Abstract
We undertook a comparative immunolocalisation study on type II collagen, aggrecan and perlecan in a number of 12- to 14-week-old human foetal and postnatal (7–19 months) ovine joints including finger, toe, knee, elbow, hip and shoulder. This demonstrated that perlecan followed a virtually identical immunolocalisation pattern to that of type II collagen in the foetal tissues, but a slightly divergent localisation pattern in adult tissues. Aggrecan was also localised in the cartilaginous joint tissues, which were clearly delineated by toluidine blue staining and the type II collagen immunolocalisations. It was also present in the capsular joint tissues and in ligaments and tendons in the joint, which stained poorly or not at all with toluidine blue. In higher power microscopic views, antibodies to perlecan also stained small blood vessels in the synovial lining tissues of the joint capsule; however, this was not discernable in low power macroscopic views where the immunolocalisation of perlecan to pericellular regions of cells within the cartilaginous rudiments was a predominant feature. Perlecan was also evident in small blood vessels in stromal connective tissues associated with the cartilage rudiments and with occasional nerves in the vicinity of the joint tissues. Perlecan was expressed by rounded cells in the enthesis attachment points of tendons to bone and in rounded cells in the inner third of the meniscus, which stained prominently with type II collagen and aggrecan identifying the chondrogenic background of these cells and local compressive loads. Flattened cells within the tendon and in the surface laminas of articular cartilages and the meniscus did not express perlecan. Collected evidence presented herein, therefore, indicates that besides being a basement membrane component, perlecan is also a marker of chondrogenic cells in prenatal cartilages. In postnatal cartilages, perlecan displayed a pericellular localisation pattern rather than the territorial or interterritorial localisation it displayed in foetal cartilages. This may reflect processing of extracellular perlecan presumably as a consequence of intrinsic biomechanical loading on these tissues or to divergent functions for perlecan and type II collagen in adult compared to prenatal tissues.
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References
Arikawa-Hirasawa E, Watanabe H, Takami H, Hassell JR, Yamada Y (1999) Perlecan is essential for cartilage and cephalic development. Nat Genet 23:354–358
Arikawa-Hirasawa E, Wilcox WR, Le AH, Silverman N, Govindraj P, Hassell JR, Yamada Y (2001a) Dyssegmental dysplasia, Silverman-Handmaker type, is caused by functional null mutations of the perlecan gene. Nat Genet 27:431–434
Arikawa-Hirasawa E, Wilcox WR, Yamada Y (2001b) Dyssegmental dysplasia, Silverman-Handmaker type: unexpected role of perlecan in cartilage development. Am J Med Genet 106:254–257
Arikawa-Hirasawa E, Le AH, Nishino I, Nonaka I, Ho NC, Francomano CA, Govindraj P, Hassell JR, Devaney JM, Spranger J, Stevenson RE, Iannaccone S, Dalakas MC, Yamada Y (2002) Structural and functional mutations of the perlecan gene cause Schwartz-Jampel syndrome, with myotonic myopathy and chondrodysplasia. Am J Hum Genet 70:1368–1375
Benjamin M, Ralphs JR (1998) Fibrocartilage in tendons and ligaments—an adaptation to compressive load. J Anat 193(Pt 4):481–494
Benjamin M, Ralphs JR (2001) Entheses—the bony attachments of tendons and ligaments. Ital J Anat Embryol 106:151–157
Burgeson RE, Hollister DW (1979) Collagen heterogeneity in human cartilage: identification of several new collagen chains. Biochem Biophys Res Commun 87:1124–1131
Costell M, Gustafsson E, Aszodi A, Morgelin M, Bloch W, Hunziker E, Addicks K, Timpl R, Fassler R (1999) Perlecan maintains the integrity of cartilage and some basement membranes. J Cell Biol 147:1109–1122
Edwards AO (2008) Clinical features of the congenital vitreoretinopathies. Eye (Lond) 22:1233–1242
Eyre D (2002) Collagen of articular cartilage. Arthritis Res 4:30–35
Eyre DR, Wu JJ, Fernandes RJ, Pietka TA, Weis MA (2002) Recent developments in cartilage research: matrix biology of the collagen II/IX/XI heterofibril network. Biochem Soc Trans 30:893–899
