Regulation of gene expression mediated by the opioid receptor and the nociceptin receptor

• Project/Area Number: 11307028
• Research Category: Grant-in-Aid for Scientific Research (A).
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Anesthesiology/Resuscitation studies
• Research Institution: KYOTO UNIVERSITY
• Principal Investigator: FUKUDA Kazuhiko Kyoto University Faculty of Medicine, Department of Anesthesia, Professor, 医学研究科, 教授 (90199224)
• Project Period (FY): 1999 – 2000
• Project Status: Completed (Fiscal Year 2000)
• Budget Amount: ¥33,300,000 (Direct Cost: ¥33,300,000); Fiscal Year 2000: ¥13,300,000 (Direct Cost: ¥13,300,000); Fiscal Year 1999: ¥20,000,000 (Direct Cost: ¥20,000,000)
• Keywords: Opioid / Transcription factor / Mitogen-activated protein kinase / Intravenous anesthetics / Ca^<2+> channel / Volatile anesthetics / K^+ channel / MAPモナーゼ / 耐性

Research Abstract

In this investigation, we mainly analyzed changes in gene expression induced by opioid analgesics and intravenous anesthetics, using molecular biological, biochemical and electrophysiological methods.
Activation of the opioid receptor expressed by transfection of the cloned cDNA in CHO cells induced expression of immediate early genes, c-fos and junB.This induction was inhibited by pertussis toxin and PD98059, suggesting that pertussis toxin-sensitive G-proteins and mitogen- avtivated protein kinase (MAPK) are involved in the signal transduction pathway. This mechanism might be involved in the molecular mechanism of opioid tolerance or dependence, serious side effects of opioid analgesics.
Effects of intravenous anesthetics on gene expression in neuronal cells have not been investigated. We found that midazolam induces expression of immediate early gene products, c-Fos and EGR-1, that was blocked by PD98059. Midazolam induced MAPK activation in a time- and dose-dependent manner. The midazolam-induced MAPK activation was blocked by inhibitors of tyrosine kinase and epidermal growth factor receptor. Our results may suggest that intravenous anesthetics induce long-term changes in neural functions by changes in gene expression patterns.
Chronic agonist exposure of the NG108-15 cells transformed to express the μ-opioid receptor was demonstrated to cause desensitization of the opioid-induced inhibition of the the N-type Ca^<2+> channel activity. The magnitude of the desensitization was different among various μ-opioid receptor-selective agonists.
Effects of volatile anesthetics on Ca^<2+>-activated K^+ channel subtypes were tested by the use of Xenopus oocyte expression system. It was demonstrated that halothane, isoflurane and sevoflurane do not affect the SK subtype, but suppressed the activity of the IK subtype. The physiological implication of this finding remains to be elucidated.

Research Products

• [Publications] Morikawa,H. et al.: "Opioid potentiation of N-type calcium channel currents via pertussis toxin-sensitive G proteins"Pflugers Arch.Eur.J.Physiol.. 438. 423-426 (1999)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Naoba,T. et al.: "Inhibition of the human intermediate conductance Ca^<2+>-activeted K^+ channel, hIKl, by volatile anesthetics"Eur.J.Pharmacol.. 395. 95-101 (2000)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Takeuchi,J. et al.: "Thioredoxin inhibite tumor neurosis factor-or interleukin-1-induced NF-kB activation at a level of Nf-kB-inducing kinase."Antioxidants & Redox Signaling. 2. 83-92 (2000)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Hirota,K. et al.: "Nucleoredoxin, glutaredoxin and thioredoxin differentially regulate NF-kB, AP-1, and CREB activation in HEK293 cells."Biochem. Biophys. Res. Commun.. 274. 177-184 (2000)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Shoda,T. et al.: "c-fos expression induced by activation of the u-opioid receptor in Chinese hamster ovary cells."Anesthesiology. 91. A745 (1999)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Shoda,T. et al.: "Phosphorylation of p90 ribosomel S6 kinase by mitogen-activated protein kinase in the signal transduction pathway of the μ opioid receptor"Anesthesiology. 93. A126 (2000)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Morikawa, H., Mima, H., Uga, H., Shoda, T.and Fukuda, K.: "Opioid potentiation of N-type calcium channel currents via pertussis toxin-sensitive G proteins."Pflugers Arch.Eur.J.Physiol. 438. 423-426 (1999)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Namba, T., Ishii, T.M., Ikeda, M., Hisano, T., Itoh, T., Hirota, K., Adelman, J.P.and Fukuda, K.: "Inhibition of the human Intermediate conductance Ca^<2+>-activated K^+ channel, hIK1, by volatile anesthetics."Eur.J.Pharmacol.. 395. 95-101 (2000)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Takeuchi, J., Hirota, K., Itoh, T., Shinkura, R., Kitada, K., Yodoi, J., Namba, T.and Fukuda, K.: "Thioredoxin inhibits tumor necrosis factor- or interleukin-1-induced NF-κB activation at a level upstream of NF-κB-inducing kinase."Antioxidants & Redox Signaling. 2. 83-92 (2000)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Hirota, K., Matsui, M., Murata, M., Takashima, Y., Cheng, F.S., Itoh, T., Fukuda, K.and Yodoi, J.: "Nucleoredoxin, glutaredoxin, and thioredoxin differentially regulate NF-kB, AP-1, and CREB activation in HEK293 cells."Biochem.Biophys.Res.Commun.. 274. 177-182 (2000)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Itoh, T., Hirota, K., Hisano, T., Namba, T.and Fukuda, K.: "Volatile anesthetics, halothane and isoflurane, differentially modulate proinflammatory cytokines-induced p38 mitogen activated protein kinase activation."Br.J.Anaesth.. (submitted.).
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Shoda, T., Fukuda, K., Uga, H., Mima, H.and Morikawa, H.: "Activation of μ-opioid receptor induces expression of c-fos and junB via mitogen-activated protein kinase cascade."Anesthesiology. (submitted).
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Fukuda, K., Shoda, T., Mima, H.and Uga, H.: "Midazolam-induced expression of c-Fos and EGR-1 in PC12 pheochromocytoma cell line."Anesthesiology. (submitted.).
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Shichino, T., Murakawa, M., Adachi, T., Miyazaki, Y., Segawa, H., Fukuda, K.and Mori, K.: "Effects of xenon on the release of acetylcholine in the rat cerebral cortex in vivo."Br.J.Anaesth. (submitted.).
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Shoda,T. et al.: "Phosphorylation of p90 rionsonal S6 kinase by initogen-actiurted protein kinase in the signal transduction pathway of the N opioid receptor."Anesthesiology. 93. A126 (2000)
• [Publications] Namba,T. et al.: "Inhibition of the human intermediate conductance Ca^<2+>-activited K^+ channel RIK1 by volatile cresthetics"Eur J Pharmacol. 395. 95-101 (2000)
• [Publications] Shoda,T.et al.: "c-fos expression induced by activation of the μ-opioid receptor in Chinese hamster ovary cells"Anesthesiology. Vol. 91, No. 3A. A745 (1999)