Advertisement

Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Rat enamel contains RP59: a new context for a protein from osteogenic and haematopoietic precursor cells

  • 61 Accesses

  • 2 Citations

Abstract

We have recently identified a protein, RP59, in bone marrow cells and young osteoblasts, in cells involved in bone repair and in young erythroblasts and megakaryocytes. Here, we report immunohistochemical data at the light- and electron-microscope level indicating that RP59 is also present in newly secreted tooth enamel of the rat and in ameloblasts, the formative cells. In enamel matrix, RP59 was located proximal to secretory ameloblasts only, i.e. in newly secreted material. Distal enamel and enamel in association with maturation stage ameloblasts were unlabelled. Secretory ameloblasts contained RP59 in the matrix-proximal region including Tomes’ processes, post-secretory ameloblasts in the cell-matrix interface. Western blotting of proteins from tooth germs identified RP59 as a band at 90 kD, co-migrating with RP59 from bone marrow and spleen. Antisera versus a chemically synthesised RP59 peptide and versus a bacteria-synthesised protein fragment reacted in the same manner. In situ hybridisation of tooth tissue revealed RP59 RNA specifically in ameloblasts. The reverse transcription/polymerase chain reaction method identified tooth RNA coding for RP59. Sequence analysis indicated that RP59 RNA from tooth and marrow had the same sequence. An internal sequence motif was found in rat RP59 resembling a signal implicated in secretion of the chicken “engrailed” gene product. The findings indicate that RP59 is a genuine product of ameloblasts and that it is secreted in the course of enamel formation together with other matrix components.

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

References

  1. Bartlett JD, Simmer JP (1999) Proteinases in developing dental enamel. Crit Rev Oral Biol Med 10:425–441

  2. Begue-Kirn C, Krebsbach PH, Bartlett JD, Butler WT (1998) Dentin sialoprotein, dentin phosphoprotein, enamelysin and ameloblastin: tooth-specific molecules that are distinctively expressed during murine dental differentiation. Eur J Oral Sci 106:963–970

  3. Cerny R, Slaby I, Hammarstrom L, Wurtz T (1996) A novel gene expressed in rat ameloblasts codes for proteins with cell binding domains. J Bone Miner Res 11:883–891

  4. Chen Y-C, Wurtz T, Wang C-J, Kuo Y-R, Yang KD, Huang H-C, Wang F-S (2004) Recruitment of mesenchymal stem cells and expression of TGF-β 1 and VEGF in the early stage of shock wave-promoted bone regeneration of segmental defect in rats. J Orthoped Res 22:526–534

  5. Deutsch D, Haze-Filderman A, Taylor AL, Blumenfeld A, Dadni L, Shay B, Mao Z, Gruenbaum-Cohen Y, Rosenfeld E, Leiser Y, Fermon E, Shai E, Jawor P, Zimmermann B, Haegewald S, Bernimoulin JP (2004) Amelogenin in cementum, periodontal ligament and bone cells in normal and regenerating tissues (contribution to 8th International Conference on Tooth Morphogenesis and Differentiation)

  6. Downey LM, Keen TJ, Jalili IK, McHale J, Aldred MJ, Robertson S, Mighell A, Fayle S, Wissinger B, Inglehearn CF (2002) Identification of a locus on chromosome 2q11 at which recessive amelogenesis imperfecta and cone-rod dystrophy cosegregate. Eur J Hum Genet 10:865–869

  7. Fong CD, Cerny R, Hammarström L, Slaby I (1998) Sequential expression of an amelin gene in mesenchymal and epithelial cells during odontogenesis in rats. Eur J Oral Sci 106 (Suppl 1):324–330

  8. Fraser AG, Marcotte EM (2004) A probabilistic view of gene function. Nat Genet 36:559–564

  9. Gama-Carvalho M, Carmo-Fonseca M (2001) The rules and roles of nucleocytoplasmic shuttling proteins. FEBS Lett 498:157–163

