| Project/Area Number |
18590169
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
General anatomy (including Histology/Embryology)
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| Research Institution | Kyushu University |
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Principal Investigator |
KOBAYASHI Ieyoshi Kyushu University, Faculty of Dental Science, Associate Professor (40243951)
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| Co-Investigator(Kenkyū-buntansha) |
SAKAI Hidetaka Kyushu University, Faculty of Dental Science, Professor (80136499)
KIYOSHIMA Tamotsu Kyushu University, Faculty of Dental Science, Assistant Professor (20264054)
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| Project Period (FY) |
2006 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥3,920,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥420,000)
Fiscal Year 2007: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2006: ¥2,100,000 (Direct Cost: ¥2,100,000)
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| Keywords | embyologir / morphology / tooth germ / knockdown method / thvmosin beta 4 / nucleolin / anti-sense nucleic acid method / Runx2 / Thymosin beta 4 |
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| Research Abstract |
Odontogenesis is a complex developmental process that is mediated by a series of epithelial-mesenchymal interactions. We previously detected several intensely expressed genes, e.g. phosphoglycerate kinase 1(Pgk 1), Nucleolin, thymosin beta 4 (Tβ4) and so on at the initial stages of tooth development. Tβ4 was found to be differentially expressed in embryonic day 12 (E12) mouse mandible. Tβ4 was expressed in the dental epithelium at E10.5-18. At E18. In this research project, the function of Tβ4 in developing tooth germ was checked by using Tβ4 antisense phosphorothioated oligonucleotide (AS-S-ODN) in cultured E11 mouse mandible and E 15 tooth germ. Complete suspension of tooth germ formation was observed after 8 days culture of E11 mandible. Inhibition of differentiation of mesenchymal cells to odontoblasts cells was observed after 8 days culture of E15 tooth germ. Deposition of enamel and dentin matrix was also inhibited in the tooth germs cultured with AS-S-ODN. Semi quantitative RT-P
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CR was performed to confirm the inhibition of expression of Tβ4 mRNA, and also to check the effect of Tβ4 AS-ODN on the expression of matrix metalloproteinase (MMP) -2/-9, dentin sialophosphoprotein, dentin marix protein 1, amelogenin and enamelin genes in odontogenic cell culture. Inhibition of Tβ4, MMP-2/-9 and odontogenic factors were detected in Tβ4 AS-S-ODN treated cells. This inhibition assay demonstrates that Tβ4 has important role in morphogenesis of tooth germ and in dentin matrix formation. This result represents that Tβ4 may play role in odontogenesis by controlling expressions of MMPs and odontogenic factors.
In an inhibition assay for Nucleolin, Nucleolin AS S-ODN treatment resulted in an arrest of tooth germ growth at the cap-like stage in cultured E15 mandible. This inhibition assay indicated that Nucleolin took part in the tooth germ initiation and morphogenesis during early stage of tooth germ development. The strong expression of Pgkl mRNA was also seen in theodontogenic epithelial cells and surroundingmesenchymal cells of the tooth germ at E10.5-18.0. Pgkl protein formed 84-kDa protein complex in the embryonic organs. These results suggested this complex to be formed with glyceraldehyde-3-phosphate dehydrogenase(GAPDH), and Pgklplays some functional roles in the development of tooth germ by forming protein complex with GAPDH. We also reported Nucleolin and Pgkl functional roles in the development of tooth in journals (Journal of Biological Chemistry & Histology and Histopathology). Less
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