| Project/Area Number |
06680638
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Functional biochemistry
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| Research Institution | TOKYO METROPOLITAN INSTITUTE FOR NEUROSCIENCE |
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Principal Investigator |
NAKATA Hiroyasu TOKYO METROPOLITAN INSTITUTE FOR NEUROSCIENCE,DERT.OF MOLECULAR & CELLULAR NEUROBIOLOGY,STAFF SCIENTIST, 神経生化学研究部門, 副参事研究員 (00041830)
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| Co-Investigator(Kenkyū-buntansha) |
SAITOH Yoshiko TOKYO METROPOLITAN INSTITUTE FOR NEUROSCIENCE,DERT.OF MOL.CELL.NEUROBIOL., STAFF, 神経生化学研究部門, 主事研究員 (70241263)
SAITOH Osamu TOKYO METROPOLITAN INSTITUTE FOR NEUROSCIENCE,DEPT.OF MOL.CELL.NEUROBIOL., STAFF, 神経生化学研究部門, 主事研究員 (60241262)
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| Project Period (FY) |
1994 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1995: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1994: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | Adenosine receptor / Purinergic / Cell culture / MAP kinase / Photoaffinity labeling / ATP / プリン受容体 / 培養細胞 / 細胞情報伝達 / ATP / Gタンパク / 情報伝達 / 細胞培養 / リン酸化 |
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| Research Abstract |
Studies on the signal transduction mechanism was performed using a cell culture system.We searched several cell lines which ecpress adenosine receptors.We found that cell lines such as DDT1MF-2 (syrian hamster smooth muscle-derived), ECV304 (human endothelial ) and N1E115 (mouse neuroblastoma) expressed A1 adenosine receptors.ECV304 and N1E115 also possessed another adenosine receptor subtype, A2b.By activation of A1 adenosine receptors by specific agonist, CPA,in these cell lines, it was found that mitogen-activated protein kinase (MAP kinase) which is known to play an essential role for the gene expression or cell proliferation was stimulated several fold.This activation was transient and retuned to the basal level in 30 min.Addition of several inhibitors for protein kinase C or PI-3-kinase inhibited the stimulation of MAP kinase, suggesting that protein kinase C and PI-3-kinase are involved in the MAP kinase cascade found in these cell lines described above.The similar agonist-induc
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ed activation of MAP kinase was observed in CHO cells which had been transfected with cDNA of rat A1 adenosine receptor.These results suggest that signals via adenosine receptors, especially A1 subtype, can control gene expression or cell proliferation in addition to the well-known cAMP system in cultured cells.We have also searched another subtype of adenosine receptors in order to explain many kinds of actions of adenosine in various cells or tissues.By using [3H] NECA binding as the marker, we could find a new adenosine binding protein in rat brain membranes which showed a unique ligand specificity, which is not classified into any known adenosine receptors.We developed a photoaffinity labeling system for this protein and determined the molecular mass (54 kDa) on SDS-PAGE.This protein was located in synaptic membranes.Purification of this protein is in progress to obtain partial amino acid sequence.Molecular cloning to determine its whole sequence and function will thus be preformed soon. Less
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