Abstract
The large cartilage proteoglycan, aggrecan, was found to vary throughout the ovine physis corresponding to the maturational state of the resident chondrocytes. Two populations of proteoglycan monomer were observed in articular, epiphyseal, and in the resting zone of growth plate cartilage. These proteoglycans contained chondroitin sulfate glycosaminoglycan chains sulfated predominantly in the 4 position along with lesser amounts of chondroitin-6-sulfate and keratan sulfate. In the proliferative zone of the growth plate, chondrocytes synthesize one population of proteoglycan monomer which was significantly larger than monomer populations in articular, epiphyseal, or resting zone and this size increase could be attributed to an increase in its constituent chondroitin sulfate side chains. As these chondrocytes progress through their life cycle they continue to modify the structural characteristics of the aggrecan molecule they synthesize. Thus, in the hypertrophic region of the growth plate, the proteoglycan monomer is larger again than in the proliferative region. Variation in sulfation pattern on aggrecan chondroitin sulfate side chains is also observed in the hypertrophic region with an increasing proportion of unsulfated residues present, which may play a role in the initiation of mineralization. In addition, increasing amounts of the carbohydrate sequence recognized by monoclonal antibody 7-D-4 are observed in the hypertrophic zone.
Similar content being viewed by others
References
Ogden JA, Rosenberg LC (1988) Defining the growth plate. In: Uhthoff HK, Wiley JJ (eds) Behaviour of the growth plate. Raven Press Ltd, New York, p 1
Iannotti JP (1990) Growth plate physiology and pathology. Orthop Clin N Am 21:1–17
Howell DS, Dean DD (1992) The biology, chemistry and biochemistry of the mammalian growth plate. In: Coe FL, Favus MJ (eds) Disorders of bone and mineral metabolism. Raven Press Ltd, New York, p 313
Cancedda R, Cancedda FD, Castagnola P (1995) Chondrocyte differentiation. Int Rev Cytol 159:265–358
Schmid TM, Linsenmayer TF (1985) Immunohistochemical localisation of short chain cartilage collagen (type X) in avian tissues. J Cell Biol 100:598–605
Gibson GJ, Flint MH (1985) Type X collagen synthesis by chick sternal cartilage and its relationship to endochondral development. J Cell Biol 101:277–284
Alini M, Matsui Y, Dodge GR, Poole AR (1992)The extra-cellular matrix of cartilage in the growth plate before and during calcification: changes in composition and degradation of type II collagen
Brown RA, Kayser M, McLaughlin B, Weiss JB (1993) Collagenase and gelatinase production by calcifying growth plate chondrocytes. Exp Cell Res 208:1–9
Wuthier RE (1988) Mechanism of matrix vesicle-mediated mineralisation of cartilage. ISI Atlas Sci Biochem 1:231–241
Bianco P, Fisher LW, Young MF, Termine JD, Robey PG (1990) Expression and localization of the two small proteoglycans biglycan and decorin in developing human skeletal and non-skeletal tissues. J Histochem Cytochem 38:1549–1563
Dziewiatkowski DD, Majznerski LL (1985) Role of proteoglycans in endochondral ossification: inhibition of calcification. Calcif Tissue Int 37:560–564
Cuervo LA, Pita JC, Howell DS (1973) Inhibition of calcium phosphate mineral growth by proteoglycan aggregate fractions in a synthetic lymph. Calcif Tissue Res 13:1–10
Chen CC, Boskey AL, Rosenberg LC (1984) The inhibitory effect of cartilage proteoglycans on hydroxyapatite growth. Calcif Tissue Int 36:285–290
Lohmander S, Herpe A (1975) Proteoglycans of mineralizing rib and epiphyseal cartilage. Biochim Biophys Acta 404:93–109