  10. Gibson C, Yuan Z-A, Hall B, Longenecker G, Chen E, Thyagarajan T, Sreenath T, Wright JT, Decker S, Piddington R, Harrison G, Kulkarni AB (2001) Amelogenin deficient mice display an amelogenesis imperfecta phenotype. J Biol Chem 34:31871–31875

  11. Goldberg M, Septier D, Bourd K, Hall R, Jeanny JC, Jonet L, Colin S, Tage F, Chaussain-Miller C, Garabedian M., George A, Goldberg H, Menashi S (2002) The dentino-enamel junction revisited. Connect Tissue Res 43:482–489

  12. Goldberg M, Six N, Decup F, Lasfargues J-J, Salih E, Tompkins K, Veis A (2003) Bioactive molecules and the future of pulp therapy. Am J Dent 16:66–76

  13. Haga A, Niinaka Y, Raz A (2000) Phosphohexose isomerase/autocrine motility factor/ neuroleukin/maturation factor is a multifunctional phosphoprotein. Biochem Biophys Acta 1480:235–244

  14. Hart TC, Marazita ML, Wright JT (2000) The impact of molecular genetics on oral health paradigms. Crit Rev Oral Biol Med 11:6–56

  15. Hu CC, Fukae M, Uchida T, Quian Q, Zhang CH, Ryu OH, Tanabe T, Yamakoshi Y, Murakami C, Dohi N, Shimizu M, Simmer JP (1997) Cloning and characterization of porcine enamelin mRNAs. J Dent Res 76:1720–1729

  16. Hultenby K, Reinholt FP, Oldberg Å, Heinegård D (1991) Ultrastructural immunolocalization of osteopontin in metaphyseal and cortical bone. Matrix 11:206–213

  17. Hultenby K, Reinholt FP, Norgård M, Oldberg Å, Wendel M, Heinegård D (1994) Distribution and synthesis of bone sialoprotein in metaphyseal bone of young rats show a distinctly different pattern from that of osteopontin. Eur J Cell Biol 62:230–239

  18. Jeffery CJ (2003) Moonlighting proteins: old proteins learning new tricks. Trends Genet 19:415–417

  19. Joliot A, Maizel A, Rosenberg D, Trembleau A, Dupas S, Volovitch M, Prochiantz A (1998) Identification of a signal sequence necessary for the unconventional secretion of Engrailed homeoprotein. Curr Biol 8:856–863

  20. Krebsbach PH, Lee SK, Matsuki Y, Kozak CA, Yamada KM, Yamada Y (1996) Full-length sequence, localization, and chromosomal mapping of ameloblastin. A novel tooth-specific gene. J Biol Chem 271:4431–4435

  21. Krüger A, Ellerström C, Lundmark C, Christersson C, Wurtz T (2002) RP59, a marker for osteoblast recruitment, is also detected in primitive mesenchymal cells, erythroid cells and megakaryocytes. Dev Dyn 233:414–418

  22. Lagerström M, Dahl N, Nakahori Y, Nakagome Y, Bäckman B, Landegren U, Petterson U (1991) A deletion in the amelogenin gene (AMG) causes X-linked amelogenesis imperfecta (AIH1). Genomics 10:971–975

  23. Lezot F, Davideau JL, Thomas B, Sharpe P, Forest N, Berdal A (2000) Epithelial Dlx-2 homeogene expression and cementogenesis. J Histochem Cytochem 48:277–284

  24. Li H, Oberhauser AF, Redick SD, Carrion-Vasquez M, Erickson HP, Fernandez JM (2001) Multiple conformations of PEVK proteins detected by single molecule techniques. Proc Nat Acad Sci USA 98:10682–10686

  25. Mårdh CK, Bäckman B, Holmgren G, Hu JC, Simmer JP, Forsman-Semb K (2002) A nonsense mutation in the enamelin gene causes local hypoplastic autosomal dominant amelogenesis imperfecta (AIH2). Hum Mol Genet 11:1069–1074

