Identification of factors that modify either the heterodimerization of G protein-coupled receptors or the trafficking of heterodimerized receptors to the cell surface

• Project/Area Number: 15500262
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Neurochemistry/Neuropharmacology
• Research Institution: Nagasaki University
• Principal Investigator: UEZONO Yasuhito Nagasaki University, Graduate School of Biomedical Sciences, Associate Professor, 大学院・医歯薬学総合研究科, 助教授 (20213340)
• Co-Investigator(Kenkyū-buntansha): SHIBUYA Izumi University of Occupational and Environmental Health, School of Medicine, Associate Professor, 医学部, 助教授 (50162649) ; TANIYAMA Kohtaro Nagasaki University, Graduate School of Biomedical Sciences, Professor, 大学院・医歯薬学総合研究科, 教授 (70030898)
• Project Period (FY): 2003 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥3,800,000 (Direct Cost: ¥3,800,000); Fiscal Year 2004: ¥1,000,000 (Direct Cost: ¥1,000,000); Fiscal Year 2003: ¥2,800,000 (Direct Cost: ¥2,800,000)
• Keywords: GABA_B receptor / G protein-coupled receptors / Heterodimerization / Xenopus oocyte / FRET / Heterologous expression / Electrophyslology / Confocal Laser Microscopy / GABA-B受容体 / heterodimerization / heterolopous expression / 共焦点レーザー顕微鏡

Research Abstract

GABA is known to act as an inhibitory neurotransmitter on the central and peripheral nervous system. One of GABA receptors, the metabotropic GABAB receptor has been cloned in 1998. Functional GABA_B receptors are required to be formed with two GABAB receptor subunits, GABAB1 and GABAB2. Recent progress has also shown that not GABAB receptors but also many types of GPCRs such as μ- and δ-opioid receptors form heterodimer. Further, properties of such heterodimeric receptors have been reported to be differed from their parental monomeric receptors. However, nothing is known how heterodimers are formed and none of the compounds have been identified that modify the heterodimerization of GPCRs.
In the present study we focused on identification of factors that modify either formation of the heterodimerized GPCRs or trafficking of heterodimerized GPCR to the cell surface.
Our results showed that heterodimerization of GPCRs already occurred in the Golgi or endoplasmic reticulum and then heterodimer formed there were trafficked to cell surface. In addition we demonstrated that geldanamycin, an inhibitor of HSP90, which acts as chaperone in the cells, inhibited the trafficking of the heterodimerized receptors to the cells as well as formation of heterodimerization. These results indicated that some protein chaperone such as HSP 90 could be involved in the formation, and trafficking of the heterodimerized receptors to the cell surface.
We are now searching for compounds that modify formation and trafficking of the heterodimerized receptors. Such compounds will shed lights to open new category of drugs for modification of GPCR dimerization.

Research Products

• [Journal Article] Selective blockade of nicotinic acetylcholine reseptors by pimobendan, a drug for the treatment of heart failure : reduction of catecholamine secretion and synthesis in adrenal medullary cells. (2005)
  • Author(s): TOYOHIRA, Y., KUBO, T., UEZONO, Y.et al.
  • Journal Title: Naunyn-Schmiedeberg's Archives of Pharmacology 371 Pages: 107-113
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] 麻酔薬作用機序におけるG蛋白共役型受容体の関与 (2005)
  • Author(s): 南浩一郎, 上園保仁
  • Journal Title: 麻酔 54 Pages: 118-125
  • NAID: 40006628522
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Selective blockade of nicotinic acetylcholine receptors by pimobendan, a drug for the treatment of heart failure : reduction of catecholamine secretion and synthesis in adrenal medullary cells. (2005)
  • Author(s): TOYOHIRA, Y., KUBO, T., WATANABE, M., UEZONO, Y., YANAGIHARA, N., IZUMI, F.
  • Journal Title: Naunyn-Schmiedeberg's Archives of Pharmacology 371 Pages: 107-113
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] The effects of anesthetics on G protein-coupled receptors. (2005)
  • Author(s): Koichiro Minami, Yasuhito Uezono
  • Journal Title: Masui 54 Pages: 118-125
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] The effects of neurosteroids : pregnenoron, dehydroepiandrosterone, progesterone on muscarinic receptor-induced responses in Xenopus oocytes expressing cloned M1 and M3 receptors. (2005)
  • Author(s): NAKAMURA, M., MINAMI, K., UEZONO, Y., et al.
