Skip to main content
SpringerLink
Log in

Immunohistochemical detection of an enamel protein-related epitope in rat bone at an early stage of osteogenesis

  • Originals
  • Published:
Histochemistry Aims and scope Submit manuscript

Abstract

Monoclonal antibody MI315 was produced against hamster tooth germ homogenate by in vitro immunization. It was found that MI315 reacted with enamel matrix, ameloblasts, and bone matrix at an early stage of osteogenesis. Decalcified tissues of rat femurs and mandibles were examined with MI315 using indirect immunofluorescence. In endochondral ossification of femurs, immunoreactivity was found in bone extracellular matrix (ECM) deposited on the surface of the cartilage core of primary spongiosa, but not in the cartilage core itself. In intramembranous ossification of 0-day-old rat mandibles, intense immunofluorescence was detected in bone ECM and a few young osteocytes, but not in osteoblasts. Immunoreactivity in bone ECM of 2-day-old rats decreased and almost disappeared from bone ECM of 4-day-old rats. Although in nondecalcified sections of 0-day-old rats, negligible immunofluorescence was detected in bone ECM which showed positive staining in decalcified tissues, the immunostaining appeared after decalcification using ethylenediaminetetraacetic acid (EDTA). These results indicate that a substance(s), which had a common epitope with an enamel-derived protein(s), existed in immature bone ECM of both endochondral and intramembranous ossification, and that it might be masked by bone mineral. Monoclonal antibody MI315 is a useful tool to investigate the time- and position-specific changes in osteogenesis and amelogenesis.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Akita H, Kobayashi Y, Kagayama M (1988) A histochemical study on lectin binding in the immature enamel and secretory ameloblasts of rat incisors. Tohoku J Exp Med 155:139–149

    Google Scholar 

  • Bianco P (1990) Ultrastructural immunohistochemistry of noncollagenous proteins in calcified tissues. In: Bonucci E, Motta PM (eds) Ultrastructure of skeletal tissues. Bone and cartilage in health and disease. Kluwer, Boston, pp 63–78

    Google Scholar 

  • Boss BD (1984) An improved in vitro immunization procedure for the production of monoclonal antibodies against neural and other antigens. Brain Res 291:193–196

    Google Scholar 

  • Bronckers ALJ, Gay S, Dimuzio MT, Butler WT (1985) Immunolocalization of γ-carboxyglutamic acid containing proteins in developing rat bones. Collagen Rel Res 5:273–281

    Google Scholar 

  • Bronckers ALJJ, Lyaruu DM, Wöltgens JHM (1989) Immunohistochemistry of extracellular matrix proteins during various stages of dentinogenesis. Connect Tissue Res 22:65–70

    Google Scholar 

  • Burgess RC, MacLaren (1965) Proteins in developing bovine enamel. In: Fearnhead RW, Stack MV (eds) Tooth enamel. Wright, Bristol, pp 74–82

    Google Scholar 

  • Burnell JM, Teubner EJ, Miller AG (1980) Normal maturational changes in bone matrix, mineral, and crystal size in the rat. Calcif Tissue Int 31:13–19

    Google Scholar 

  • Butler WT (1984) Matrix macromolecules of bone and dentin. Collagen Rel Res 4:297–307

    Google Scholar 

  • Butler WT (1991) Sialoproteins of bone and dentin. J Biol Buccale 19:83–89

    Google Scholar 

  • Camarda AJ, Butler WT, Finkelman RD, Nanci A (1987) Immunocytochemical localization of γ-carboxyglutamic acid-containing proteins (osteocalcin) in rat bone and dentin. Calcif Tissue Int 40:349–355

    Google Scholar 

  • Deakins M (1942) Changes in the ash, water and organic content of pig enamel during calcification. J Dent Res 21:429–435

    Google Scholar 

  • Eastoe JE (1963) The amino acid composition of proteins from the oral tissues. II. The matrix proteins in dentine and enamel from developing human deciduous teeth. Arch Oral Biol 8:633–652

    Google Scholar 

  • Eastoe JE (1979) Enamel protein chemistry — Past, present and future. J Dent Res 58B:735–763

    Google Scholar 

  • Eastoe JE, Camilleri GE (1971) The pattern of protein distribution in enamel during maturation. In: Fearnhead RW, Stack MV (eds) Tooth enamel II. Wright, Bristol, pp 119–141

    Google Scholar 

  • Fearnhead RW (1979) Matrix-mineral relationships in enamel tissues. J Dent Res 58B:909–916

    Google Scholar 

  • Fincham AG (1982) The extracellular matrix of developing dental enamel. In: Anderson W, Sadler W (eds) Perspective in differentiation and hypertrophy. Elsevier, New York, pp 135–156

