| Project/Area Number |
25640024
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Nerve anatomy/Neuropathology
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| Research Institution | Kyoto Prefectural University of Medicine |
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Principal Investigator |
TANAKA Masaki 京都府立医科大学, 医学(系)研究科(研究院), 准教授 (80264753)
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| Co-Investigator(Kenkyū-buntansha) |
WATANABE Yoshihisa 京都府立医科大学, 大学院医学研究科, 講師 (50363990)
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| Project Period (FY) |
2013-04-01 – 2015-03-31
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| Project Status |
Completed (Fiscal Year 2014)
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| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2013: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
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| Keywords | 5-HT2C受容体 / Tango / GPCR / arrestin / RNA編集 / Arrestin / スクリーニング系 / 5-HT2CR |
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| Outline of Final Research Achievements |
In this study, we developed a serotonin (5-HT) 2C receptor (5-HT2CR)-Tango assay system, a novel analysis tool of 5-HT2CR activity based on the G protein coupled receptor-arrestin interaction. In the first year, we constructed pEF1-5-HT2CR-Tango plasmids by modifying plasmids for Tango-system in drosophila with insertion of 5-HT2CR gene. We used two isoforms of mRNA editing, that is, INI and VGV isoforms of 5-HT2CR. Then we constructed plasmids for EGFP-reporter whose expression reflects the amount of released LexA by arrestin. In the second year, we developed a cell line stably expressing 5-HT2CR-Tango system using HEK293 cells and measured activity of INI and VGV isoforms and confirmed this system worked well. We think this system will be applied for high throughput drug-screening of 5-HT2CR activating ligands and in vivo brain imaging of 5-HT2CR activity after developing 5-HT2CR-Tango-transgenic mice in the future.
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