The SCPP gene repertoire in bony vertebrates and graded differences in mineralized tissues

  • Kazuhiko Kawasaki
Original Article


The vertebrate tooth is covered with enamel in most sarcopterygians or enameloid in chondrichthyans and actinopterygians. The evolutionary relationship among these two tissues, the hardest tissue in the body, and other mineralized tissues has long been controversial. We have recently reported that specific combinations of secretory calcium-binding phosphoprotein (SCPP) genes are involved in the mineralization of bone, dentin, enameloid, and enamel. Thus, the early repertoire of SCPP genes would elucidate the evolutionary relationship across these tissues. However, the diversity of SCPP genes in teleosts and tetrapods and the roles of these genes in distinct tissues have remained unclear, mainly because many SCPP genes are lineage-specific. In this study, I show that the repertoire of SCPP genes in the zebrafish, frog, and humans includes many lineage-specific genes and some widely conserved genes that originated in stem osteichthyans or earlier. Expression analysis demonstrates that some frog and zebrafish SCPP genes are used primarily in bone, but also in dentin, while the reverse is true of other genes, similar to some mammalian SCPP genes. Dentin and enameloid initially use shared genes in the matrix, but enameloid is subsequently hypermineralized. Notably, enameloid and enamel use an orthologous SCPP gene in the hypermineralization process. Thus, the hypermineralization machinery ancestral to both enameloid and enamel arose before the actinopterygian–sarcopterygian divergence. However, enamel employs specialized SCPPs as structuring proteins, not used in enameloid, reflecting the divergence of enamel from enameloid. These results show graded differences in mineralized dental tissues and reinforce the hypothesis that bone–dentin–enameloid–enamel constitutes an evolutionary continuum.


Vertebrate evolution Biomineralization Amelogenesis Odontogenesis Extracellular matrix protein 



I thank Prof. Keith C. Cheng and Ms. Peggy Hubley at Penn State University for providing me with zebrafish, Prof. Martin Flajnik and Dr. Yuko Ohta at the University of Maryland for the frog, and Dr. Chia-Lin Wei and Mr. Yow Jit Sin at the Genome Institute of Singapore for zebrafish clones. I am grateful to Prof. Kenneth M. Weiss and Dr. Anne V. Buchanan at Penn State University and Dr. Samuel Sholtis at Yale University for critical discussion and generous comments. This work was made possible by the financial support from awards SBR9804907 and BCS0343442 from the US National Science Foundation, and by research funds from Penn State University to Prof. Kenneth M. Weiss; and by NIH grant 5R24RR017441, and research grants from the Jake Gittlen Cancer Research Foundation and the Pennsylvania Tobacco Funds to Prof. Keith C. Cheng.

Supplementary material

427_2009_276_MOESM1_ESM.doc (32 kb)
ESM 1 (DOC 31.5 KB)


