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Alterations in glycosaminoglycan concentration and sulfation during chondrocyte maturation

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Abstract

We have used antibodies to chondroitin 4- and 6-sulfate and keratan sulfate along with Alcian blue staining of sulfated proteoglycans to investigate changes in content and sulfation within the avian growth plate. In normal chicks, chondroitin 4- and 6-sulfate content were similar in the proliferating and transitional zones but in the hypertrophic zone, chondroitin 4- and 6-sulfate were slightly lower (13% and 18%, respectively) and keratan sulfate was markedly lower (58%). Compared with the proliferative zone, Alcian blue staining of sulfated glycosaminoglycans was markedly lower in both the transitional (46%) and hypertrophic (22%) zones. In tibial dyschondroplasia, where chondrocyte maturation is arrested at the transitional zone, there was no difference in the chondroitin 4- and 6-sulfate or keratan sulfate staining between the proliferative and transitional zones, which were similar to normal birds. With Alcian blue staining there was no difference in the intensity of the staining within the proliferating zone compared with normal birds but stainign in the transitional chondrocytes was markedly higher (39%). These results suggest that in the early steps of chondrocyte maturation there may be a decrease in the degree of glycosaminoglycan sulfation without any alteration in glycosaminoglycan concentration, and that further maturation may be accompanied by a change in the nature of the proteoglycans which may also affect the level of sulfation.

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References

  1. Isaksson OGP, Lindahl A, Nilsson A, Isgaard J (1987) Mechanism of the stimulatory effect of growth hormone on longitudinal bone growth. Endocr Rev 8:426–438

    Google Scholar 

  2. Loveridge N, Thomson BM, Farquharson C (1992) Bone growth and turnover. In: Whitehead CC (ed) Bone biology and skeletal disorders of poultry. Carfax Publishing Co, Oxford, p 3

    Google Scholar 

  3. Solursh M, Jensen KL, Reiter RS, Schmid TM, Linsenmayer TF (1984) Extracellular-matrix mediates epithelial effects of chondrogenesis in vitro. Dev Biol 105:451–457

    Google Scholar 

  4. Sommarin Y, Larsson T, Heinegard D (1989) Chondrocytematrix interactions. Exp Cell Res 184:181–192

    Google Scholar 

  5. Kwan APL, Freemont AJ, Grant ME (1986) Immunoperoxidase localisation of type X collagen in chick tibiae. Biosci Rep 6:155–162

    Google Scholar 

  6. Appleton J (1988) The ultrastructural distribution of proteoglycans in normal and rachitic growth cartilage from the mandibular condyle of the rat. Arch Oral Biol 33:379–381

    Google Scholar 

  7. Leach RM, Nesheim MC (1965) Nutritional, genetic and morphological studies of an abnormal cartilage formation in young chicks. J Nutr 86:236–244

    Google Scholar 

  8. Riddell C (1971) Studies on the pathogenesis of tibial dyschondroplasia in chickens. 1. Production of a similar defect by surgical interference. Avian Dis 15:557–565

    Google Scholar 

  9. Farquharson C, Whitehead CC, Rennie S, Thorp BH, Loveridge N (1992) Cell proliferation and enzyme activities associated with the development of avian tibial dyschondroplasia: an in situ biochemical study. Bone 13:59–67

    Google Scholar 

  10. Loveridge N, Farquharson C, Hesketh JE, Jakowlew SB, Whitehead CC, Thorp BH (1993) The control of chondrocyte differentiation during endochondral bone growth in vivo: changes in TGF-β and the proto-oncogene c-myc. J Cell Sci 105:949–956

    Google Scholar 

  11. Buckwalter JA, Rosenberg LC, Ungar, R (1987) Changes in proteoglycan aggregates during cartilage mineralization. Calcif Tissue Int 41:228–236

    Google Scholar 

  12. Campo RD, Romano JE (1986) Changes in cartilage proteoglycans associated with calcification. Calcif Tissue Int 39:175–184

