Screening of Molecules for entrainment of circadian rhythms in mammalian central and peripheral clock cells

• Project/Area Number: 17310117
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: 基礎ゲノム科学
• Research Institution: Mitsubishi Kagaku Institute of Life Sciences
• Principal Investigator: TEI Hajime Mitsubishi Kagaku Institute of Life Sciences, Mitsubishi Kagaku Institute of Life Science, Research Group of Choronogenomics, Group Leader (00242115)
• Co-Investigator(Kenkyū-buntansha): SOGA Tomoyoshi Keio Univ., Faculty of Environment and Information Studies, Professor (60338217)
• Project Period (FY): 2005 – 2007
• Project Status: Completed (Fiscal Year 2007)
• Budget Amount: ¥14,560,000 (Direct Cost: ¥13,900,000、Indirect Cost: ¥660,000); Fiscal Year 2007: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000); Fiscal Year 2006: ¥3,700,000 (Direct Cost: ¥3,700,000); Fiscal Year 2005: ¥8,000,000 (Direct Cost: ¥8,000,000)
• Keywords: Circadian rhythms / Suprachiasmatic nucleus / Clock gene / Metabolome / Capillary electrophoresis-Mass spectrometry(CE-MS) / 視交叉上刻 / データベース / 視交叉上核

Research Abstract

Many biochemical, physiological and behavioral processes in many organisms exhibit circadian rhythms. The central circadian oscillator of mammals is located at the suprachiasmatic nucleus(SCN) of the brain. The circadian fluctuation of electrophysiological activities and/or humoral factors produced by the SCN regulates the peripheral rhythms. The mechanism for the generating carcadian rhythms is regarded as a network structure involving complicated molecular interactions. Thus, the elucidation of the structure of this molecular network requires modeling methods based on an integrated database constructed with the comprehensive quantafcation of transcripts, proteins, and metabolite in the master clock cells. In this study, we first established a rat SCN derived cell line from a transgenic rat in which the mouse Per1 gene promoter is linked to a luciferase reporter. Light emission from the SCN derived cell line was robustly rhythmic in static culture. The time dependent phase advance or delay of the circadian rhythm was accomplished after the treatment of the cell line with forskolin. In addition, we have established an efficient metabolome system to estimate cellular metabolites utilizing an extremely sensitive Capillary electrophoresis-Mass spectrometry(CE-MS) technique. Then we analyzed approximately 800 metabolites in the SCN derived cell line using these methods, and constructed a molecular database that collected metabolites with a circadian fluctuation and a rapid increase after forskolin treatment in the cellular level. A prominent oscillation of the markers of the cellular energy charge, ATP, ADP, NADP, and NADPH, indicated that the energy metabolism was under the control of circadian rhythms. In addition, the energy charge after phase advance felt into an extremely low level in comparison with that after phase delay. The results reflected a fact that a longer transition time was necesary for phase advance than that for phase delay.