  1. Al Kawas S, Warshawsky H (2008) Ultrastructure and composition of basement membrane separating mature ameloblasts from enamel. Arch Oral Biol 53:310–317PubMedCrossRefGoogle Scholar
  2. Bartlett JD, Ganss B, Goldberg M, Moradian-Oldak J, Paine ML, Snead ML, Wen X, White SN, Zhou YL (2006) Protein–protein interactions of the developing enamel matrix. Curr Top Dev Biol 74:57–115PubMedCrossRefGoogle Scholar
  3. Bobe J, Goetz FW (2001) A novel osteopontin-like protein is expressed in the trout ovary during ovulation. FEBS Lett 489:119–124PubMedCrossRefGoogle Scholar
  4. Butler WT, Brunn JC, Qin C (2003) Dentin extracellular matrix (ECM) proteins: comparison to bone ECM and contribution to dynamics of dentinogenesis. Connect Tissue Res 44(Suppl 1):171–178PubMedCrossRefGoogle Scholar
  5. Currey JD (2002) Bones: structure and mechanics. Princeton University Press, PrincetonGoogle Scholar
  6. Donoghue PC, Sansom IJ, Downs JP (2006) Early evolution of vertebrate skeletal tissues and cellular interactions, and the canalization of skeletal development. J Exp Zoolog B Mol Dev Evol 306B:278–294CrossRefGoogle Scholar
  7. Fincham AG, Moradian-Oldak J, Simmer JP (1999) The structural biology of the developing dental enamel matrix. J Struct Biol 126:270–299PubMedCrossRefGoogle Scholar
  8. Fujisawa R, Butler WT, Brunn JC, Zhou HY, Kuboki Y (1993) Differences in composition of cell-attachment sialoproteins between dentin and bone. J Dent Res 72:1222–1226PubMedGoogle Scholar
  9. Hall BK (2005) Bones and cartilage: developmental and evolutionary skeletal biology. Elsevier, San DiegoGoogle Scholar
  10. Huysseune A, Van der heyden C, Sire JY (1998) Early development of the zebrafish (Danio rerio) pharyngeal dentition (Teleostei, Cyprinidae). Anat Embryol (Berl) 198:289–305CrossRefGoogle Scholar
  11. Iwasaki K, Bajenova E, Somogyi-Ganss E, Miller M, Nguyen V, Nourkeyhani H, Gao Y, Wendel M, Ganss B (2005) Amelotin—a novel secreted, ameloblast-specific protein. J Dent Res 84:1127–1132PubMedCrossRefGoogle Scholar
  12. Kawasaki K, Weiss KM (2003) Mineralized tissue and vertebrate evolution: the secretory calcium-binding phosphoprotein gene cluster. Proc Natl Acad Sci U S A 100:4060–4065PubMedCrossRefGoogle Scholar
  13. Kawasaki K, Weiss KM (2006) Evolutionary genetics of vertebrate tissue mineralization: the origin and evolution of the secretory calcium-binding phosphoprotein family. J Exp Zoolog B Mol Dev Evol 306B:295–316CrossRefGoogle Scholar
  14. Kawasaki K, Weiss KM (2007) Genetic basis for the evolution of vertebrate mineralized tissue. In: Bäuerlein E (ed) Handbook of biomineralization. Wiley-VCH, Weinheim, pp 331–347Google Scholar
  15. Kawasaki K, Weiss KM (2008) SCPP gene evolution and the dental mineralization continuum. J Dent Res 87:520–531PubMedCrossRefGoogle Scholar
  16. Kawasaki K, Shimoda S, Fukae M (1987) Histological and biochemical observations of developing enameloid of the sea bream. Adv Dent Res 1:191–195PubMedGoogle Scholar
  17. Kawasaki K, Suzuki T, Weiss KM (2004) Genetic basis for the evolution of vertebrate mineralized tissue. Proc Natl Acad Sci U S A 101:11356–11361PubMedCrossRefGoogle Scholar
  18. Kawasaki K, Suzuki T, Weiss KM (2005) Phenogenetic drift in evolution: the changing genetic basis of vertebrate teeth. Proc Natl Acad Sci U S A 102:18063–18068PubMedCrossRefGoogle Scholar
  19. Kawasaki K, Buchanan AV, Weiss KM (2007) Gene duplication and the evolution of vertebrate skeletal mineralization. Cells Tissues Organs 186:7–24PubMedCrossRefGoogle Scholar
  20. Laue K, Jänicke M, Plaster N, Sonntag C, Hammerschmidt M (2008) Restriction of retinoic acid activity by Cyp26b1 is required for proper timing and patterning of osteogenesis during zebrafish development. Development 135:3775–3787PubMedCrossRefGoogle Scholar