Franzen A, Heinegard D, Reiland S, Olsson SE (1982) Proteoglycans and calcification of cartilage in the femoral head epiphysis of the immature rat. J Bone Jt Surg 64A:558–566
Poole AR, Pidoux I, Rosenberg LC (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
Poole AR, Reddi AH, Rosenberg LC (1982) Persistence of cartilage proteoglycan and link protein during matrix-induced endochondral bone development: an immunofluorescent study. Dev Biol 89:532–539
Scherft JP, Moskalewski S (1984) The amount of proteoglycan in cartilage matrix and the onset of mineralization. Metab Bone Dis Rel Res 5:195–203
Matsui Y, Alini M, Webber C, Poole AR (1991) Characterization of aggregating proteoglycans from the proliferative, maturing, hypertrophic, and calcifying zones of cartilaginous physis. J Bone Jt Surg 73A: 1064–1074
Shepard N, Mitchell N (1985) Ultrastructural modifications of proteoglycans coincident with mineralization in local regions of rat growth plate. J Bone Jt Surg 67A:455–464
Heinegard D, Paulsson M (1984) Structure and metabolism of proteoglycans. In: Piez KA, Reddi AH (eds) Extracellular matrix biochemistry. Elsevier, New York, p 277
Carney SL, Bayliss MT, Collier JM, Muir H (1986) Electrophoresis of 35S-labelled proteoglycans on polyacrylamideagarose composite gels and their visualisation by fluorography. Anal Biochem 156:38–44
Caterson B, Calabro T, Hampton A (1987) Monoclonal antibodies as probes for elucidating proteoglycan structure and function. In: Wight TN, Mecham RP (eds) Biology of proteoglycans. Academic Press, San Diego, p 1
Caterson B, Griffin J, Mahmoodian F, Sorrell JM (1990) Monoclonal antibodies against chondroitin sulphate isomers: their use as probes for investigating proteoglycan metabolism. Biochem Soc Trans 18:820–823
Byers S, Caterson B, Hopwood JJ, Foster BK (1992) Immunolocation analysis of glycosaminoglycans in the human growth plate. J Histochem Cytochem 40:275–282
Sorrell JM, Carrino DA, Caplan AI (1993) Structural domains in chondroitin sulfate identified by anti-chondroitin sulfate monoclonal antibodies. Immunosequencing of chondroitin sulfates. Matrix 13:351–361
Kimura JH, Shinomura T, Thonar EJ-MA (1987) Biosynthesis of cartilage proteoglycan and link protein. Methods Enzymol 144:372–393
Carney SL (1986) Proteoglycans. In: Chaplin MF, Kennedy JF (eds) Carbohydrate analysis: a practical approach. IRL Press, Washington DC, p 97
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Guo YC, Conrad HE (1988) Analysis of oligosaccharides from heparin by reversed-phase ion-pairing high-performance liquid chromatography. Anal Biochem 168:54–62
Blumenkrantz N, Asboe-Hansen G (1973) New method for quantitative determination of uronic acids. Anal Biochem 54: 484–489
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
Bayliss MT, Venn M, Maroudas, A, Ali SY (1983) Structure of proteoglycans from different layers of human articular cartilage. Biochem J 209:387–400
Heinegard D, Wieslander J, Sheehan J, Paulsson M, Sommarin Y (1985) Separation and characterization of two populations of aggregating proteoglycans from cartilage. Biochem J 225:95–106
Caterson B, Mahmoodian F, Sorrell JM, Hardingham TE, Bayliss MT, Carney SL, Ratcliffe A, Muir H (1990) Modulation of native chondroitin sulphate structure in tissue development and in disease. J Cell Sci 97:411–417
Bayliss MT, Ali SY (1978) Age-related changes in the composition and structure of human articular-cartilage proteoglycans. Biochem J 176:683–693
Buckwalter JA, Rosenberg LC, Ungar R (1987) Changes in proteoglycan aggregates during cartilage mineralization. Calcif Tissue Int 41:228–236