  • Journal Title: Naunyn Schmiedeberg's Archives of Pharmacology (In Press)
• [Journal Article] The effect of the tramadol metabolite O-demethyl tramadol, on the muscarinic receptor-induced responses in Xenopus oocytes expressing the cloned M1 or M3 receptors. (2005)
  • Author(s): NAKAMURA, M., MINAMI, K., UEZONO, Y., et al.
  • Journal Title: Anesthesia and Analgesia (In Press)
• [Journal Article] A forskolin derivative, colforsin daropate hydrochloride, inhibits the decrease in cortical renal blood flow induced by noradrenaline or angiotensin II in anesthetized nats. (2004)
  • Author(s): OGATA, J., MINAMI, K., UEZONO, Y.et al.
  • Journal Title: Nephron Physiology 96 Pages: 59-64
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Characterization of GABA_B receptors in the human colon. (2004)
  • Author(s): UEZONO Y., KAIBARA, M., KAWAKAMI, S et al.
  • Journal Title: Journal of Pharmacological Sciences 94 Pages: 211-213
  • NAID: 130000073876
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Inhibitory effects of tramadol on 5-hydroxytryptamine type 2C receptors expressed in Xenopus oocytes. (2004)
  • Author(s): OGATA, J., MINAMI, K., UEZONO, Y.et al.
  • Journal Title: Anesthesia and Analgesia 98 Pages: 1401-1406
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Characterization of GABA_B receptors involved in inhibition of motility associated with acetylcholine release in the dog small intestine : possible existence of a heterodimer of GABA_<B1> and GABA_<B2> receptors. (2004)
  • Author(s): KAWAKAMI, S., UEZONO, Y., MIKIMOTO, N.et al.
  • Journal Title: Journal of Pharmacological Sciences 94 Pages: 368-375
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Involvement of G βγ subunits in diverse signaling induced by G_<i/o> -coupled receptors : study using the Xenopus oocyte expression system. (2004)
  • Author(s): UEZONO, Y., KAIBARA, M., MURASAKI, O.et al.
  • Journal Title: Americal Journal of Physiology - Cell Physiology 287
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Analysis of the effects of halothane on G_i-coupled muscarinic M_2 receptor signaling in Xenopus oocytes using a chimeric Gα protein. (2004)
  • Author(s): MINAMI, K., UEZONO, Y., SHIRAISHI, M.et al.
  • Journal Title: Pharmacology 72 Pages: 205-212
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Inhibition of aortic smooth muscle cell proliferation by the intravenous anesthetic ketamine. (2004)
  • Author(s): SHIGA, Y., MINAMI, K., UEZONO, Y.et al.
  • Journal Title: Anesthesia and Analgesia 99 Pages: 1408-1412
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Characterization of functional effects of Z-338,a novel gastroprokinetic agent, on the muscarinic M_1,M_2 and M_3 receptors expressed in Xenopus oocytes. (2004)
  • Author(s): DOI, Y., MURASAKI, O., UEZONO, Y.et al.
  • Journal Title: European Journal of Pharmacology 505 Pages: 31-35
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] A forskolin derivative, colforsin daropate hydrochloride, inhibits the decrease in cortical renal blood flow induced by noradrenaline or angiotensin II in anesthetized rats. (2004)
  • Author(s): OGATA, J., MINAMI, K., SEGAWA, K., UEZONO, Y., SHIRAISHI M., YAMAMOTO, C., SATA, T., KIM, S-T., K.SHIGEMATSU, A.
  • Journal Title: Nephron Physiology 96 Pages: 59-94
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Characterization of GABA_B receptors in the human colon. (2004)
  • Author(s): UEZONO, Y., KAIBARA, M., KAWAKAMI, S., ENJOJI, A., TANIYAMA, K.