  21. Linde A, Goldberg M (1993) Dentinogenesis. Crit Rev Oral Biol Med 4:679–728PubMedGoogle Scholar
  22. Little EM, Holt C (2004) An equilibrium thermodynamic model of the sequestration of calcium phosphate by casein phosphopeptides. Eur Biophys J 33:435–447PubMedCrossRefGoogle Scholar
  23. Loh YH, Brenner S, Venkatesh B (2008) Investigation of loss and gain of introns in the compact genomes sof pufferfishes (fugu and tetraodon). Mol Biol Evol 25:526–535PubMedCrossRefGoogle Scholar
  24. Meinke DK (1982) A histological and histochemical study of developing teeth in Polypterus (Pisces, Actinopterygii). Arch Oral Biol 27:197–206PubMedCrossRefGoogle Scholar
  25. Meinke DK, Thomson KS (1983) The distribution and significance of enamel and enameloid in the dermal skeleton of osteolepiform. Paleobiology 9:138–149Google Scholar
  26. Moffatt P, Smith CE, Sooknanan R, St-Arnaud R, Nanci A (2006a) Identification of secreted and membrane proteins in the rat incisor enamel organ using a signal-trap screening approach. Eur J Oral Sci 114(Suppl 1):139–146PubMedCrossRefGoogle Scholar
  27. Moffatt P, Smith CE, St-Arnaud R, Simmons D, Wright JT, Nanci A (2006b) Cloning of rat amelotin and localization of the protein to the basal lamina of maturation stage ameloblasts and junctional epithelium. Biochem J 399:37–46PubMedCrossRefGoogle Scholar
  28. Moffatt P, Smith CE, St-Arnaud R, Nanci A (2008) Characterization of Apin, a secreted protein highly expressed in tooth-associated epithelia. J Cell Biochem 103:941–956PubMedCrossRefGoogle Scholar
  29. Nanci A, Zalzal S, Kogaya Y (1993) Cytochemical characterization of basement membranes in the enamel organ of the rat incisor. Histochemistry 99:321–331PubMedCrossRefGoogle Scholar
  30. Nanci A (2003) Ten Cate’s oral histology, 6th edn. Mosby, St. LouisGoogle Scholar
  31. Ng P, Wei CL, Sung WK, Chiu KP, Lipovich L, Ang CC, Gupta S, Shahab A, Ridwan A, Wong CH, Liu ET, Ruan Y (2005) Gene identification signature (GIS) analysis for transcriptome characterization and genome annotation. Nat Methods 2:105–111PubMedCrossRefGoogle Scholar
  32. Nieuwkoop PD, Faber J (1967) Normal table of Xenopus laevis (Daudin), 2nd edn. North-Holland, AmsterdamGoogle Scholar
  33. Paine ML, Snead ML (2005) Tooth developmental biology: disruptions to enamel–matrix assembly and its impact on biomineralization. Orthod Craniofac Res 8:239–251PubMedCrossRefGoogle Scholar
  34. Park JC, Park JT, Son HH, Kim HJ, Jeong MJ, Lee CS, Dey R, Cho MI (2007) The amyloid protein APin is highly expressed during enamel mineralization and maturation in rat incisors. Eur J Oral Sci 115:153–160PubMedCrossRefGoogle Scholar
  35. Poole DFG (1967) Phylogeny of tooth tissues: Enameloid and enamel in recent vertebrates with a note in the history of cementum. In: Miles AEW (ed) Structural and chemical organization of teeth. Academic, New York, pp 111–149Google Scholar
  36. Poole DFG (1971) An introduction to the phylogeny of calcified tissues. In: Dahlberg AA (ed) Dental morphology and evolution. University of Chicago Press, Chicago, pp 65–79Google Scholar
  37. Qin C, Brunn JC, Jones J, George A, Ramachandran A, Gorski JP, Butler WT (2001) A comparative study of sialic acid-rich proteins in rat bone and dentin. Eur J Oral Sci 109:133–141PubMedCrossRefGoogle Scholar
  38. Qin C, Brunn JC, Cadena E, Ridall A, Tsujigiwa H, Nagatsuka H, Nagai N, Butler WT (2002) The expression of dentin sialophosphoprotein gene in bone. J Dent Res 81:392–394PubMedCrossRefGoogle Scholar
  39. Qin C, Baba O, Butler WT (2004) Post-translational modifications of sibling proteins and their roles in osteogenesis and dentinogenesis. Crit Rev Oral Biol Med 15:126–136PubMedCrossRefGoogle Scholar
  40. Qin C, D’Souza R, Feng JQ (2007) Dentin matrix protein 1 (DMP1): new and important roles for biomineralization and phosphate homeostasis. J Dent Res 86:1134–1141PubMedCrossRefGoogle Scholar