Eyre DR, Weis MA, Wu JJ (2006) Articular cartilage collagen: an irreplaceable framework? Eur Cell Mater 12:57–63
French MM, Smith SE, Akanbi K, Sanford T, Hecht J, Farach-Carson MC, Carson DD (1999) Expression of the heparan sulfate proteoglycan, perlecan, during mouse embryogenesis and perlecan chondrogenic activity in vitro. J Cell Biol 145:1103–1115
French MM, Gomes RR Jr, Timpl R, Hook M, Czymmek K, Farach-Carson MC, Carson DD (2002) Chondrogenic activity of the heparan sulfate proteoglycan perlecan maps to the N-terminal domain I. J Bone Miner Res 17:48–55
Gleghorn JP, Jones AR, Flannery CR, Bonassar LJ (2007) Boundary mode frictional properties of engineered cartilaginous tissues. Eur Cell Mater 14:20–28 (discussion 28–29)
Gomes RR Jr, Farach Carson MC, Carson DD (2003) Perlecan-stimulated nodules undergo chondrogenic maturation in response to rhBMP-2 treatment in vitro. Connect Tissue Res 44(Suppl 1):196–201
Gustafsson E, Aszodi A, Ortega N, Hunziker EB, Denker HW, Werb Z, Fassler R (2003) Role of collagen type II and perlecan in skeletal development. Ann N Y Acad Sci 995:140–150
Hassell J, Yamada Y, Arikawa-Hirasawa E (2002) Role of perlecan in skeletal development and diseases. Glycoconj J 19:263–267
Iozzo RV (1994) Perlecan: a gem of a proteoglycan. Matrix Biol 14:203–208
Iozzo RV (2005) Basement membrane proteoglycans: from cellar to ceiling. Nat Rev Mol Cell Biol 6:646–656
Iozzo RV, Cohen IR, Grassel S, Murdoch AD (1994) The biology of perlecan: the multifaceted heparan sulphate proteoglycan of basement membranes and pericellular matrices. Biochem J 302(Pt 3):625–639
Jha AK, Yang W, Kirn-Safran CB, Farach-Carson MC, Jia X (2009) Perlecan domain I-conjugated, hyaluronic acid-based hydrogel particles for enhanced chondrogenic differentiation via BMP-2 release. Biomaterials 30:6964–6975
Knox SM, Whitelock JM (2006) Perlecan: how does one molecule do so many things? Cell Mol Life Sci 63:2435–2445
Knox S, Melrose J, Whitelock J (2001) Electrophoretic, biosensor, and bioactivity analyses of perlecans of different cellular origins. Proteomics 1:1534–1541
Knox S, Merry C, Stringer S, Melrose J, Whitelock J (2002) Not all perlecans are created equal: interactions with fibroblast growth factor (FGF) 2 and FGF receptors. J Biol Chem 277:14657–14665
Melrose J, Ghosh P, Taylor TK (2001) A comparative analysis of the differential spatial and temporal distributions of the large (aggrecan, versican) and small (decorin, biglycan, fibromodulin) proteoglycans of the intervertebral disc. J Anat 198:3–15
Melrose J, Smith S, Knox S, Whitelock J (2002) Perlecan, the multidomain HS-proteoglycan of basement membranes, is a prominent pericellular component of ovine hypertrophic vertebral growth plate and cartilaginous endplate chondrocytes. Histochem Cell Biol 118:269–280
Melrose J, Smith S, Ghosh P, Whitelock J (2003) Perlecan, the multidomain heparan sulfate proteoglycan of basement membranes, is also a prominent component of the cartilaginous primordia in the developing human fetal spine. J Histochem Cytochem 51:1331–1341
Melrose J, Smith S, Ghosh P (2004a) Histological and immunohistological studies on cartilage. In: De Ceuninck MS F, Pastoreau P (eds) Cartilage and osteoarthritis volume 2 Structure and in vivo analysis, vol 2. Humana Press, Totowa, pp 39–63
Melrose J, Smith S, Whitelock J (2004b) Perlecan immunolocalises to perichondral vessels and canals in human foetal cartilagenous promordia in early vascular and matrix remodelling events associated with diarthrodial-joint development. J Histochem Cytochem 52:1405–1413
Melrose J, Smith S, Cake M, Read R, Whitelock J (2005a) Comparative spatial and temporal localisation of perlecan, aggrecan and type I, II and IV collagen in the ovine meniscus: an ageing study. Histochem Cell Biol 124:225–235
Melrose J, Smith S, Cake M, Read R, Whitelock J (2005b) Perlecan displays variable spatial and temporal immunolocalisation patterns in the articular and growth plate cartilages of the ovine stifle joint. Histochem Cell Biol 123:561–571
