Skip to main content
Log in

Expression of mRNA of murine bone-related proteins in ectopic bone induced by murine bone morphogenetic protein-4

  • Published:
Cell and Tissue Research Aims and scope Submit manuscript

Abstract

To determine whether a system of ectopic bone formation induced by osteosarcoma-derived bone-inducing substance (bone morphogenetic protein-4) can be used as a model of developing bone at the molecular level, we studied the expression of bone-related protein mRNAs in the process of ectopic bone formation using non-radioisotopic in situ hybridization. Osteonectin mRNA was detected in fibroblast-like cells, which are similar to periosteal cells from the early to middle stages of bone development. The proportion of osteonectin mRNA-expressing cells was greater than that of osteopontin mRNA-expressing cells in hypertrophic chondrocytes and osteoblast-like cells. In contrast, osteopontin mRNA was localized in a limited population of hypertrophic chondrocytes, a single layer of osteoblast-like cells adjacent to the bone trabeculae in the middle stage of bone formation, and in a limited subset of osteocytes in the late stage. A strong osteocalcin mRNA signal was detected in osteoblast-like cells from the middle to late stages and in a limited subset of osteocytes in the late stage of bone development. Since the sequential gene expression pattern of bone-related proteins in the present system is comparable to that in embryonic osteogenesis, this system may be useful as a model for studying gene expression in osteogenesis.

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

  • Amitani K, Nakata Y, Stevens J (1974) Bone induction by lyophilized osteosarcoma in mice. Calcif Tiss Res 16:305–313

    Google Scholar 

  • Bolander ME, Young MF, Fisher LW, Yamada Y, Termine JD (1988) Osteonectin cDNA sequencing reveals potential binding regions for calcium and hydroxyapatite, and shown homologies with both a basement membrane protein (SPARC) and a serine proteinase inhibitor(ovomucoid). Proc Natl Acad Sci USA 85:2919–2923

    Google Scholar 

  • Celeste AJ, Rosen V, Buecker JL, Kriz R, Wang EA, Wozney JM (1986) Isolation of the human gene for bone gla protein utilizing mouse and rat cDNA clones. EMBO J 5:1885–1890

    Google Scholar 

  • Craig AM, Smith JH, Denhardt DT (1989) Osteopontin, a transformation-associated cell adhesion phosphoprotein, is induced by 12-0-tetradecanoylphorbor 13-acetate in mouse epidermis. J Biol Chem 264:9682–9689

    Google Scholar 

  • Engel J, Taylor W, Paulsson M, Sage H, Hogan BLM (1987) Calcium binding domains and calcium-induced conformational transition of SPARC/BM-40/osteonectin, an extracellular glycoprotein expressed in mineralized and nonmineralized tissues. Biochemistry 26:6958–6965

    Google Scholar 

  • Franzen A, Heinegard D (1985) Isolation and characterization of two sialoproteins present only in bone calcified matrix. Biochem J 232:715–724

    Google Scholar 

  • Gundberg CM, Hauschka PV, Lian JB, Gallop PM (1984) Osteocalcin: isolation, characterization, and detection. Methods Enzymol 107:516–544

    Google Scholar 

  • Herring GM (1976) A comparison of bone matrix and tendon in particular reference to glycoprotein content. Biochem J 159:49–755

    Google Scholar 

  • Hirota S, Ito A, Morii E, Wanaka A, Tohyama M, Kitamura Y, Nomura S (1992) Localization of mRNA for c-kit receptor and its ligand in the brain of adult rats: an analysis using in situ hybridization histochemistry. Mol Brain Res 15:47–54

    Google Scholar 

  • Holland PWH, Harper S, 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

    Google Scholar 

  • Ikeda T, Nomura S, Yamaguchi A, Suda T, Yoshiki S (1992) In situ hybridization of bone matrix proteins in undecalcified adult rat bone sections. J Histochem Cytochem 40:1079–1088

    Google Scholar 

  • Mark MP, Prince CW, Oosawa T, Gay S, Bronckers ALJJ, Butler WT (1987a) Immunohistochemical demonstration of a 44kDal phosphoprotein in developing rat bone. J Histochem Cytochem 35:707–715

    Google Scholar 

  • Mark MP, Prince CW, Gay S, Austin RL, Bhown M, Finkelman RD, Butler WT (1987b) A comparative immunocytochemical study on the subcellular distributions of 44kDa bone phosphoprotein and bone gamma carboxyglutamic acid (Gla)-containing protein in osteoblasts. J Bone Miner Res 2:337–346

