar]
  17. Knudson C. B. Hyaluronan receptor-directed assembly of chondrocyte pericellular matrix. J Cell Biol. 1993 Feb;120(3):825–834. doi: 10.1083/jcb.120.3.825. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lee G. M., Johnstone B., Jacobson K., Caterson B. The dynamic structure of the pericellular matrix on living cells. J Cell Biol. 1993 Dec;123(6 Pt 2):1899–1907. doi: 10.1083/jcb.123.6.1899. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. McComb R. D., Moul J. M., Bigner D. D. Distribution of type VI collagen in human gliomas: comparison with fibronectin and glioma-mesenchymal matrix glycoprotein. J Neuropathol Exp Neurol. 1987 Nov;46(6):623–633. doi: 10.1097/00005072-198711000-00002. [DOI] [PubMed] [Google Scholar]
  20. McDevitt C. A., Marcelino J., Tucker L. Interaction of intact type VI collagen with hyaluronan. FEBS Lett. 1991 Dec 9;294(3):167–170. doi: 10.1016/0014-5793(91)80660-u. [DOI] [PubMed] [Google Scholar]
  21. McDevitt C. A., Pahl J. A., Ayad S., Miller R. R., Uratsuji M., Andrish J. T. Experimental osteoarthritic articular cartilage is enriched in guanidine soluble type VI collagen. Biochem Biophys Res Commun. 1988 Nov 30;157(1):250–255. doi: 10.1016/s0006-291x(88)80040-8. [DOI] [PubMed] [Google Scholar]
  22. Poole C. A., Ayad S., Gilbert R. T. Chondrons from articular cartilage. V. Immunohistochemical evaluation of type VI collagen organisation in isolated chondrons by light, confocal and electron microscopy. J Cell Sci. 1992 Dec;103(Pt 4):1101–1110. doi: 10.1242/jcs.103.4.1101. [DOI] [PubMed] [Google Scholar]
  23. Poole C. A., Ayad S., Schofield J. R. Chondrons from articular cartilage: I. Immunolocalization of type VI collagen in the pericellular capsule of isolated canine tibial chondrons. J Cell Sci. 1988 Aug;90(Pt 4):635–643. doi: 10.1242/jcs.90.4.635. [DOI] [PubMed] [Google Scholar]
  24. Poole C. A., Brookes N. H., Gilbert R. T., Beaumont B. W., Crowther A., Scott L., Merrilees M. J. Detection of viable and non-viable cells in connective tissue explants using the fixable fluoroprobes 5-chloromethylfluorescein diacetate and ethidium homodimer-1. Connect Tissue Res. 1996;33(4):233–241. doi: 10.3109/03008209609028880. [DOI] [PubMed] [Google Scholar]
  25. Poole C. A., Flint M. H., Beaumont B. W. Chondrons extracted from canine tibial cartilage: preliminary report on their isolation and structure. J Orthop Res. 1988;6(3):408–419. doi: 10.1002/jor.1100060312. [DOI] [PubMed] [Google Scholar]
  26. Poole C. A., Flint M. H., Beaumont B. W. Chondrons in cartilage: ultrastructural analysis of the pericellular microenvironment in adult human articular cartilages. J Orthop Res. 1987;5(4):509–522. doi: 10.1002/jor.1100050406. [DOI] [PubMed] [Google Scholar]
  27. Poole C. A., Glant T. T., Schofield J. R. Chondrons from articular cartilage. (IV). Immunolocalization of proteoglycan epitopes in isolated canine tibial chondrons. J Histochem Cytochem. 1991 Sep;39(9):1175–1187. doi: 10.1177/39.9.1717545. [DOI] [PubMed] [Google Scholar]
  28. Poole C. A., Honda T., Skinner S. J., Schofield J. R., Hyde K. F., Shinkai H. Chondrons from articular cartilage (II): Analysis of the glycosaminoglycans in the cellular microenvironment of isolated canine chondrons. Connect Tissue Res. 1990;24(3-4):319–330. doi: 10.3109/03008209009152158. [DOI] [PubMed] [Google Scholar]
  29. Poole C. A., Wotton S. F., Duance V. C. Localization of type IX collagen in chondrons isolated from porcine articular cartilage and rat chondrosarcoma. Histochem J. 1988 Oct;20(10):567–574. doi: 10.1007/BF01002611. [DOI] [PubMed] [Google Scholar]
  30. Ratcliffe A., Fryer P. R., Hardingham T. E. The distribution of aggregating proteoglycans in articular cartilage: comparison of quantitative immunoelectron microscopy with radioimmunoassay and biochemical analysis. J Histochem Cytochem. 1984 Feb;32(2):193–201. doi: 10.1177/32.2.6363519. [DOI] [PubMed] [Google Scholar]