  • Journal Title: Journal of Pharmacological Sciences 94 Pages: 211-213
  • NAID: 130000073876
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Inhibitory effects of tramadol on 5-hydroxytryptamine type 2C receptors expressed in Xenopus oocytes. (2004)
  • Author(s): OGATA, J., MINAMI, K., UEZONO, Y., OKAMOTO, T., UETA, Y., SHIGEMATSU.A.
  • Journal Title: Anesthesia and Analgesia 98 Pages: 1401-1406
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Characterization of GABA_B receptors involved in inhibition of motility associated with acetylcholine release in the dog small intestine : possible existence of a heterodimer of GABA_<B1> and GABA_<B2> receptors. (2004)
  • Author(s): KAWAKAMI, S., UEZONO, Y., MIKIMOTO, N., ENJOJI, A., KAIBARA, M., KANEMATSU, T., TANIYAMA, K.
  • Journal Title: Journal of Pharmacological Sciences 94 Pages: 368-375
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Involvement of G βγ subunits in diverse signaling induced by G_<i/o>-coupled receptors : study using the Xenopus oocyte expression system. (2004)
  • Author(s): UEZONO, Y., KAIBARA, M., MURASAKI, O.TANIYAMA, K.
  • Journal Title: American Journal of Physiology-Cell Physiology 287
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Analysis of the effects of halothane on G_i-coupled muscarinic M_2 receptor signaling in Xenopus oocytes using a chimeric Gα protein. (2004)
  • Author(s): MINAMI, K., UEZONO, Y., SHIRAISHI, M., OKAMOTO, T., OGATA, J., TANIYAMA, K., SHIGEMATSU, A.
  • Journal Title: Pharmacology 72 Pages: 205-212
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Inhibition of aortic smooth muscle cell proliferation by the intravenous anesthetic ketamine. (2004)
  • Author(s): SHIGA, Y., MINAMI, K., SEGAWA, K., UEZONO, Y., SHIRAISHI, M., SATA T, Sung-Teh, K.
  • Journal Title: Anesthesia and Analgesia 99 Pages: 1408-1412
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Characterization of functional effects of Z-338, a novel gastroprokinetic agent, on the muscarinic M_1,M_2 and M_3 receptors expressed in Xenopus oocytes. (2004)
  • Author(s): DOI, Y., MURASAKI, O., KAIBARA, M., UEZONO, Y., HAYASHI, H., YANO, K., TAINYAMA, K.
  • Journal Title: European Journal of Pharmacology 505 Pages: 31-35
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Characterization of functional effects of Z-338, a novel gastroprokinetic agent, on the muscarinic M1, M2 and M3 receptors expressed in Xenopus oocytes. (2004)
  • Author(s): DOI, Y., MURASAKI, O., KAIBARA, M., UEZONO, Y., et al.
  • Journal Title: European Journal of Pharmacology 505 Pages: 31-35
• [Journal Article] Involvement of G βγsubunits in diverse signaling induced by Gi/o-coupled receptors : study using the Xenopus oocyte expression system. (2004)
  • Author(s): UEZONO, Y., KAIBARA, M., MURASAKI, O., TANIYAMA,K.
  • Journal Title: American Journal of Physiology-Cell Physiology 287
• [Journal Article] Analysis of the effects of anesthetics on the function of Gi-coupled muscarinic M2 receptors expressed in Xenopus oocytes using a chimeric Gα protein. (2004)
  • Author(s): MINAMI, K., UEZONO, Y., SHIRAISHI, M., et al.
  • Journal Title: Pharmacology 72 Pages: 205-212
• [Journal Article] Inhibition of aortic smooth muscle cell proliferation by the intravenous anesthetic ketamine. (2004)
  • Author(s): SHIGA, Y., MINAMI, K., SEGAWA, K., UEZONO, Y., et al.
  • Journal Title: Anesthesia and Analgesia 99 Pages: 1408-1412
• [Journal Article] Mechanism of cytosolic Ca^<2+> suppression by prostaglandin E_2 receptors in rat melanotrophs. (2003)
  • Author(s): NAGATA T, HARAYAMA N, SAKAKI N, INOUE M, TANAKA K, TOYOHIRA Y, UEZONO, Y et al.
