Skip to main content
Log in

Quantitative immunohistochemical evidence of a functional gradient of chondroitin 4-sulphate/dermatan sulphate, developmentally regulated in the predentine of rat incisor

  • Published:
The Histochemical Journal Aims and scope Submit manuscript

Abstract

A quantitative examination was carried out on the early and mature stages of dentinogenesis in the rat incisor, using a post-embedding immunogold labelling with an anti-chondroitin 4 sulphate/dermatan sulphate antibody (2B6). At a very early stage of predentine formation, before polarizing odontoblasts have established junctional complexes, immunolabelling was weak. In contrast, when polarized odontoblasts established distal junctional complexes, immunolabelling in predentine was uniform and threefold denser than in initial predentine. The same gold particle density was found in the non-mineralized mantle dentine. During circumpulpal dentine formation, a gradient was seen in predentine, a larger number of gold particles being scored in the proximal zone compared with the distal region adjacent to the mineralization front. In circumpulpal dentine, some labelling was found within the lumen of the tubules and in the bordering dentine around the tubules. A few particles were also detected in intertubular matrix after demineralization. Together, these data provide evidence for a developmentally regulated gradient during the transition between mantle and circumpulpal dentine, and also in a more mature part of the tooth, a functional gradient that probably plays a role in the process of mineralization. © 1998 Chapman & Hall

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Blumenthal, N.C., Posner, A.S., Silverman, L.D. & Rosenberg, L.C. (1979) Effect of proteoglycans on in vitro hydroxyapatite formation. Calcif. Tissue Int. 27, 75–82.

    Google Scholar 

  • Branford white, C.J. (1978) Molecular organization of heparan sulphate proteoglycan from human dentine. Arch. Oral Biol. 23, 1141–44.

    Google Scholar 

  • Byers, S., Caterson, B., Hopwood, J. J. & Foster, B.K. (1992) Immunolocation analysis of glycosaminoglycans in the human growth plate. J. Histochem. Cytochem. 40, 275–82.

    Google Scholar 

  • Caterson, B., Christner, J.E., Baker, J.R. & Couchman, J.R. (1985) Production and characterization of monoclonal antibodies directed against connective tissue proteoglycans. Fed. Proc. 44, 386–93.

    Google Scholar 

  • Chen, C.-C. & Boskey, A.L. (1985) Mechanisms of proteoglycan inhibition of hydroxyapatite growth. Calcif. Tissue Int. 37, 395–400.

    Google Scholar 

  • Chen, C.-C., Boskey, A.L. & Rosenberg, L.C. (1984) The inhibitory effect of cartilage proteoglycans on hydroxyapatite growth. Calcif. Tissue Int. 36, 285–90.

    Google Scholar 

  • Clark, R.D., Smith, J.G. & Davidson, E.A. (1965) Hexoamine and acid glycosaminoglycans in human teeth. Biochim. Biophys. Acta 101, 267–72.

    Google Scholar 

  • Desjardins, M. & Bendayan, M. (1989) Heterogenous distribution of type IV collagen, entactin, heparan sulfate proteoglycan, and laminin among renal basement membranes as demonstrated by quantitative immunocytochemistry. J. Histochem. Cytochem. 37, 885–97.

    Google Scholar 

  • Embery, G. (1974) The isolation of chondroitin 4-[35S] sulphate from the molar teeth of young rats receiving sodium [35S] sulphate. Calcif. Tissue Res. 14, 59–65.

    Google Scholar 

  • Embery, G. & Smalley, J.W. (1980) The influence of fluoride on the uptake of radiosulphate by rat incisor odontoblasts in vitro. Arch. Oral Biol. 25, 659–62.

    Google Scholar 

  • Fukae, M., Tanabe, T. & Yamada, M. (1994) Action of metalloproteinases on porcine dentine mineralization. Calcif. Tissue Int. 55, 426–35.

    Google Scholar 

  • Galbraith, D.B., Cutler, L.S. & Kollar, E.J. (1992) The correlation of temporal regulation of glycosaminoglycan synthesis with morphogenetic events in mouse tooth development. Arch. Oral Biol. 37, 623–8.

    Google Scholar 

  • Goldberg, H.A., Domenicucci, C., Pringle, G.A. & Sodek, J. (1988) Mineral-binding proteoglycans of fetal porcine calvarial bone. J. Biol. Chem. 263, 12092–101.

    Google Scholar 

  • Goldberg, M. & Escaig, F. (1984) The appearance in TEM of proteoglycan predentine is fixation dependant. J. Microscopy 134, 161–7.