    Google Scholar 

  13. Buckwalter JA, Rosenberg L (1986) Structural changes in reassembled growth cartilage aggregates. J Orthop Res 4:1–9

    Google Scholar 

  14. Horton WA, Machado MM (1988) Extracellular matrix alterations during endochondral ossification in humans. J Orthop Res 6:793–803

    Google Scholar 

  15. Muir H (1983) Proteoglycans as organisers of the intercellular matrix. Biochem Soc Trans 11:613–622

    Google Scholar 

  16. Heinegard D, Sommarin Y (1987) Proteoglycans: an overview. Methods Enzymol 144:305–319

    Google Scholar 

  17. Hardingham TE, Fosang AJ (1992) Proteoglycans: many forms and many functions. FASEB J 6:861–870

    Google Scholar 

  18. Oldberg A, Antonsson P, Hedbom E, Heinegard D (1990) Structure and function of extracellular matrix proteoglycans. Biochem Soc Trans 18:789–792

    Google Scholar 

  19. Pita JC, Cuervo LA, Madruga JE, Muller FJ, Howell DS (1970) Evidence for a role of protein polysaccharide association with the onset of calcification in cartilage. J Clin Invest 49:2188–2196

    Google Scholar 

  20. de Bernard B, Stagni N, Colautti I, Vittur F, Bonucci E (1977) Glycosaminoglycans and endochondral calcification. Clin Orthop Rel Res 126:285–291

    Google Scholar 

  21. Reddi AH, Kuettner KE (1981) Vascular invasion of cartilage: correlation of morphology with lysozyme, glycosaminoglycans, protease, and protease inhibitory activity during endochondral bone development. Dev Biol 82:217–223

    Google Scholar 

  22. Futami T, Ototani N, Nagatsuka Y, Yosizawa Z (1979) Comparison of carbohydrate-containing substances from noncalcified and calcified portions of bovine costal cartilage. J Biochem 85:1067–1073

    Google Scholar 

  23. Boyd A, Shapiro IM (1980) Energy dispersive x-ray elemental analysis of isolated epiphysael growth plate chondrocyte fragments. Histochemistry 69:85–94

    Google Scholar 

  24. Orkin RW, Williams BR, Cranley RE, Poppke DC, Brown KS (1977) Defects in the cartilaginous growth plates of brachymorphic mice. J Cell Biol 73:287–299

    Google Scholar 

  25. Boskey AL, Maresca M, Wilkstrom B, Hjerpe A (1991) Hydroxyapatite formation in the presence of proteoglycans of reduced sulfate content: studies in the brachymorphic mouse. Calcif Tissue Int 49:389–393

    Google Scholar 

  26. Luft JH (1971) Ruthenium red and violet II. Fine structural localization in animal tissues. Anat Rec 171:369–377

    Google Scholar 

  27. Shepard NL, Mitchell S (1976) The localisation of proteoglycan by light and electron microscopy using safranin O. J Ultrastruct Res 54:451–460

    Google Scholar 

  28. Schofield BH, Williams BR, Doty SB (1975) Alcian blue staining of cartilage for electron microscopy. Application of the critical electrolyte concentration principle. Histochem J 7:139–145

    Google Scholar 

  29. Couchman JR, Caterson B, Christner JE, Baker JR (1984) Mapping monoclonal antibody detection of glycosaminoglycans in connective tissue. Nature 307:650–652

    Google Scholar 

  30. Caterson B, Christner JE, Baker JR, Couchman JR (1985) Production and characterization of monoclonal antibodies directed against connective tissue proteoglycans. Fed Proc 44: 386–393

    Google Scholar 

  31. Zanetti M, Ratcliffe A, Watt FM (1985) Two subpopulations of the differentiated chondrocytes identified with a monoclonal antibody to keratan sulphate. J Cell Biol 101: 53–59

    Google Scholar 

  32. Mehmet H, Scudder P, Tang PW, Hounsell EF, Caterson B, Feizi T (1986) The antigenic determinants recognised by three monoclonal antibodies to keratan sulphate involve sulphated hepta- or larger oligosaccharides of the poly (N-acetyllactosamine) series. Eur J Biochem 157: 385–391