    Google Scholar 

  • Graver HT, Herold RC, Chung T-Y, Christner PJ, Pappas C, Rosenbloom J (1978) Immunofluorescent localization of amelogenins in developing bovine teeth. Dev Biol 63:390–401

    Google Scholar 

  • Grynpas MD, Hunter GK (1988) Bone mineral and glycosaminoglycans in newborn and mature rabbits. J Bone Mineral Res 3:159–164

    Google Scholar 

  • Herold RC, Boyde A, Rosenbloom J, Lally ET (1987) Monoclonal antibody and immunogold cytochemical localization of amelogenins in bovine secretory amelogenesis. Arch Oral Biol 32:439–444

    Google Scholar 

  • Inage T, Shimokawa H, Teranishi Y, Iwase T, Toda Y, Moro I (1989) Immunocytochemical demonstration of amelogenins and enamelins secreted by ameloblasts during the secretory and maturation stages. Arch Histol Cytol 52:213–229

    Google Scholar 

  • Inai T, Kukita T, Ohsaki Y, Nagata K, Kukita A, Kurisu K (1991) Immunohistochemical demonstration of amelogenin penetration toward the dental pulp in the early stages of ameloblast development in rat molar tooth germs. Anat Rec 229:259–270

    Google Scholar 

  • Inai T, Nagata K, Kukita T, Kurisu K (1992) Demonstration of amelogenin in the enamel-free cusps of rat molar tooth germs: Immunofluorescent and immunoelectron microscopic studies. Anat Rec 233:588–596

    Google Scholar 

  • Johnson GD, Araujo GMCN (1981) A simple method of reducing the fading of immunofluorescence during microscopy. J Immunol Methods 43:349–350

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of the bacteriophage T4. Nature 227:680–685

    Google Scholar 

  • Lesot H, Smith AJ, Matthews JB, Ruch JV (1988) An extracellular matrix protein of dentine, enamel, and bone shares common antigenic determinants with keratins. Calcif Tissue Int 42:53–57

    Google Scholar 

  • Limeback H, Sakarya H, Chu W, Mackinnon M (1989) Serum albumin and its acid hydrolysis peptides dominate preparations of mineral-bound enamel proteins. J Bone Mineral Res 4:235–241

    Google Scholar 

  • Linde A (1989) Dentine matrix proteins: composition and possible functions in calcification. Anat Rec 224:154–166

    Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275

    Google Scholar 

  • Luk SC, Nopajaroonsri C, Simon GT (1974) The ultrastructure of cortical bone in young adult rabbits. J Ultrastruct Res 46:184–205

    Google Scholar 

  • Mackie EJ, Thesleff I, Chiquet-Ehrismann R (1987) Tenascin is associated with chondrogenic and osteogenic differentiation in vivo and promotes chondrogenesis in vitro. J Cell Biol 105:2569–2579

    Google Scholar 

  • McLean IW, Nakane PK (1974) Periodate-lysine-paraformaldehyde fixative, a new fixative for immunoelectron microscopy. J Histochem Cytochem 22:1077–1083

    Google Scholar 

  • Nakamura M, Akita H, Mizoguchi I, Kagayama M (1989) A histochemical localization onMaclura pomifera lectin during osteogenesis. Histochemistry 92:225–230

    Google Scholar 

  • Nanci A, Warshawsky H (1984) Relationship between the quality of fixation and the presence of stippled material in newly formed enamel of the rat incisor. Anat Rec 208:15–31

    Google Scholar 

  • Nanci A, Bendayan M, Slavkin HC (1984) Distribution of enamel protein antigens during mouse incisor amelogenesis as revealed by high resolution immunocytochemistry. In: Fearnhead RW, Suga S (eds) Tooth enamel IV. Elsevier, Amsterdam, pp 141–145

    Google Scholar 

  • Nanci A, Bendayan M, Slavkin HC (1985) Enamel protein biosynthesis and secretion in mouse incisor secretory ameloblasts as revealed by high-resolution immunocytochemistry. J Histochem Cytochem 33:1153–1160

    Google Scholar 

  • Nanci A, Slavkin HC, Smith CE (1987) Immunocytochemical and radioautographic evidence for secretion and intracellular degradation of enamel proteins by ameloblasts during the maturation stage of amelogenesis in rat incisors. Anat Rec 217:107–123

    Google Scholar 

  • Nanci A, Ahluwalia JP, Pompura JR, Smith CE (1989) Biosynthesis and secretion of enamel proteins in the rat incisors. Anat Rec 224:227–291