  • Journal Title: Journal of Neuroendocrinology 15 Pages: 33-41
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Effects of the intravenously administrated anesthetics ketamine, propofol, and thiamylal on the cortical renal blood flow by in rats. (2003)
  • Author(s): SHIGA Y, MINAMI K, SEGAWA K, UEZONO Y et al.
  • Journal Title: Pharmacology 68 Pages: 17-23
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Destabilization of Na_v1.7 sodium channel a subunit mRNA by constitutive phosphorylation of ERK : negative regulation of steady-state level of cell surface functional sodium channels in adrenal chromaffin cells. (2003)
  • Author(s): YANAGITA T, KOBAYASHI H, UEZONO Y et al.
  • Journal Title: Molecular Pharmacology 63 Pages: 1125-1136
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Inhibition of the norepinephrine transporter function in cultured bovine adrenal medullary cells by Bisphenol A. (2003)
  • Author(s): TOYOHIRA Y, UTSUNOMIYA K, UEZONO Y.et al.
  • Journal Title: Biochemical Pharmacology 65 Pages: 2049-2054
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Inhibitory effects of ketamine and pentobarbital on substance P receptors expressed in Xenopus oocytes. (2003)
  • Author(s): OKAMOTO T., MINAMI K., UEZONO Y.et al.
  • Journal Title: Anesthesia and Analgesia 97 Pages: 104-110
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Inhibitory effects of alphaxalone on M_1 and M_3 muscarinic receptors expressed in Xenopus oocytes. (2003)
  • Author(s): SHIRAISHI, M., MINAMI, K., UEZONO, Y.et al.
  • Journal Title: Anesthesia and Analgesia 97 Pages: 449-455
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] 消化管運動とセロトニン (2003)
  • Author(s): 上園保仁, 貝原宗重, 林日出喜 他
  • Journal Title: Clinical Neuroscience 21 Pages: 656-657
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] 消化管運動における壁内神経系 (2003)
  • Author(s): 上園保仁, 貝原宗重, 林日出喜 他
  • Journal Title: 自律神経 40(2) Pages: 116-121
  • NAID: 50000263157
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] 受容体と食道機能 (2003)
  • Author(s): 谷山紘太郎, 貝原宗重, 上園保仁 他
  • Journal Title: 治療学 37(5) Pages: 27-30
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Involvement of 5-hydroxypriptamine (serotonin) receptors in intestinal motility and ion transport. (2003)
  • Author(s): TANIYAMA, K., SKURAI-YAMASHITA, Y., UEZONO, Y.et al.
  • Journal Title: The Journal of Surgical Metabolism and Nutrition 37 Pages: 171-178
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Mechanism of cytosoltc Ca^<2+> suppression by prostaglandin E_2 receptors in rat melanotrophs. (2003)
  • Author(s): NAGATA, T., HARAYAMA, N., SAKAKI, N., INOUE, M., TANAKA, K., TOYOHIRA, Y., UEZONO, Y., MARUYAMA, T., YANAGIHARA, N., UETA, Y., SHIBUYA, I.
  • Journal Title: Journal of Neuroendocrinology 15 Pages: 33-41
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Effects of the intravenously administrated anesthetics ketamine, propofol, and thiamylal on the cortical renal blood flow by in rats. (2003)
  • Author(s): SHIGA, Y., MINAMI, K., SEGAWA, K., UEZONO, Y., NAGAOKA, E., SHIRAISHI, M., NOGUCHI, T., SHIGEMATSU, A.
  • Journal Title: Pharmacology 68 Pages: 17-23
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Destabilization of Na_v1.7 sodium channel a subunit mRNA by constitutive phosphorylation of ERK : negative regulation of steady-state level of cell surface functional sodium channels in adrenal chromaffin cells. (2003)
  • Author(s): YANAGITA, T., KOBAYASHI, H., UEZONO, Y., YOKOO, H., SUGANO, T., SAITOH, T., MINAMI, S., SHIRAISHI, S., WADA, A.
  • Journal Title: Molecular Pharmacology 63 Pages: 1125-1136
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Inhibition of the norepinephrine transporter function in cultured bovine adrenal medullary cells by Bisphenol A. (2003)
  • Author(s): TOYOHIRA, Y., UTSUNOMIYA, K., UENO, S., MINAMI, K., UEZONO, Y., YOSHIMURA, R., TSUTSUI, M., IZUMI, F., YANAGIHARA, N.