    Google Scholar 

  • Mark MP, Butler WT, Prince CW, Finkelman RD, Ruch JV (1988) Developmental expression of 44-kDa bone phosphoprotein (osteopontin) and bone gamma-carboxyglutamic acid (Gla)-containing protein (osteocalcin) in calcifying tissues of rat. Differentiation 37:123–136

    Google Scholar 

  • Mason JJ, Murphy D, Munke M, Francke U, Elliott RW, Hogan BLM (1986a)_Development and transformation-sensitive expression of the Sparc gene on mouse chromosome 11. EMBO J 5:1831–1837

    Google Scholar 

  • Mason IJ, Taylor A, Williams JG, Sage H, Hogan BLM (1986b) Evidence from molecular cloning that Sparc, a major product of mouse embryo parietal endoderm, is related to an endothelial cell ‘culture shock’ glycoprotein of Mr 43000. EMBO J 5:1465–1472

    Google Scholar 

  • Nomura S, Wills AJ, Edwards DR, Heath JK, Hogan BLM (1988) Developmental expression of 2ar (osteopontin) and SPARC (osteonectin) RNA as revealed by in situ hybridization. J Cell Biol 106:441–450

    Google Scholar 

  • Oldberg A, Franzen A, Heinegard 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

    Google Scholar 

  • Price PA, Otsuka AS, Poser JW, Kristaponis J, Raman N (1976) Characterization of a gamma-carboxyglutamic acid-containing protein from bone. Proc Natl Acad Sci USA 73:1447–1451

    Google Scholar 

  • Prince CW, Oosawa T, Butler WT, Tomana M, Bhown AS, Schrohenloher RE (1987) Isolation, characterization and biosynthesis of a phosphorylated glycoprotein from rat bone. J Biol Chem 262:2900–2907

    Google Scholar 

  • Sanger F, Nicklen S, Coulson A (1977) DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467

    Google Scholar 

  • Somerman MJ, Prince CW, Sauk JJ, Forsterman RA, Butler WT (1987) Mechanisms of fibroblast attachment to bone extracellular matrix: role of a 44 kilodalton bone phosphoprotein. J Bone Miner Res 2:259–265

    Google Scholar 

  • Takaoka K, Ono K, Amitami K, Kishimoto R, Nakano Y (1980) Solubilization and concentration of a bone-inducing substance from a murine osteosarcoma. Clin Orthop 148:274–280

    Google Scholar 

  • Takaoka K, Yoshikawa H, Hashimoto J, Miyamoto S, Masuhara K, Matsui M, Ono K (1993a) Purification and characterization of a bone-inducing protein from a murine osteosarcoma (Dunn type). Clin Orthop 292:329–336

    Google Scholar 

  • Takaoka K, Yoshikawa H, Hashimoto J, Masuhara K, Miyamoto S, Suzuki S, Ono K, Matsui M, Oikawa S, Tsuruoka N, Tawaragi Y, Inuzuka C, Katayama T, Sugiyama M, Tsujimoto M, Nakanishi T, Nakazato H (1993b) Gene cloning and expression of a bone morphogenetic protein derived from a murine osteosarcoma. Clin Orthop 294:344–352

    Google Scholar 

  • 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

    Google Scholar 

  • Theiler K (1989) The house mouse: atlas of embryonic development. Springer, New York

    Google Scholar 

  • Urist MR (1965) Bone formation by autoinduction. Science 150:893–899

    Google Scholar 

  • Young MF, Bolander ME, Day AA, Ramis CI, Robey PG, Yamada Y, Termine JD (1986) Osteonectin mRNA: distribution in normal and transformed cells. Nucleic Acids Res 14:4483–4497

    Google Scholar 

  • Weinreb M, Shinar D, Rodan GA (1990) Different pattern of alkaline phosphatase, osteopontin, and osteocalcin expression in developing rat bone visualized by in situ hybridization. J Bone Miner Res 5:831–842

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hirota, S., Takaoka, K., Hashimoto, J. et al. Expression of mRNA of murine bone-related proteins in ectopic bone induced by murine bone morphogenetic protein-4. Cell Tissue Res 277, 27–32 (1994). https://doi.org/10.1007/BF00303077

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00303077

Key words

Navigation