    Google Scholar 

  • Ohnishi T, Arakaki N, Nakamura O, Hirono S, Daikuhara Y (1991) Purification, characterization, and studies on biosynthesis of a 59-kDa bone sialic acid-containing protein (BSP) from rat mandible using a monoclonal antibody. J Biol Chem 266:14636–14645

    Google Scholar 

  • Pardue RL, Brady RC, Perry GW, Dedman JR (1983) Production of monoclonal antibodies against calmodulin by in vitro immunization of spleen cells. J Cell Biol 96:1149–1154

    Google Scholar 

  • Reading CL (1982) Theory and methods for immunization in culture and monoclonal antibody production. J Immunol Methods 53:261–291

    Google Scholar 

  • Sasaki T (1990a) Golgi-GERL-lysosome systems and resorptive function. In: Sasaki T (ed) Cell biology of tooth enamel formation. Karger, Basel, pp 62–65

    Google Scholar 

  • Sasaki T (1990b) Resorptive function of maturation ameloblasts. In: Sasaki T (ed) Cell biology of tooth enamel formation. Karger, Basel, pp 121–124

    Google Scholar 

  • Slavkin HC, Zeichner-David M, Siddique MAQ (1981) Molecular aspects of tooth morphogenesis and differentiation. Mol Aspects Med 4:125–188

    Google Scholar 

  • Slavkin HC, Zeichner-David M, MacDougal M, Bringas P, Bessem C, Honing LS (1982) Antibodies to murine amelogenins: localization of enamel proteins during tooth organ development in vitro. Differentiation 23:73–82

    Google Scholar 

  • Slavkin HC, Bessem C, Bringas P, Zeichner-David M, Nanci A, Snead ML (1988a) Sequential expression and differential function of multiple enamel proteins during fetal, neonatal, and early postnatal stages of mouse molar organogenesis. Differentiation 37:26–39

    Google Scholar 

  • Slavkin HC, Bringas P Jr, Bessem C, Santos V, Nakamura M, Hsu M-y, Snead ML, Zeichner-David M, Fincham AG (1988b) Hertwich's epithelial root sheath differentiation and initial cementum and bone formation during long-term organ culture of mouse mandibular first molars using serumless, chemically-defined medium. J Periodont Res 23:28–40

    Google Scholar 

  • Slavkin HC, Bessem C, Fincham AG, Bringas P, Santos V Jr, Snead ML, Zeichner-David M (1989) Human and mouse cementum proteins immunologically related to enamel proteins. Biochim Biophys Acta 991:12–18

    Google Scholar 

  • Stack MV (1967) Chemical organization of the organic matrix of enamel. In: Miles AEW (ed) Structural and chemical organization of teeth, vol 2. Academic Press, New York, pp 317–345

    Google Scholar 

  • Takagi Y, Fujisawa R, Sasaki S (1986) Identification of dentin phosphophoryn localization by histochemical stainings. Connect Tissue Res 14:279–292

    Google Scholar 

  • Termine JD (1988) Non-collagen proteins in bone. In: Evered D, Harnett S (eds) Cell and molecular biology of vertebrate hard tissues, Ciba Foundation Symposium 136. Wiley, Chichester, pp 178–190

    Google Scholar 

  • Termine JD, Belcourt AB, Christner PJ, Conn KM, Nylen MU (1980) Properties of dissociatively extracted fetal tooth matrix proteins. I. Principal molecular species in developing bovine enamel. J Biol Chem 255:9760–9768

    Google Scholar 

  • Thesleff I, Kantomaa T, Mackie E, Chiquet-Ehrismann R (1988) Immunohistochemical localization of the matrix glycoprotein tenascin in the skull of the growing rat. Arch Oral Biol 33:383–390

    Google Scholar 

  • Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Second Department of Conservative Dentistry, Faculty of Dentistry, Kyushu University 61, Maidashi, Higashi-Ku, 812, Fukuoka, Japan

    Tetsuichiro Inai

  2. Department of Preventive Dentistry, Faculty of Dentistry, Kyushu University 61, Maidashi, Higashi-Ku, 812, Fukuoka, Japan

    Yuko Inai

  3. First Department of Anatomy, Osaka University Faculty of Dentistry, Osaka, Japan

    Kojiro Kurisu

Authors
  1. Tetsuichiro Inai
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yuko Inai
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Kojiro Kurisu
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Inai, T., Inai, Y. & Kurisu, K. Immunohistochemical detection of an enamel protein-related epitope in rat bone at an early stage of osteogenesis. Histochemistry 99, 355–362 (1993). https://doi.org/10.1007/BF00717047

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00717047

Keywords

Search

Navigation