| Project/Area Number |
15591974
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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 |
Functional basic dentistry
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| Research Institution | Health Sciences University of Hokkaido |
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Principal Investigator |
ARAKAWA Toshiya Health Sciences University of Hokkaido, Dental School, Assistant Professor, 歯学部, 講師 (40306254)
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| Co-Investigator(Kenkyū-buntansha) |
TAKUMA Taishin Health Sciences University of Hokkaido, Dental School, Professor, 歯学部, 教授 (40095336)
HAKEDA Yoshiyuki Meikai Univ., Dental School, Professor, 歯学部, 教授 (90164772)
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| Project Period (FY) |
2003 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2004: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2003: ¥2,100,000 (Direct Cost: ¥2,100,000)
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| Keywords | mechanical stress / parathyroid hormon / osteoblast / PTH受容体 / 重力負荷 / 骨細胞 |
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| Research Abstract |
Parathyroid hormone (PTH) is a hormone that facilitate both osteogenesis and osteoclastic bone resorption. Osteogenesis was stimulated by intermittent administration of PTH, and bone resorption was atimulted repeated administration. However, its mechanism is unclear. We recently found that fluid shear stress, one of mechanical stress induced expression of mRNA of PTH receptor in MC3T3-Y4-A2 cells established from osteocytes in mouse long bone. In order to clarify the effects of PTH on osteogenesis, we constructed fusion protein with PTH receptor and Green fluorescence protein (GFP). First, we amplified a full length of cDNA of PTH receptor from cDNA that was synthesized from mRNA of MC3T3-Y4-A2 cells. Then, the full length of cDNA of PTH receptor was ligated into pEGFP-N1 and -C3 vectors (clontech co.). The EGFP vectors with PTH receptor cDNA were transfected into MC3T3-E1 oseoblast like cells established from mouse bone, and the cells stably expressing PTH receptor/GFP fusion protein were selected by neomysin. However, the cells that had high signals of fluorescence were not isolated because the cells that had bright signals were died. Though the reason was unclear, it assumed that excess signals of cAMP were transmitted in cells. The cells that had resistance to neomysin were finally isolated though the green fluorescence were not observed. The expression of mRNA of the fusion protein were identified by RT-PCR. We are examining the effects of mechanical stress on expression of PTH receptor with GFP.
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