2002 Fiscal Year Final Research Report Summary
STUDY ON PROSTAGLANDIN F2α-MEDIATED SIGNAL TRANDUCTION MECHANISM IN THE OSTEOBLAST
| Project/Area Number |
12671931
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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 |
Surgical dentistry
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| Research Institution | GIFU UNIVERSITY |
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Principal Investigator |
KATO Yukihiro GIFU UNIVERSITY, SCHOOL OF MEDICINE, DEPARTMENT OF ORAL AND MAXILLOFACIAL SURGERY, STAFF ORAL AND MAXILLOFACIAL SURGEON, 医学部, 助手 (30293567)
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| Co-Investigator(Kenkyū-buntansha) |
TOIDA Makoto GIFU UNIVERSITY, SCHOOL OF MEDICINE, DEPARTMENT OF ORAL AND MAXILLOFACIAL SURGERY, ASSOCIATE PROFESSOR, 医学部, 助教授 (90313890)
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| Project Period (FY) |
2000 – 2002
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| Keywords | Osteoblast-like cells / Signaltransduction / Prostaglandin F2α / Phospholipase D / GTP-binding protein |
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| Research Abstract |
Prostaglandin F2α (PGF2α) has been considered as one of the cell growth factors. The biochemical signal transduction mechanism in osteoblasts stimulated by PGF2α still remain unclear. In order to know PGF2α-receptor signaling to Phospholipase D (PLD), we have investigated the role of GTP-binding protein in PGF2α-mediated PLD activation in the osteoblast-like cell line MC3T3-E1
PLD activity was determined by measuring [^3H]Phosphatidylbuthanol ([^3H]PtdBut) produced via PLD-catalyzed transphosphatidylation in [^3H]myristic acid-labeled cells
1. In intact cells, cells were stimulated by 1μM of PGF2α for 10 min. in the presence of 0.25%Butanol, [^3H]PtdBut generation increased 7-fold (1.32±0.05%) compared with control cells (0.08±0.03%)
2. To examine the role of the receptor-coupled GTP-binding protein on PLD activation stimulated by PGF2α, cells were stimulated by Guanosine 5'-O-(3-thiotriphosphate) (GTPγS) in the presence of 0.25%Butanol in digitonin-permeabilized cells, [^3H]PtdBut accumulation was observed in a dose-dependent manner (10,μM〜100μM) with a maximum at 20μM, and was greatly increased by GTPγS (20 : RM) in a time-dependent manner (〜60min.). At 30min. of incubation, [^3H]PtdBut generation stimulated by GTP γS increased 16-fold (4.16±0.50%) compared with control cells (0.26±0.30%), respectively. Small increases were observed in the presence of Ca^<2+> (1μM) and Mg-ATP (1 mM). Both Ca^<2+> and Mg-ATP were required for full activation of GTPγS-stimulated PLD in the osteoblast-like cells
3. No effect was observed in the GTPγS-stimulated [^3H]PtdBut formation in the permeabilized cells which were pretreated with pertussis toxin (100 ng/ml) for 12h, suggesting that Gq and other type(s) of GTP-binding protein were involved in PGF2a -dependent PLD activation
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