International Journal of Oral Biology

  • 제43권4호
  • /
  • Pages.177-183
  • /
  • 2018
  • /
  • 1226-7155(pISSN)
  • /
  • 2287-6618(eISSN)
대한구강생물학회 (The Korean Academy of Oral Biology)
권호 표지
DOI QR코드

DOI QR Code

Expression and Localization of Keap1 During Amelogenesis in the Developing Molar Germ of Rats

  • Kim, Sun-Hun (Dental Science Research Institute, Chonnam National University) ;
  • You, Yong-Ouk (Institute of Biomaterial.Implant, Department of Oral Biochemistry, School of Dentistry, Wonkwang University) ;
  • Ko, Hyun-Mi (Dental Science Research Institute, Chonnam National University) ;
  • Kim, Hyun-Jin (Institute of Biomaterial.Implant, Department of Oral Anatomy, School of Dentistry, Wonkwang University)
  • 투고 : 2018.10.15
  • 심사 : 2018.12.10
  • 발행 : 2018.12.31
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

The objective of this study was to examine the expression pattern of Kelch-like ECH-associated protein 1 (Keap1) in the maxillary $2^{nd}$ molar germs of rats. We used the maxillary $2^{nd}$ molar germs in rats' pup at postnatal day 3 (bell stage), 6 (crown formation stage) and 9 (root formation stage). The investigation on mRNA and protein levels were done using reverse transcription - polymerase chain reaction and western blot. Localization of Keap 1 in the maxillary $2^{nd}$ molar germs were revealed through immunofluorescence staining. Keap1 from the maxillary 2nd molar germs were mostly manifested on postnatal day 3 and dramatically decreased on postnatal day 6 and 9 at mRNA and protein levels, while amelogenin and ameloblastin increased during the development of maxillary 2nd molar germs. During immunofluorescence analysis, the strong immunoreactivity against Keap1 was detected in the apical side of ameloblasts at the presecretory and secretory stages. However, Keap1 expression was hardly observed in the ameloblasts at the maturation stage. These results shows that Keap1 is strongly expressed in the presecretory and secretory ameloblasts of amelogenesis, and suggest that Keap1 may be a crucial molecule for the regulatory mechanisms tasked with the formation of enamel layer.