    Google Scholar 

  33. Thorp BH, Whitehead CC, Rennie JS (1991) Avian tibial dyschondroplasis: a comparison of the incidence and severity as assessed by gross examination and histopathology. Res Vet Sci 51: 48–54

    Google Scholar 

  34. Poole AR, Pidoux I (1989) Immunoelectron microscopic studies of type X collagen in endochondral ossification. J Cell Biol 109: 2547–2554

    Google Scholar 

  35. Sorrell JM, Mahmoodian F, Caterson B (1988) Immunochemical characterization and ultrastructural localization of chondroitin sulfates and keratan sulfate in embryonic chick bone marrow. Cell Tissue Res 252: 523–531

    Google Scholar 

  36. Smith JC, Watt FM (1985) Biochemical specificity of Xenopus notochord. Differentiation 29: 109–115

    Google Scholar 

  37. Scott JE, Dorling J (1965) Differential staining of acidic glycosaminoglycans (mucopolysaccharides) by Alcian blue in salt solutions. Histochemie 5: 221–233

    Google Scholar 

  38. Chayen J, Bitensky L (1991) Practical histochemistry, 2nd ed. John Wiley & Sons, London, New York

    Google Scholar 

  39. Ealey PA, Henderson B, Loveridge N (1984) A quantitative study of peroxidase activity in unfixed tissue of the guinea pig thyroid gland. Histochem J 16: 111–122

    Google Scholar 

  40. Horne Z, Hesketh HE (1990) Immunological localization of ribosomes in striated rat muscle. Biochem J 268: 231–236

    Google Scholar 

  41. Dunham J, Shackleton DR, Nahir AM, Billingham EJ, Bitensky L, Chayen J, Muir H (1985) Altered orientation of glycosaminoglycans and cellular changes in the tibial cartilage in the first two weeks of experimental canine osteoarthritis. J Orthop Res 3: 258–268

    Google Scholar 

  42. Jibril O (1967) Proteolytic degradation of ossifying cartilage matrix and the removal of acid mucopolysaccharides prior to bone formation. Biochim Biophys Acta 136: 162–165

    Google Scholar 

  43. Suzuki K, Takase T, Takigawa M, Uchida A, Shimomur Y (1981) Simulation of the initial stage of endochondral ossification: in vitro sequential culture of growth cartilage cells and bone marrow cells. Proc Natl Acad Sci USA 78: 2368–2372

    Google Scholar 

  44. Dziewiatkowski DD, Majznerski LL (1985) Role of proteoglycans in endochondral ossification. Calcif Tissue Int 37: 560–567

    Google Scholar 

  45. Hunter GK, Grynpas MD, Cheng P-T, Pritzker KPH (1987) Effect of glycosaminoglycans on calcium pyrophosphate crystal formation in collagen gels. Calcif Tissue Int 41: 164–169

    Google Scholar 

  46. Chen CC, Boskey AL, Rosenberg LC (1984) The inhibitory effect of cartilage proteoglycans on hydroxyapatite growth. Calcif Tissue Int 36: 285–290

    Google Scholar 

  47. Bianco P, Fisher LW, Young MF, Termine JD, Robey PG (1990) Expression and localisation of the two small proteoglycans biglycan and decorin in developing human skeletal and non-skeletal tissues. J Histochem Cytochem 38: 1549–1563

    Google Scholar 

  48. Robey PG, Bianco P, Termine JD (1992) The cellular biology and molecular biochemistry of bone formation. In: Coe FL, Favus MJ (eds) Disorders of bone and mineral metabolism. Raven Press Ltd, New York, p 241

    Google Scholar 

  49. Scherft J, Moskalewski S (1984) The amount of proteoglycans in cartilage matrix and the onset of mineralisation. Metab Bone Dis Rel Res 5: 195–203

    Google Scholar 

  50. Matsui Y, Alini M, Webber C, Poole AR (1991) Characterization of aggregating proteoglycans from the proliferative, maturing, hypertrophic and calcifying zones of the cartilaginous physis. J Bone Joint Surg 73-A: 1064–1074

