| Project/Area Number |
12671482
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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 |
Anesthesiology/Resuscitation studies
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| Research Institution | 宮崎医科大学 |
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Principal Investigator |
SUZUKI Nobuaki Miyazaki Medical College, Department of Anesthesia, Instructor, 医学部, 助手 (40206511)
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| Co-Investigator(Kenkyū-buntansha) |
NAKAMURA Tadashi Miyazaki Medical College, Department of Anesthesia, Instructor, 医学部, 助手 (60217859)
小佐井 和子 宮崎医科大学, 医学部, 助手 (00234740)
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| Project Period (FY) |
2000 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 2001: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2000: ¥2,100,000 (Direct Cost: ¥2,100,000)
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| Keywords | NMDA receptor / delta opioid receptor / CAMP / Mutant NMDA receptor / 変異型NMAD受容体遺伝子 / デルタ受容体 |
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| Research Abstract |
As a preliminary study, we planed to study the effects of mutant NMDA receptor expression on opioid tolerance of cultured cells.
N616R CDNA, 616th codon from N-terminal of NR1 cDNA, which is one of a NMDA subunit cDNA, has been mutated from AAC to CGC, was ligated into pcDNA3.1 vector. NR2A cDNA, which is a cDNA of another NMDA receptor subunit, was also ligated into pcDNA3.1 vector.
Neuroblastoma x glioma(NG108-15) hybrid cells were groum as monolayers in Dulbecco's modified Eagle's medium, containing 10 % fetal bovine serum. The cells were treated with control medium or medium containing delta-selective agonist (D-Pen2, D-Pen5)-enkephalin (DPDPE) for 4 to 12 hours. Thereafter, the cells were challenged with medium containing 10 μM forskolin and 500 μM 1-methyl-3-isobutylxanthine for 10 minutes. Production of cAMP was measured by enzymeimmunoassay. So far, DPDPE treatment did not cause forskolin stimulaeted hyper production of CAMP, which we planed to use as a measure for the opioid tolerance.
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