키워드

참고문헌

  1. Kollar EJ, Mina M. Role of the early epithelium in the patterning of the teeth and Meckel’s cartilage. J Craniofac Genet Dev Biol. 1991;11:223-228.
  2. Slavkin HC. Molecular determinants during dental morphogenesis and cytodifferentiation: a review. J Craniofac Gen Dev Biol. 1991;11:338-349.
  3. Thesleff I. Epithelial–mesenchymal signaling regulating tooth morphogenesis. J Cell Sci. 2003;116:1647-1648. https://doi.org/10.1242/jcs.00410
  4. Pispa J, Thesleff I. Mechanisms of ectodermal organogenesis. Dev Biol. 2003;262:195-205. https://doi.org/10.1016/S0012-1606(03)00325-7
  5. Simmer JP1, Papagerakis P, Smith CE, Fisher DC, Rountrey AN, Zheng L, Hu JC. Regulation of dental enamel shape and hardness. J Dent Res. 2010;89:1024-1038. doi: 10.1177/0022034510375829.
  6. Fincham AG, Moradian-oldak J, Simmer JP. The structural biology of the developing dental enamel metrix. J Struct Biol. 1999;126:270-299. https://doi.org/10.1006/jsbi.1999.4130
  7. Moradian-Oldak J. Amelogenins: assembly, processing and control of crystal morphology. Matrix Biol. 2001;20: 293-305. https://doi.org/10.1016/S0945-053X(01)00154-8
  8. Cerny R, Slaby I, Hammarström L, Wurtz T. A novel gene expressed in rat ameloblasts codes for proteins with cell binding domains. J Bone Miner Res. 1996;11:883-891. doi:10.1002/jbmr.5650110703.
  9. Krebsbach PH, Lee SK, Matsuki Y, Kozak CA, Yamada KM, Yamada Y. Full-length sequence, localization, and chromosomal mapping of ameloblastin. A novel toothspecific gene. J Biol Chem. 1996;271:4431-4435. https://doi.org/10.1074/jbc.271.8.4431
  10. Bartlett JD1, Simmer JP, Xue J, Margolis HC, Moreno EC. Molecular cloning and mRNA tissue distribution of a novel matrix metalloproteinase isolated from porcine enamel organ. Gene. 1996;183:123-128. https://doi.org/10.1016/S0378-1119(96)00525-2
  11. Hu JC1, Sun X, Zhang C, Liu S, Bartlett JD, Simmer JP. Enamelysin and kallikrein-4 mRNA expression in developing mouse molars. Eur J Oral Sci. 2002;110:307-315. https://doi.org/10.1034/j.1600-0722.2002.21301.x
  12. Iwasaki K, Bajenova E, Somogyi-Ganss E, Miller M, Nguyen V, Nourkeyhani H, Gao Y, Wendel M, Ganss B. Amelotin--a Novel Secreted, Ameloblast-specific Protein. J Dent Res. 2005;84:1127-1132. doi: 10.1177/154405910508401207.
  13. Moffatt P, Smith CE, St-Arnaud R, Nanci A. Characterization of Apin, a secreted protein highly expressed in toothassociated epithelia. J Cell Biochem. 2008;103:941-956. doi:10.1002/jcb.21465.
  14. Satoh T, Harada N, Hosoya T, Tohyama K, Yamamoto M, Itoh K. Keap1/Nrf2 system regulates neuronal survival as revealed through study of keap1 gene-knockout mice. Biochem Biophys Res Commun. 2009;380:298-302. doi:10.1016/j.bbrc.2009.01.063.
  15. Dinkova-Kostova AT, Holtzclaw WD, Kensler TW. The role of Keap1 in cellular protective responses. Chem Res Toxicol. 2005;18:1779-1791. doi: 10.1021/tx050217c.
  16. Kang MI, Kobayashi A, Wakabayashi N, Kim SG, Yamamoto M. Scaffolding of Keap1 to the actin cytoskeleton controls the function of Nrf2 as key regulator of cytoprotective phase 2 genes. Proc Natl Acad Sci U S A. 2004;101:2046-2051. 10.1073/pnas.0308347100.
  17. Yanagawa T1, Itoh K, Uwayama J, Shibata Y, Yamaguchi A, Sano T, Ishii T, Yoshida H, Yamamoto M. Nrf2 deficiency causes tooth decolourization due to iron transport disorder in enamel organ. Genes Cells. 2004;9:641-651. doi: 10.1111/j.1356-9597.2004.00753.x.
  18. Wakabayashi N, Itoh K, Wakabayashi J, Motohashi H, Noda S, Takahashi S, Imakado S, Kotsuji T, Otsuka F, Roop DR, Harada T, Engel JD, Yamamoto M. Keap1-null mutation leads to postnatal lethality due to constitutive Nrf2 activation. Nat Genet. 2003;35:238-245. doi: 10.1038/ng1248.
  19. Dassule HR1, Lewis P, Bei M, Maas R, McMahon AP. Sonic hedgehog regulates growth and morphogenesis of the tooth. Development. 2000;127:4775-4785.
  20. Haruyama N1, Thyagarajan T, Skobe Z, Wright JT, Septier D, Sreenath TL, Goldberg M, Kulkarni AB. Overexpression of transforming growth factor-beta1 in teeth results in detachment of ameloblasts and enamel defects. Eur J Oral Sci. 2006;114:30-34. doi: 10.1111/j.1600-0722.2006.00276.x.
  21. Wang XP, Suomalainen M, Jorgez CJ, Matzuk MM, Werner S, Thesleff I. Follistatin regulates enamel patterning in mouse incisors by asymmetrically inhibiting BMP signaling and ameloblast differentiation. Dev Cell. 2004;7:719-730. doi:10.1016/j.devcel.2004.09.012.
  22. Millar SE1, Koyama E, Reddy ST, Andl T, Gaddapara T, Piddington R, Gibson CW. Over- and ectopic expression of Wnt3 causes progressive loss of ameloblasts in postnatal mouse incisor teeth. Connect Tissue Res. 2003;44:124-129. https://doi.org/10.1080/03008200390152205
  23. Bouwman P1, Göllner H, Elsässer HP, Eckhoff G, Karis A, Grosveld F, Philipsen S, Suske G. Transcription factor Sp3 is essential for post-natal survival and late tooth development. EMBO J. 2000;19:655-661. doi: 10.1093/emboj/19.4.655.
  24. Nakamura T1, de Vega S, Fukumoto S, Jimenez L, Unda F, Yamada Y. Transcription factor epiprofin is essential for tooth morphogenesis by regulating epithelial cell fate and tooth number. J Biol Chem. 2008;283:4825-4833. DOI: 10.1074/jbc.M708388200.
  25. Bei M1, Stowell S, Maas R. Msx2 controls ameloblast terminal differentiation. Dev Dyn. 2004;231(4):758-65. doi:10.1002/dvdy.20182.
  26. Hinoi E, Takarada T, Fujimori S, Wang L, Iemata M, Uno K, Yoneda Y. Nuclear factor E2 p45-related factor 2 negatively regulates chondrogenesis. Bone. 2007;40:337-344. doi:10.1016/j.bone.2006.08.016.