  31. Roberts S., Ayad S., Menage P. J. Immunolocalisation of type VI collagen in the intervertebral disc. Ann Rheum Dis. 1991 Nov;50(11):787–791. doi: 10.1136/ard.50.11.787. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Ronzière M. C., Ricard-Blum S., Tiollier J., Hartmann D. J., Garrone R., Herbage D. Comparative analysis of collagens solubilized from human foetal, and normal and osteoarthritic adult articular cartilage, with emphasis on type VI collagen. Biochim Biophys Acta. 1990 Apr 19;1038(2):222–230. doi: 10.1016/0167-4838(90)90209-x. [DOI] [PubMed] [Google Scholar]
  33. Ruoslahti E. Fibronectin and its receptors. Annu Rev Biochem. 1988;57:375–413. doi: 10.1146/annurev.bi.57.070188.002111. [DOI] [PubMed] [Google Scholar]
  34. Salter D. M., Godolphin J. L., Gourlay M. S. Chondrocyte heterogeneity: immunohistologically defined variation of integrin expression at different sites in human fetal knees. J Histochem Cytochem. 1995 Apr;43(4):447–457. doi: 10.1177/43.4.7897185. [DOI] [PubMed] [Google Scholar]
  35. Salter D. M., Hughes D. E., Simpson R., Gardner D. L. Integrin expression by human articular chondrocytes. Br J Rheumatol. 1992 Apr;31(4):231–234. doi: 10.1093/rheumatology/31.4.231. [DOI] [PubMed] [Google Scholar]
  36. Schmidt G., Hausser H., Kresse H. Interaction of the small proteoglycan decorin with fibronectin. Involvement of the sequence NKISK of the core protein. Biochem J. 1991 Dec 1;280(Pt 2):411–414. doi: 10.1042/bj2800411. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Shakibaei M., Abou-Rebyeh H., Merker H. J. Integrins in ageing cartilage tissue in vitro. Histol Histopathol. 1993 Oct;8(4):715–723. [PubMed] [Google Scholar]
  38. Smith G. N., Jr, Brandt K. D. Hypothesis: can type IX collagen "glue" together intersecting type II fibers in articular cartilage matrix? A proposed mechanism. J Rheumatol. 1992 Jan;19(1):14–17. [PubMed] [Google Scholar]
  39. Smith G. N., Jr, Hasty K. A., Brandt K. D. Type XI collagen is associated with the chondrocyte surface in suspension culture. Matrix. 1989 Jun;9(3):186–192. doi: 10.1016/s0934-8832(89)80049-6. [DOI] [PubMed] [Google Scholar]
  40. Specks U., Mayer U., Nischt R., Spissinger T., Mann K., Timpl R., Engel J., Chu M. L. Structure of recombinant N-terminal globule of type VI collagen alpha 3 chain and its binding to heparin and hyaluronan. EMBO J. 1992 Dec;11(12):4281–4290. doi: 10.1002/j.1460-2075.1992.tb05527.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Stallcup W. B., Dahlin K., Healy P. Interaction of the NG2 chondroitin sulfate proteoglycan with type VI collagen. J Cell Biol. 1990 Dec;111(6 Pt 2):3177–3188. doi: 10.1083/jcb.111.6.3177. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Streuli C. H., Schmidhauser C., Kobrin M., Bissell M. J., Derynck R. Extracellular matrix regulates expression of the TGF-beta 1 gene. J Cell Biol. 1993 Jan;120(1):253–260. doi: 10.1083/jcb.120.1.253. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Wang A. Z., Zhang X. R., Wang J. C., Fisher G. W., Diamond H. S. Fibronectin induces protease dependent focal matrix depletion and cell overlap in cultured rheumatoid synoviocytes. J Rheumatol. 1995 May;22(5):817–828. [PubMed] [Google Scholar]
  44. Wolf J., Carsons S. E. Distribution of type VI collagen expression in synovial tissue and cultured synoviocytes: relation to fibronectin expression. Ann Rheum Dis. 1991 Jul;50(7):493–496. doi: 10.1136/ard.50.7.493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Yamaguchi Y., Mann D. M., Ruoslahti E. Negative regulation of transforming growth factor-beta by the proteoglycan decorin. Nature. 1990 Jul 19;346(6281):281–284. doi: 10.1038/346281a0. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Anatomy are provided here courtesy of Anatomical Society of Great Britain and Ireland

RESOURCES