| Budget Amount *help |
¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000)
Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2013: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
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| Outline of Final Research Achievements |
We cloned HEK293 cells that constantly co-expressed HaloTag-tagged mu opioid receptors (MOR) and T7-tagged delta opioid receptors (DOR) (#25), and constantly co-expressed HaloTag-tagged DOR and FLAG-tagged MOR (#49). Signal transduction analysis using CellkeyTM system revealed that both #25 and #49 possessed MOR and DOR activity. #25 was treated with cell-impermeant HaloTag pH sensor ligand to follow the intracellular dynamics of hetero-dimerized mu-delta receptors that originally located on the membrane surface. DAMGO (a selective MOR agonist), SNC80 (a selective DOR agonist), morphine, fentanyl, and remifentanil induced internalization of heterodimerized MOR/DOR. Internalization induced by SNC80 showed that HaloTag-MOR/T7-DOR made a heterodimer. We are further evaluating the characteristics of intracellular dynamics of heterodimerized MOR/DOR under treatment of various concentrations of opioid agonists.
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