    Google Scholar 

  • Goldberg, M. & Escaig, F. (1985) Incorporation of (35S) sulfate and (3H) glucosamine into glycosaminoglycans in rat incisor predentine and dentine: comparison by autoradiography of fixation by rapid-freezing, freeze-substitution, and aldehyde fixation. Calcif. Tissue Int. 37, 511–18.

    Google Scholar 

  • Goldberg, M. & Septier, D. (1983) Electron microscopic visualization of proteoglycansin rat incisor predentine and dentine with cuprolinic blue. Arch. Oral Biol. 28, 79–83.

    Google Scholar 

  • Goldberg, M. & Septier, D. (1986) Visualization of proteoglycans and membrane associated components in rat incisor predentine and dentine using ruthenium hexamine trichloride. Arch. Oral Biol. 31, 205–12.

    Google Scholar 

  • Goldberg, M. & Septier, D. (1996) A comparative study of the transition between predentin and dentin, using various preparative procedures in the rat. Eur. J. Oral Sci. 104, 269–77.

    Google Scholar 

  • Goldberg, M. & Takagi, M. (1993) Dentine proteoglycans: composition, ultrastructure and functions. Histochem. J. 25, 781–806.

    Google Scholar 

  • Goldberg, M., Septier, D. & Escaig-Haye, F. (1987) Glycoconjugates in dentinogenesis and dentin. Prog. Histochem. Cytochem. 17, 1–113.

    Google Scholar 

  • Hall, R.C., Embery, G. & Shellis, R.P. (1996a) Fluoride modulates the inhibition of in vitro hydroxyapatite crystal growth by small proteoglycans: relevance to dental calculus. Adv. Dent. Res. 9, 433–42.

    Google Scholar 

  • Hall, R.C., Embery, G. & Waddington, R.J. (1996b) The structure of the proteoglycans of rat incisor dentinepredentine is modified during in vivo induced fluorosis. Eur. J. Oral Sci. 104, 285–91.

    Google Scholar 

  • Hall, R.C., Lloyd, D. & Embery, G. (1997) The immunolocalisation of lumican in human predentine. Arch. Oral Biol. 42, 783–86.

    Google Scholar 

  • Herjpe, A. & Engfeldt, B. (1976) Proteoglycans of dentine and predentine. Calcif. Tissue Res. 22, 173–82.

    Google Scholar 

  • Herjpe, A., Antonopoulos, C.A., Engfeldt, B. & WikstrÖm, B. (1983) Analysis of dentine glycosaminoglycans using high-performance liquid chromatography. Calcif. Tissue Int. 35, 496–501.

    Google Scholar 

  • Hunter, G.K. (1991) Role of proteoglycan in the provisional calcification of cartilage. A review and reinterpretation. Clin. Orthop. 262, 256–80.

    Google Scholar 

  • Iozzo, R.V. & Murdoch, A.D. (1996) Proteoglycans of the extracellular environment: clues from the gene and protein side offer novel perspectives in molecular diversity and function. FASEB J. 10, 598–614.

    Google Scholar 

  • Jones, I.L. & Leaver, A.G. (1974) Glycosaminoglycans of human dentine. Calcif. Tissue Res. 16, 37–44.

    Google Scholar 

  • Kresse, H., Hausser, H. & Schö nherr, E. (1993) Small proteoglycans. Experientia 49, 405–16.

    Google Scholar 

  • Leblond, C.P. & Inoue, S. (1989) Structure, composition, and assembly of basement membrane. Am. J. Anat. 185, 367–90.

    Google Scholar 

  • Lin, X.-H., Dahlin-Huppe, K. & Stallcup, W.B. (1996) Interaction of the NG2 proteoglycan with actin cytoskeleton. J. Cell. Biochem. 63, 463–77.

    Google Scholar 

  • Linde, A., Bhown, M. & Butler, W.T. (1980) Noncollagenous proteins of dentin. A re-examination of proteins from rat incisor dentin utilizing techniques to avoid artifacts. J. Biol. Chem. 255, 5931–42.

    Google Scholar 

  • Linde, A., Lussi, A. & Crenshaw, M.A. (1989) Mineral induction by immobilized proteins. Calcif. Tissue Int. 44, 286–95.

    Google Scholar 

  • Lormeé e, P., Septier, D., Leécolle, S., Baudoin, C. & Goldberg, M. (1996) Dual incorporation of (35S) sulfate into dentin proteoglycans acting as mineralization promoters in rat molars and predentin proteoglycans. Calcif. Tissue Int. 58, 368–75.