  41. Rijnkels M, Elnitski L, Miller W, Rosen JM (2003) Multispecies comparative analysis of a mammalian-specific genomic domain encoding secretory proteins. Genomics 82:417–432PubMedCrossRefGoogle Scholar
  42. Sasagawa I (1997) Fine structure of the cap enameloid and of the dental epithelial cells during enameloid mineralisation and early maturation stages in the tilapia, a teleost. J Anat 190(Pt 4):589–600PubMedCrossRefGoogle Scholar
  43. Sasagawa I, Ishiyama M (1988) The structure and development of the collar enameloid in two teleost fishes, Halichoeres poecilopterus and Pagrus major. Anat Embryol (Berl) 178:499–511CrossRefGoogle Scholar
  44. Shellis RP, Miles AEW (1974) Autoradiographic study of the formation of enameloid and dentine matrices in teleost fishes using tritiated amino acids. Proc R Soc Lond B 185:51–72CrossRefGoogle Scholar
  45. Shintani S, Kobata M, Toyosawa S, Ooshima T (2003) Identification and characterization of ameloblastin gene in an amphibian, Xenopus laevis. Gene 318:125–136PubMedCrossRefGoogle Scholar
  46. Simmer JP, Fincham AG (1995) Molecular mechanisms of dental enamel formation. Crit Rev Oral Biol Med 6:84–108PubMedCrossRefGoogle Scholar
  47. Simmer JP, Hu JC (2002) Expression, structure, and function of enamel proteinases. Connect Tissue Res 43:441–449PubMedCrossRefGoogle Scholar
  48. Smith CE (1998) Cellular and chemical events during enamel maturation. Crit Rev Oral Biol Med 9:128–161PubMedCrossRefGoogle Scholar
  49. Smith MM (1992) Microstructure and evolution of enamel amongst osteichthyan and early tetrapods. In: Smith P, Tchernov E (eds) Structure, function and evolution of teeth. Freund, Tel Aviv, pp 73–101Google Scholar
  50. Smith MM (1995) Heterochrony in the evolution of enamel in vertebrates. In: McNamara KJ (ed) Evolutionary change and heterochrony. Wiley, Chichester, pp 125–150Google Scholar
  51. Smyth E, Clegg RA, Holt C (2004) A biological perspective on the structure and function of caseins and casein micelles. Int J Dairy Technol 57:121–126CrossRefGoogle Scholar
  52. Sodek J, Ganss B, McKee MD (2000) Osteopontin. Crit Rev Oral Biol Med 11:279–303PubMedCrossRefGoogle Scholar
  53. Solomon A, Murphy CL, Weaver K, Weiss DT, Hrncic R, Eulitz M, Donnell RL, Sletten K, Westermark G, Westermark P (2003) Calcifying epithelial odontogenic (Pindborg) tumor-associated amyloid consists of a novel human protein. J Lab Clin Med 142:348–355PubMedCrossRefGoogle Scholar
  54. Stock DW (2007) Zebrafish dentition in comparative context. J Exp Zoolog B Mol Dev Evol 308:523–549PubMedCrossRefGoogle Scholar
  55. Takano Y (1979) Cytochemical studies of ameloblasts and the surface layer of enamel of the rat incisor at the maturation stage. Arch Histol Jpn 42:11–32PubMedGoogle Scholar
  56. Toyosawa S, O’hUigin C, Figueroa F, Tichy H, Klein J (1998) Identification and characterization of amelogenin genes in monotremes, reptiles, and amphibians. Proc Natl Acad Sci U S A 95:13056–13061PubMedCrossRefGoogle Scholar
  57. Toyosawa S, Shintani S, Fujiwara T, Ooshima T, Sato A, Ijuhin N, Komori T (2001) Dentin matrix protein 1 is predominantly expressed in chicken and rat osteocytes but not in osteoblasts. J Bone Miner Res 16:2017–2026PubMedCrossRefGoogle Scholar
  58. Van der heyden C, Huysseune A (2000) Dynamics of tooth formation and replacement in the zebrafish (Danio rerio) (Teleostei, Cyprinidae). Dev Dyn 219:486–496PubMedCrossRefGoogle Scholar
  59. Van der heyden C, Huysseune A, Sire JY (2000) Development and fine structure of pharyngeal replacement teeth in juvenile zebrafish (Danio rerio) (Teleostei, Cyprinidae). Cell Tissue Res 302:205–219CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  1. 1.Department of AnthropologyPennsylvania State UniversityUniversity ParkUSA

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