Melrose J, Roughley P, Knox S, Smith S, Lord M, Whitelock J (2006) The structure, location, and function of perlecan, a prominent pericellular proteoglycan of fetal, postnatal, and mature hyaline cartilages. J Biol Chem 281:36905–36914
Melrose J, Hayes AJ, Whitelock JM, Little CB (2008a) Perlecan, the “jack of all trades” proteoglycan of cartilaginous weight-bearing connective tissues. Bioessays 30:457–469
Melrose J, Smith SM, Smith MM, Little CB (2008b) The use of Histochoice for histological examination of articular and growth plate cartilages, intervertebral disc and meniscus. Biotech Histochem 83:47–53
Nakano T, Dodd CM, Scott PG (1997) Glycosaminoglycans and proteoglycans from different zones of the porcine knee meniscus. J Orthop Res 15:213–220
O’Driscoll SW, Commisso CN, Fitzsimmons JS (1995) Type II collagen quantification in experimental chondrogenesis. Osteoarthritis Cartil 3:197–203
Ponsioen TL, van Luyn MJ, van der Worp RJ, van Meurs JC, Hooymans JM, Los LI (2008) Collagen distribution in the human vitreoretinal interface. Invest Ophthalmol Vis Sci 49:4089–4095
Roediger M, Kruegel J, Miosge N, Gersdorff N (2009) Tissue distribution of perlecan domains III and V during embryonic and fetal human development. Histol Histopathol 24:859–868
Smith S, Whitelock JM, Iozzo RV, Melrose J (2009) Topographical variation in the distributions of versican, aggrecan and perlecan in the foetal human spine reflects their diverse functional roles in spinal development. Htistochem Cell Biol 132:491–503
Sun Y, Berger EJ, Zhao C, An KN, Amadio PC, Jay G (2006) Mapping lubricin in canine musculoskeletal tissues. Connect Tissue Res 47:215–221
SundarRaj N, Fite D, Ledbetter S, Chakravarti S, Hassell JR (1995) Perlecan is a component of cartilage matrix and promotes chondrocyte attachment. J Cell Sci 108(Pt 7):2663–2672
Tesche F, Miosge N (2004) Perlecan in late stages of osteoarthritis of the human knee joint. Osteoarthritis Cartil 12:852–862
Tesche F, Miosge N (2005) New aspects of the pathogenesis of osteoarthritis: the role of fibroblast-like chondrocytes in late stages of the disease. Histol Histopathol 20:329–337
Thomopoulos S, Hattersley G, Rosen V, Mertens M, Galatz L, Williams GR, Soslowsky LJ (2002) The localized expression of extracellular matrix components in healing tendon insertion sites: an in situ hybridization study. J Orthop Res 20:454–463
Vincent T, Saklatvala J (2006) Basic fibroblast growth factor: an extracellular mechanotransducer in articular cartilage? Biochem Soc Trans 34:456–457
Vincent T, Hermansson M, Bolton M, Wait R, Saklatvala J (2002) Basic FGF mediates an immediate response of articular cartilage to mechanical injury. Proc Natl Acad Sci USA 99:8259–8264
Vincent TL, Hermansson MA, Hansen UN, Amis AA, Saklatvala J (2004) Basic fibroblast growth factor mediates transduction of mechanical signals when articular cartilage is loaded. Arthritis Rheum 50:526–533
Vincent TL, McLean CJ, Full LE, Peston D, Saklatvala J (2007) FGF-2 is bound to perlecan in the pericellular matrix of articular cartilage, where it acts as a chondrocyte mechanotransducer. Osteoarthritis Cartil 15:752–763
Watanabe H, Yamada Y (2002) Chondrodysplasia of gene knockout mice for aggrecan and link protein. Glycoconj J 19:269–273
Whitelock JM, Melrose J, Iozzo RV (2008) Diverse cell signaling events modulated by perlecan. Biochemistry 47:11174–11183
Yang W, Gomes RR, Brown AJ, Burdett AR, Alicknavitch M, Farach-Carson MC, Carson DD (2006) Chondrogenic differentiation on perlecan domain I, collagen II, and bone morphogenetic protein-2-based matrices. Tissue Eng 12:2009–2024
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Funded by National Health and Medical Research Council of Australia, Project Grant 352562.
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Smith, S.M., Shu, C. & Melrose, J. Comparative immunolocalisation of perlecan with collagen II and aggrecan in human foetal, newborn and adult ovine joint tissues demonstrates perlecan as an early developmental chondrogenic marker. Histochem Cell Biol 134, 251–263 (2010). https://doi.org/10.1007/s00418-010-0730-x
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DOI: https://doi.org/10.1007/s00418-010-0730-x