    Google Scholar 

  51. Arsenault AL, Ottensmeyer FP (1983) Quantitative spatial distributions of calcium, phosphorus, and sulfur in calcifying epiphysis by high resolution electron microscopic imaging. Proc Natl Acad Sci USA 80: 322–1326

    Google Scholar 

  52. Hunter GK (1991) Role of proteoglycan in the provisional calcification of cartilage. A review and reinterpretation. Clin Orthop 262: 265–280

    Google Scholar 

  53. Barckhaus RH, Krefting ER, Althoff J, Quint P, Hohling HJ (1971) Electron-microscopic microprobe analysis on the initial stages of mineral formation in the epiphyseal growth plate. Cell Tissue Res 217: 661–670

    Google Scholar 

  54. Weatherell JA, Weidmann SM (1963) The distribution of organically bound sulfate in bone and cartilage calcification. Biochem J 89: 265–273

    Google Scholar 

  55. Sorrell JM, Caterson B (1989) Detection of age-related changes in the distribution of keratan sulfates and chondroitin sulfates in developing chick limbs: an immunocytochemical study. Development 106: 657–663

    Google Scholar 

  56. Byers S, Caterson B, Hopwood JJ, Foster BK (1992) Immunolocation analysis of glycosaminoglycans in the human growth plate. J Histochem Cytochem 40: 275–282

    Google Scholar 

  57. Takagi M, Maeno M, Kagami A, Takahashi Y, Otsuka K (1991) Biochemical and immunocytochemical characterization of mineral binding proteoglycans in rat bone. J Histochem Cytochem 39: 41–50

    Google Scholar 

  58. Poole AR, Pidoux I, Rosenberg L (1982) Role of proteoglycans in endochondral ossification, immunofluorescent localization of link protein and proteoglycan monomer in bovine fetal epiphyseal growth plate. J Cell Biol 92: 249–260

    Google Scholar 

  59. Ecarot-Charrier B, Bouchard F, Delloye C (1989) Bone sialoprotein II synthesised by cultures osteoblasts contains tyrosine sulfate. J Biol Chem 264: 20049–20053

    Google Scholar 

  60. Midura RJ, McQuillan DJ, Benham KJ, Fisher LW, Hascall VC (1990) A rat osteogenic cell line (UMR 106-01) synthesises a highly sulfated form of bone sialoprotein. J Biol Chem 256: 5285–5291

    Google Scholar 

  61. Bianco P, Fisher LW, Young MF, Termine JD, Robey PG (1991) Expression of bone sialoprotein (BSP) in developing human tissues. Calcif Tissue Int 49: 421–426

    Google Scholar 

  62. Reinholt FP, Engfeldt B, Heinegard D, Hjerpe A (1985) Proteoglycans and glycosaminoglycans of normal and strontium rachitic epiphyseal cartilage. Collagen Rel Res 5: 41–53

    Google Scholar 

  63. Orkin RW, Pratt RM, Martin GR (1976) Undersulphated chondroitin sulfate in the cartilage matrix of brachymorphic mice. Dev Biol 50: 82–94

    Google Scholar 

  64. Pennypacker JP, Kimata K, Brown KS (1981) Brachymorphic mice (bm/bm): a generalized biochemical defect expressed primarily in cartilage. Dev Biol 81: 280–287

    Google Scholar 

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Authors and Affiliations

  1. AFRC Roslin Institute (Edinburgh), Roslin, Midlothian, Scotland

    C. Farquharson & C. C. Whitehead

  2. Biochemical Sciences, Rowett Research Institute, Bucksburn, Aberdeen, Scotland

    C. Farquharson & N. Loveridge

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  1. C. Farquharson
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  2. C. C. Whitehead
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  3. N. Loveridge
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Farquharson, C., Whitehead, C.C. & Loveridge, N. Alterations in glycosaminoglycan concentration and sulfation during chondrocyte maturation. Calcif Tissue Int 54, 296–303 (1994). https://doi.org/10.1007/BF00295954

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  • DOI: https://doi.org/10.1007/BF00295954

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