Campo RD, Romano JE (1986) Changes in cartilage proteoglycans associated with calcification. Calcif Tissue Int 39: 175–184
Axelsson I, Berman I, Pita JC (1983) Proteoglycans from rabbit articular and growth plate cartilage. Ultracentrifugation, gel chromatography, and electron microscopy. J Biol Chem 258:8915–8921
Plaas AHK, Sandy JD (1993) A cartilage explant system for studies on aggrecan structure, biosynthesis and catabolism in discrete zones of the mammalian growth plate. Matrix 13: 135–147
Kimata K, Okayama M, Ooira A, Suzuki S (1974) Heterogeneity of proteochondroitin sulfates produced by chondrocytes at different stages of cytodifferentiation. J Biol Chem 249: 1646–1653
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
Stocum DL, Davis RM, Leger M, Conrad HE (1979) Development of the tibiotarsus in the chick embryo: biosynthetic activities of histologically distinct regions. J Embryol Exp Morph 54:155–170
Farquharson C, Whitehead CC, Loveridge N (1994) Alterations in glycosaminoglycan concentration and sulfation during chondrocyte maturation. Calcif Tissue Int 54:296–303
Matthews MB (1964) Structural factors in cation binding in anionic polysaccharides of connective tissue. Arch Biochem Biophys 104:394–404
Hunter GK, Wong KS, Kim JJ (1988) Binding of calcium to glycosaminoglycans: an equilibrium dialysis study. Arch Biochem Biophys 260:161–167
Althoff J, Quint P, Krefting ER, Hohling HJ (1982) Morphological studies on the epiphyseal growth plate combined with biochemical and X-ray microprobe analyses. Histochemistry 74:541–552
Shapiro IM, Boyde A (1984) Microdissection: elemental analysis of the mineralizing growth cartilage of the normal and rachitic chick. Metab Bone Dis Rel Res 5:317–326
Shaklee PN, Conrad HE (1985) Structural changes in the large proteoglycan in differentiating chondrocytes from the chick embryo tibiotarsus. J Biol Chem 260:16064–16067
Sorrell JM, Lintala AM, Mahmoodian F, Caterson B (1988) Epitope-specific changes in chondroitin sulfate/dermatan sulfate proteoglycans as markers in the lymphopoietic and granulopoietic compartments of developing bursae of fabricius. J Immunol 140:4263–4270
Sorrell JM, Mahmoodian F, Schafer IA, Davis B, Caterson B (1990) Identification of monoclonal antibodies that recognized novel epitopes in native chondroitin/dermatan glycosaminoglycan chains: their use in mapping functionally distinct domains of human skin. J Histochem Cytochem 38:393–402
Carney SL, Billingham ME, Caterson B, Ratcliffe A, Bayliss MT, Hardingham TE, Muir H (1992) Changes in proteoglycan turnover in experimental canine osteoarthritic cartilage. Matrix 12:137–147
Turnbull JE, Fernig DG, Ke Y, Wilkinson MC, Gallagher JT (1992) Identification of the basic fibroblast growth factor binding sequence in fibroblast heparan sulfate. J Biol Chem 267:10337–10341
Lindahl U, Thunberg L, Backstrom G, Riesenfeld J, Nordling K, Bjork I (1984) Extension and structural variability of the antithrombin-binding sequence in heparin. J Biol Chem 259: 12368–12376
Maimone MM, Tollefsen DM (1990) Structure of a dermatan sulfate hexasaccharide that binds to heparin cofactor II with high affinity. J Biol Chem 265:18263–18271
Hardingham TE, Fosang AJ (1992) Proteoglycans: many forms and many functions. FASEB 6:861–870
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Byers, S., van Rooden, J.C. & Foster, B.K. Structural changes in the large proteoglycan, aggrecan, in different zones of the ovine growth plate. Calcif Tissue Int 60, 71–78 (1997). https://doi.org/10.1007/s002239900188
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s002239900188