    Google Scholar 

  • Mark, M.P., Baker, J.R., Kimata, K., Ruch, J.-V. (1990a) Regulated changes in chondroitin sulfation during embryogenesis: an immunohistochemical approach. Int. J. Dev. Biol. 34, 191–204.

    Google Scholar 

  • Mark, M.P., Baker, J.R., Morrison, K. & Ruch, J.-V. (1990b) Chondroitin sulfates in developing tooth germs. An immunohistochemical study with monoclonal antibodies against chondroitin-4 and chondroitin-6 sulfates. Differentiation 43, 37–50.

    Google Scholar 

  • Mark, M.P., Karcher-Djuricic, V., Baker, J.R. & Ruch, J.-V. (1990c) Effects of â-D-xyloside on morphogenesis and cytodifferentiation in cultured embryonic mouse molars. Cell Differ. Dev. 32, 1–16.

    Google Scholar 

  • Nader, H.B., Oliveira, F.W., Jeronimo, S.M.B., Chavante, S.F., Sampaio, L.O. & Dietrich, C.P. (1996) Synchronized order of appearance of hyaluronic acid (or acidic galactan) ← chondroitin C-6 sulfate ←chondroitin C-4/C-6 sulfate, heparan sulfate, dermatan sulfate ← heparin during morphogenesis, differentiation and development. Braz. J. Med. Biol. Res. 29, 1221–6.

    Google Scholar 

  • Poole, A.R., 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, 249–60.

    Google Scholar 

  • Rahemtulla, F., Prince, C.W. & Butler, W.T. (1984) Isolation and partial characterization of proteoglycans from rat incisors. Biochem. J. 218, 877–85.

    Google Scholar 

  • SchÖnherr, E., Witsch-Prehm, P., Harrach, B., Robenek, H., Rauterberg, J. & Kresse, H. (1995) Interaction of biglycan with type I collagen. J. Biol. Chem. 270, 2776–83.

    Google Scholar 

  • Scott, J.E. (1988) Proteoglycan-fibrillar collagen interactions. Biochem. J. 252, 313–23.

    Google Scholar 

  • Steinfort, J., Van de stadt, R. & Beertsen, W. (1994) Identification of new rat dentin proteoglycans utilizing C18 chromatography. J. Biol. Chem. 269, 22397–404.

    Google Scholar 

  • SundstrÖm, B. (1971) New aspects on the utilization of inorganic sulphate during dentin formation. Histochemie 26, 61–6.

    Google Scholar 

  • Takagi, M., Parmley, R.T. & Denys, F.R. (1981) Ultrastructural localization of complex carbohydrates in odontoblasts, predentin, and dentin. J. Histochem. Cytochem. 29, 747–58.

    Google Scholar 

  • Takagi, M., Hishikawa, H., Hosokawa, Y., Kagami, A. & Rahemtulla, F. (1990) Immunohistochemical localization of glycosaminoglycans and proteoglycans in predentin and dentin of rat incisors. J. Histochem. Cytochem. 38, 319–24.

    Google Scholar 

  • Timpl, R. (1993) Proteoglycans of basement membranes. Experientia 49, 417–28.

    Google Scholar 

  • Warshawsky, H. & Moore, G. (1976) A technique for the fixation and decalcification of rat incisors for electron microscopy. J. Histochem. Cytochem. 15, 542–9.

    Google Scholar 

  • Weinstock, A. (1972) Matrix development in mineralizing tissues as shown by radioautography formation of enamel and dentin. In Developmental Aspects of Oral Biology (edited by Slavkin, H.C. & Bavetta, L.A.) pp. 201–42. New York: Academic Press.

    Google Scholar 

  • Yoshiba, N., Yoshiba, K., Iwaku, M. & Ozawa, H. (1996) Immunolocalization of the small proteoglycan decorin in human teeth. Arch. Oral Biol. 41, 351–7.

    Google Scholar 

  • Yurchenco, P.D. & Schittny, J.C. (1990) Molecular architecture of basement membranes. FASEB J. 4, 1577–90.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

SEPTIER, D., HALL, R.C., LLOYD, D. et al. Quantitative immunohistochemical evidence of a functional gradient of chondroitin 4-sulphate/dermatan sulphate, developmentally regulated in the predentine of rat incisor. Histochem J 30, 275–284 (1998). https://doi.org/10.1023/A:1003216024158

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1003216024158

Keywords

Navigation