  • Journal Title: Biochemical Pharmacology 65 Pages: 2049-2054
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Inhibitory effects of ketamine and pentobarbital on substance P receptors expressed in Xenopus oocytes. (2003)
  • Author(s): OKAMOTO, T., MINAMI, K., UEZONO, Y., OGATA, J., SHIGEMATSU, A., UETA, Y.
  • Journal Title: Anesthesia and Analgesia 97 Pages: 104-110
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Inhibitory effects of alphaxalone on M_1 and M_3 muscarinic receptors expressed in Xenopus oocytes. (2003)
  • Author(s): SHIRAISHI, M., MINAMI, K., SHIBUYA, I., OKAMOTO, T., UEZONO, Y., MURASAKI, O., KAIBARA, M., SHIGEMATSU, A.
  • Journal Title: Anesthesia and Analgesia 97 Pages: 449-455
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Serotonin and gastrointestinal motility. (2003)
  • Author(s): Yasuhito Uezono, Muneshige Kaibara, Hideki Hayashi, Kohtaro Taniyama
  • Journal Title: Clinical Neuroscience (In Japanese) 21 Pages: 656-657
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Peripheral autonomic nerves in digestive motility. (2003)
  • Author(s): Yasuhito Uezono, Muneshige Kaibara, Hideki Hayashi, Kohtaro Taniyama
  • Journal Title: Journal of the Japan Society of Neurovegetative Research 40(2) Pages: 116-121
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Functional esophageal movement and receptors. (2003)
  • Author(s): Kohtaro Taniyama, Muneshige Kaibara, Yasuhito Uezono, Hideki Hayashi
  • Journal Title: Biomedicine and Therapeutics 37(5) Pages: 27-30
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Involvement of 5-hydroxypriptamine (serotonin) receptors in intestinal motility and ion transport. (2003)
  • Author(s): TANIYAMA, K., SKURAI-YAMASHITA, Y., UEZONO, Y., KAIBARA, M., KANEMATSU T.
  • Journal Title: Journal of Surgical Metabolism and Nutrition 37 Pages: 171-178
  • Description: 「研究成果報告書概要(欧文)」より
• [Book] Current Topics in Drug Therapeutics (2005)
  • Author(s): MINAMI, K., UEZONO, Y.
  • Publisher: (In press)
• [Publications] MINAMI, K., UEZONO, Y., SHIRAISHI, M., OKAMOTO, et al.: "Analysis of the effects of anesthetics on the function of Gi-coupled muscarinic M2 receptors expressed in Xenopus oocytes using a chimeric Gα protein"Pharmacology. (In Press). (2004)
• [Publications] KAWAKAMI, S., UEZONO, Y., MIKIMOTO, N., ENJOJI, A., et al.: "Characterizarion of GABAB receptors involved in inhibition of motility associated with acetylcholine release in the dog small intestine"Journal of Pharmacological Sciences. (In Press). (2004)
• [Publications] UEZONO, Y., KAIBARA, M., KAWAKAMI, S., et al.: "Expression of GABAB receptors in the human colon"Journal of Pharmacological Sciences. 94(2). 211-213 (2004)
• [Publications] OGATA, J., MINAMI, K, SEGAWA, K., UEZONO, Y., et al.: "A forskolin derivative, colforsin daropate hydrochloride, inhibits the decrease in cortical renal blood flow induced by noradrenaline or angiotensin II in anesthetized rats"Nephron Physiol. 96(2). 59-64 (2004)
• [Publications] SHIRAISHI, M., MINAMI, K., SHIBUYA, I., OKAMOTO, T., UEZONO.Y., et al.: "Inhibitory effects of alphaxalone on M1 and M3 muscarinic receptors expressed in Xenopus oocytes"Anesthesia and Analgesia. 97. 449-455 (2003)
• [Publications] 上園保仁, 貝原宗重, 林日出喜, 谷山紘太郎: "消化管運動とセロトニン"Clinical Neuroscience. 21(6). 656-657 (2003)