  26. Nanci A, Zalzal S, Lavoi P, Kunikata M, Che W-Y, Krebsbach PH, Yamada Y, Hammarström L, Simmer JP, Fincham AG, Snead ML, Smith CE (1998) Comparative immunochemical analyses of the developmental expression and distribution of ameloblastin and amelogenin in rat incisors. J Histochem Cytochem 46:911–934

  27. Nickel W (2003) The mystery of nonclassical protein secretion. Eur J Biochem 270:2109–2119

  28. Orkin SH, Zon LI (2002) Hematopoiesis and stem cells: plasticity versus developmental heterogeneity. Nat Immunol 3:323–328

  29. Paine ML, Wang HJ, Luo W, Krebsbach PH, Snead ML (2003) A transgenic animal model resembling amelogenesis imperfecta related to ameloblastin over-expression. J Biol Chem 278:19447–19452

  30. Priam F, Ronco V, Bourd, Lamblin D, Bonnefoix M, Duchêne T, Locker M, Wurtz T, Kellermann O, Goldberg M, Poliard A (2005) New cellular models for tracking the odontoblast phenotype. Arch Oral Biol (in press)

  31. Rajpar MH, Harley K, Laing C, Davies RM, Dixon MJ (2001) Mutation of the gene encoding the enamel-specific protein enamelin causes autosomal-dominant amelogenesis imperfecta. Hum Mol Genet 10:1673–1677

  32. Reinholt F, Hultenby K, Heinegard D, Marks SC Jr, Norgard M, Anderson G (1999) Extensive clear zone and defective ruffled border formation in osteoclasts of osteopetrotic (ia/ia) rats: implications for secretory function. Exp Cell Res 251:477–491

  33. Robinson C, Brookes SJ, Shore RC, Kirkham J (1998) The developing enamel matrix: nature and function. Eur J Oral Sci 106 (Suppl 1):282–291

  34. Simmer JP, Fincham AG (1995) Molecular mechanisms of dental enamel formation. Crit Rev Oral Biol Med 6:84–108

  35. Uchida T, Murakami C, Dohi N, Wakida K, Satoda T, Takahashi O (1997) Synthesis, secretion, degradation and fate of ameloblastin during the matrix formation state of the rat incisor as shown by immunocytochemistry and immunochemistry using region-specific antibodies. J Histochem Cytochem 45:1329–1340

  36. Wilkinson DG, Green J (1990) In situ hybridization and the three-dimensional reconstruction of serial sections: postimplantation mammalian embryos. Oxford University press, London, pp 155–171

  37. Wurtz T, Berdal A (2003) Osteoblast precursors at different anatomic sites. Crit Rev Eukaryot Gene Expr 13:147–161

  38. Wurtz T, Krüger A, Christersson C, Lundmark C (2001) A new protein expressed in bone marrow cells and osteoblasts with implication in osteoblast recruitment. Exp Cell Res 263:236–242

  39. Yang LV, Nicholson RH, Kaplan J, Galy A, Li L (2001) Hemogen is a novel nuclear factor specifically expressed in mouse hematopoietic development and its human homologue EDAG maps to chromosome 9q22, a region containing breakpoints of hematological neoplasms. Mech Dev 104:105–111

Download references

Acknowledgements

We thank Ingrid Lindell for technical assistance.

Author information

Correspondence to T. Wurtz.

Additional information

A. Krüger and E. Somogyi contributed equally to this work.

This work was funded by the Stockolm County Council and by a grant from the Medical Research Council (Sweden) to T.W.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Krüger, A., Somogyi, E., Christersson, C. et al. Rat enamel contains RP59: a new context for a protein from osteogenic and haematopoietic precursor cells. Cell Tissue Res 320, 141–148 (2005). https://doi.org/10.1007/s00441-004-1043-y

Download citation

Keywords

  • Enamel formation
  • Tooth development
  • Ameloblasts
  • Extracellular matrix
  • PEVK-rich proteins
  • Rat (Sprague Dawley)