Summary
Decalcified and nondecalcified sections of fetal bovine tibia were stained immunohistochemically with a monoclonal antibody against dentin phosphophoryn. In the epiphyseal portion of the long bone, osteoblasts, osteocytes and the bone matrix were stained, but chondrocytes and the cartilage matrix were not. Similar staining was observed in the epiphyseal and diaphyseal portions of bones. These findings suggest that a protein(s) with the same epitope as phosphophoryn may be synthesized and secreted by osteoblasts at the beginning of ossification and may be involved in mineralization of bone tissue. On Western blots of proteins extracted from fetal bovine bone, the antibody reacted with two bands of molecular weights of about 71,000 and 63,000. These proteins and antibody(s) to the proteins may be useful for detection of the phenotype of osteogenesis
This is a preview of subscription content, log in via an institution to check access.
References
Butler WT (1984) Matrix macromolecules of bone and dentin. Coll Relat Res 4: 297–307
Fisher LW, Termine J (1985) Noncollagenous proteins influencing the local mechanisms of calcification. Clin Orthop Relat Res 200: 362–385
Price PA, Otsuka AS, Poser JW, Kristaponis J, Raman N (1976) Characterization of a γ-car-boxyglutamic acid-containing protein from bone. Proc Natl Acad Sci USA 73: 1447–1451
Termine JD, Kleinman HK, Whitson SW, Conn KM, McGarvey ML, Martin GR (1981) Osteonectin, a bone-specific protein linking mineral to collagen. Cell 26: 99–105
Price PA, Urist MR, Otawara Y (1983) Matrix Gla protein, a new γ-carboxyglutamic acid-containing protein which is associated with the organic matrix of bone. Biochem Biophys Res Commun 117: 765–771
Sato S, Rahemtulla F, Prince CW, Tomana M, Butler WT (1985) Acidic glycoproteins from bovine compact bone. Connect Tissue Res 4: 51–64
Oldberg Å, Franzén A, Heinegård D (1986) Cloning and sequence analysis of rat bone sialoprotein (osteopontin) cDNA reveals an Arg-Gly-Asp cell-binding sequence. Proc Natl Acad Sci USA 83: 8819–8823
Uchiyama A, Suzuki M, Lefteriou B, Glimcher M (1986) Isolation and chemical characterization of the phosphoproteins of chicken bone matrix: heterogeneity in molecular weight and composition. Biochemistry 25: 7572–7583
Mardon HJ, Triffitt JT (1987) A tissue specific protein in rat osteogenic tissue. J Bone Mineral Res 2: 191–199
Fisher LW, Hawkins GR, Tuross N, Termine JD (1987) Purification and partial characterization of small proteoglycans I and II, bone sialoproteins I and II, and osteonectin from the mineral compartment of developing human bone. J Biol Chem 262: 9702–9708
Mark MP, Prince CW, Gay S, Austin RL, Bhown M, Finkelman RD, Butler WT (1987) A comparative immunocytochemical study on the subcellular distributions of 44 kDa bone phosphoprotein and bone γ-carboxyglutamic acid (Gla)-containing protein in osteoblasts. J Bone Mineral Res 2: 337–346
Nakamura O, Gohda E, Ozawa M, Senba I, Miyazaki H, Murakami T, Daikuhara Y (1985) Immunohistochemical studies with a monoclonal antibody on the distribution of phosphophoryn in predentin and dentin. Calcif Tissue Int 37: 491–500
McLean IW, Nakane PK (1974) Periodate-lysine-paraformaldehyde fixative: a new fixative for immunoelectron microscopy. J Histochem Cytochem 22: 1077–1083
Streefkerk JG (1972) Inhibition of erythrocyte pseudoperoxidase activity by treatment with hydrogen peroxide following methanol. J Histochem Cytochem 20: 829–831
Barka, T, Anderson PJ (1962) Histochemical methods for acid phosphatase using hexazonium pararosanilin as coupler. J Histochem Cytochem 10: 741–753
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (Lond) 227: 680–685
Jee, WSS (1983) The skeletal tissues. In: Weiss L (ed) Histology, cell and tissue biology. the MacMillan Press, London, pp 200–255
Price PA, Otsuka AS, Poser JW, Kristaponis J, Raman N (1976) Characterization of a—carboxyglutamic acid-containing protein from bone. Proc Natl Acad Sci USA 73: 1447–1451
Romberg RW, Werness PG, Lollar P, Riggs BL, Mann KG (1985) Isolation and characterization of native adult osteonectin. J Biol Chem 260: 2728–2736
Lian JB, Tassinari M, Glowacki J (1984) Resorption of implanted bone prepared from normal and warfarin-treated rats. J Clin Invest 73: 1223–1226
Price PA, Williamson MK, Haba T, Dell RB, Jee WSS (1982) Excessive mineralization with growth plate closure in rats on chronic warfarin treatment. Proc Natl Acad Sci USA 79: 7734–7738
Mann K, Deutzmann R, Paulsson M, Timpl R (1987) Solubilization of protein BM-40 from a basement membrane tumor with chelating agents and evidence for its identity with osteonectin and SPARC. FEBS Lett 218: 167–172
Holland PWH, Harper SJ, McVey JH, Hogan BLM (1987) In vivo expression of mRNA for the Ca++-binding protein SPARC (osteonectin) revealed by in situ hybridization. J Cell Biol 105: 473–482
Malaval L, Fournier B, Delmas PD (1987) Radioimmunoassay for osteonectin. Concentrations in bone, nonmineralized tissues, and blood. J Bone Mineral Res 2: 457–465
Franzén A, Heinegård D (1985) Isolation and characterization of two sialoproteins present only in bone calcified matrix. Biochem J 232: 715–724
MacDougall M, Zeichner-David M, Slavkin HC (1985) Production and characterization of antibodies against murine dentin phosphoprotein. Biochem J 232: 493–500
Gorter de Vries I, Quartier E, Van Steirteghem A, Boute P, Coomans D, Wisse E (1986) Characterization and immunocytochemical localization of dentine phosphoprotein in rat and bovine teeth. Archs Oral Biol 31: 57–66
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Nakama, T., Nakamura, O., Daikuhara, Y. et al. A monoclonal antibody against dentin phosphophoryn recognizes a bone protein(s) appearing at the beginning of ossification. Calcif Tissue Int 43, 263–267 (1988). https://doi.org/10.1007/BF02555145
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02555145