| Project/Area Number |
15590257
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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 |
General medical chemistry
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| Research Institution | Kansai Medical University |
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Principal Investigator |
OKUDA Emiko Kansai Medical University, Faculty of Medicine, Assistant Professor, 医学部, 講師 (50291802)
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| Co-Investigator(Kenkyū-buntansha) |
ITO Seiji Kansai Medical University, Faculty of Medicine, Professor, 医学部, 教授 (80201325)
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| Project Period (FY) |
2003 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2004: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 2003: ¥1,900,000 (Direct Cost: ¥1,900,000)
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| Keywords | Nocistatin / Nociceptin / orphanin FQ / Pain transmission / BRET / Processing / Proprotein convertase / Scretion pathway / Prostaglandin E_2 |
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| Research Abstract |
Nocistatin (NST) and nociceptin/orphanin FQ (N/OFQ) are derived from the same precursor protein, while NST exhibits antagonism against N/OFQ-induced pain responses. The regulation of formation and release of these peptides is very important for the opposite pain responses. We developed an intra-molecular BRET system for monitoring dynamic biological process of the production of NST and N/OFQ in living cells. To clarify the regulation of pain responses between NST and N/OFQ, we carried out the identification of regulated molecules for the production and release of these peptides using the processing monitoring BRET probe in living cells. Furthermore, we analyzed the pain transmission in relation to the release of N/OFQ and NST in vivo.
1.Formation and release of NST and N/OFQ : Processing enzymes. proprotein convertase (PC) 1,PC2, and furin were involved in the formation of NST and N/OFQ in living cells using the processing monitoring BRET probe. NST was released through the constitutive secretion pathway in PC1 and furin-expressing cells, while it was released through the regulated secretion pathway in PC2 and furin-expressing cells. Expression of PC2 and furin mRNA increased in the spinal cord of i.pl. complete Freund's adjuvant-injected mice. These results suggested that the increase of the processing enzyme mRNAs induced the difference of formation and release of NST, resulting in regulation of N/OFQ-evoked pain responses.
2.Pain transmission via the release of NST and N/OFQ : Prostaglandin (PG) E_2 induced allodynia by stimulation of N/OFQ release in the spinal cord. The administration of i.t. NST inhibited the PGE_2- induced allodynia. The release of NST and N/OFQ was involved in the regulation of pain transmission.
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