| Project/Area Number |
15390611
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Surgical dentistry
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| Research Institution | Mie University |
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Principal Investigator |
MURATA Taku Mie University, University Hospital, Research Associate, 医学部附属病院, 助手 (80242965)
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| Co-Investigator(Kenkyū-buntansha) |
TAGAWA Toshiro Mie University, Faculty of Medicine, Professor, 医学部, 教授 (30046346)
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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 |
¥13,000,000 (Direct Cost: ¥13,000,000)
Fiscal Year 2004: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 2003: ¥10,000,000 (Direct Cost: ¥10,000,000)
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| Keywords | osteoblast / phosphodiesterase 3 / phosphodiesterase 8 / phophodiesterase 3 / phophodiesterase 8 / phosphodiesterase |
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| Research Abstract |
Cyclic nucleotide phosphodiesterase(PDEs) catalyzes the hydrolysis of cAMP and cGMP, and are important in regulating intracellular concentrations and biological actions of these signal-transducing molecules. These enzymes contain at least 11 highly regulated and structurally related gene families (PDE1-11). However, little is known concerning the role of PDEs on bone formation. We studied the role of PDE8 on bone formation in mouse calvaria osteoblastic MC3T3-E1 cells, and bone structure of PDE3 knock-out mice.
Non-silencing controls, consisting of small interfering RNAs(siRNAs) that have no known homology with mammalian genes, are used to control for nonspecific silencing effects. The non-silencing control is labeled with Alex Fluor 488 and monitored transfection efficiency by fluoresce microscopy. As positive control mouse mitogen-activated protein kinases 1(MAPK1) is used and monitered gene silencing by reat-time PCR. Two mouse PDE8 siRNAs were synthesized and transfected in MC3T3-E1 cells. These PDE8 siRNAs changed bone maker mRNA expression, Runx2 and osteocalcin, in MC3T3-E1 cells.
Dr.Manganiello et al.in National Institute of Health(NIH) made PDE3 knock-out mice. The mice were viable and exhibited no obvious growth or developmental deficiencies.
These data suggested that PDE8 was important in bone formation, but not PDE3. We will try additional studies to assess the role of PDE8 and other PDEs on bone formation.
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