| Project/Area Number |
63480412
|
|---|
| Research Category |
Grant-in-Aid for General Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
Functional basic dentistry
|
|---|
| Research Institution | Dental School Okayama University |
|---|
Principal Investigator |
TANIGUCHI Shigehiko Okayama Univ.Dent.Sch. Professor, 歯学部, 教授 (50034161)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
NISHI Mieko Okayama Univ.Dent.Sch. Assistant, 歯学部, 教務員 (40180575)
TAKAHASHI Kojiro Okayama Univ.Dent.Sch. Instructor, 歯学部, 助手 (00144775)
KAWAMOTO Tomoyuki Okayama Univ.Dent.Sch. Lecturer, 歯学部, 非常勤講師
川本 奉之 岡山大学, 歯学部, 非常勤講師
|
|---|
| Project Period (FY) |
1988 – 1989
|
| Project Status |
Completed (Fiscal Year 1989)
|
| Budget Amount *help |
¥5,000,000 (Direct Cost: ¥5,000,000)
Fiscal Year 1989: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1988: ¥4,600,000 (Direct Cost: ¥4,600,000)
|
|---|
| Keywords | Matrix vesicle / Alkaline phosphatase / Acid phosphatase / Osteoblast-like cell / MC3T3-EI / アルカリホスファターゼ / 酸性ホスファターゼ / MC3T3ーEI |
|---|
| Research Abstract |
The molecular process leading to apatite formation in matrix vesicle(MV) during the initial phase of calcification has never been understood so far on the basis of functional behavior of the intravesicular phosphatases. The project has been promoted with two types of MV participating each in different modes of calcification.
(1) MV from enchondral ossification process: The MV from calf epiphyseal cartilage fractionated finally through Percoll density gradient centrifugation showed the highest alkaline phosphatase(ALP) specific activity among other organelles. The phosphotransferase activity of the ALP is interpreted as an essential function in calcification in or by MV. Apparent heterogeneity of MV suggested from the unfixed ratio of acid phosphatase(ACP)/ALP activity did not permit to ascribe the ACP as a unique vesicular enzyme despite possible localization of non-lyzosomal ACP in MV.
(2) MV from intramembranous ossification process: The cultured cell of osteoblastic clone MC3T3-EI from mouse calvaria was examined in respect of the MV formation. In-between the ending of the monolayer growth where ALP level elevates in consequence of the retinoic acid receptor genes expression and the starting of the multi-layer growth together with Ca deposit, has been shown to be critical timing for the MV release. Thus, the criteria for sampling of calcifying MV have been established.
(3) Automated micro stopped flow apparatus for rapid phosphatase assay: The apparatus constructed was powerful in dealing with very limited amounts of MV preparation in case of the fractionation or kinetic assay. Taking advantage of the above new apparatus, the MV from the osteoblastlike cultured cells is proved to be fruitful experimental system.
|