Research Products

• [Journal Article] LARK activates post-transcriptional expression of an essential mammalian clock protein, PERIOD1. (2007)
  • Author(s): Kojima, S., Matsumoto, K., Hirose, M., Shimada, M., Nagano, M., Shigeyoshi, Y., Hoshino, S., Green, C.B., Sakaki, Y., and Tei, H.
  • Journal Title: Proc. Natl. Acad. Sci. USA 104 Pages: 1859-1864
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Establishment of cell lines derived from the rat suprachiasm atic nucleus. (2007)
  • Author(s): Kawaguchi, S., Shinozaki, A., Obinata, M., Saigo, K., Sakaki, Y., Tei, H.
  • Journal Title: Biochem. Biophys. Res. Commun. 355 Pages: 555-561
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Bimodal clock gene expression in mouse suprachiasmatic nucleus and peripheral tissues under a 7-hour light and 5-hour dark schedule. (2007)
  • Author(s): Watanabe, T., Naito, E, Nakao.N, Tei, H., Yoshimura, T., Ebihara, S.
  • Journal Title: J. Biol. Rhythms. 22 Pages: 58-68
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Roles of Hemoglobin Allostery in Hypoxia-induced Metabolic Alterations in Erythrocytes: Simulation and Its Demonstration by Metabolome Analysis (2007)
  • Author(s): Kinoshita, A., Tsukada, K., Soga, T., Hishiki, T., Ueno, Y., Nakayama, Y., Tomita, M., Suematsu, M.
  • Journal Title: J. Biol. Chem. 282 Pages: 10731-10741
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] LARK activates post-transcriptional expression of an essential mammalian clock protein, PERIOD1. (2007)
  • Author(s): Kojima, S., Matsumoto, K., Hirose, M., Shimada, M., Nagano, M., Shigeyoshi, Y., Hoshino, S., Green, C. B., Sakaki, Y., and Tei, H.
  • Journal Title: Proc. Natl. Acad. Sci. USA 104 Pages: 1859-1864
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Establishment of cell lines derived from the rat suprachiasmatic nucleus. (2007)
  • Author(s): Kawaguchi, S., Shinozaki, A., Obinata, M., Saigo, K., Sakaki, Y., Tei, H.
  • Journal Title: Biochem. Biophys. Res. Commun. 355 Pages: 555-561
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Bimodal clock gene expression in mouse suprachiasmatic nucleus and peripheral tissues under a 7-hour light and 5-hour dark schedule. (2007)
  • Author(s): Watanabe, T., Naito, E, Nakao, N., Tei, H., Yoshimura, T., Ebihara, S.
  • Journal Title: J. Biol. Rhythms. 22 Pages: 58-68
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Roles of Hemoglobin Allostery ui Hypoxia-induced Metabolic Alterations in Erythrocytes : Simulation and Its Demonstration by Metabolome Analysis (2007)
  • Author(s): Kinoshita, A., Tsukada, K., Soga, T., Hishiki, T., Ueno, Y., Nakayama, Y., Tomita, M., Suematsu, M.
  • Journal Title: J. Biol. Chem. 282 Pages: 10731-10741
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Bimodal clock gene expression in mouse suprachiasmatic nucleus and peripheral tissues under a 7-hour light and 5-hour dark schedule. (2007)
  • Author(s): Watanabe, T., Naito, E, Nakao. N, Tei, H., Yoshimura, T., Ebihara, S.
  • Journal Title: J. Biol. Rhythms. 22 Pages: 58-68
  • Peer Reviewed
• [Journal Article] LARK activates post-transcriptional expression of an essential mammalian clock protein, PERIOD1. (2007)
  • Author(s): Kojima, S., Matsumoto, K., Hirose, M., Shimada, M., Nagano, M., Shigeyoshi, Y., Hoshino, S., Green, C.B., Sakaki, Y., Tei, H.
  • Journal Title: Proc. Natl. Acad. Sci. USA 104 Pages: 1859-1864
• [Journal Article] Multiple Highthroughput Analyses Monitor The Response of E. coli to Perturbations (2007)
  • Author(s): Ishii, N., Nakahigashi, K., Baba, T., Robert, M., Soga, T., Kanai, A., Hirasawa, T., et al.
  • Journal Title: Science (In press)
• [Journal Article] Roles of Hemoglobin Allostery in Hypoxia-induced Metabolic Alterations in Erythrocytes : Simulation and Its Demonstration by Metabolome Analysis (2007)
  • Author(s): Kinoshita, A., Tsukada, K., Soga, T., Hishiki, T., Ueno, Y., Nakayama, Y., Tomita, M., Suematsu, M.
  • Journal Title: J. Biol. Chem. 282 Pages: 10731-10741
• [Journal Article] Constitutive Expression of the Period1 Gene Impairs Behavioral and Molecular Circadian Rhythms. (2006)
  • Author(s): Numano, R., Yamazaki, S., Umeda, N., Samura, T., Sujino, M., Takahashi, R., Ueda., M., Mori, A., Yamada, K., Sasaki, Y., Inoue, S., Menaker, M., Tei, H.
  • Journal Title: Proc. Natl. Acad. Sci. USA 103 Pages: 3716-3721
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] MathDAMP: a package for differential analysis of metabolite profiles (2006)
  • Author(s): Baran, R., Kochi, H., Saito, N., Suematsu, M., Soga, T., Nishioka, T., Robert, M., Tomita, M.
  • Journal Title: BMC Bioinformatics 7 Pages: 530-530
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] A Metabolomics Approach for Enzyme Discovery (2006)
  • Author(s): Saito, N., Robert, M., Kitamura, S., Baran, R., Soga, T., Mori, H., Nishioka, T., Tomita, M.
  • Journal Title: J. Proteome Res. 5 Pages: 1979-1987
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Constitutive Expression of the Period1 Gene Impairs Behavioral and Molecular Circadian Rhythms. (2006)
  • Author(s): Numano, R., Yamazaki, S., Umeda, N., Samura, T., Sujino, M., Takahashi, R., Ueda, M., Mori, A., Yamada, K., Sasaki, Y., Inoue, S., Menaker, M., Tei, H.
  • Journal Title: Proc. Natl. Acad. Sci. USA 103 Pages: 3716-3721
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] MathDAMP : a package for differential analysis of metabolite profiles (2006)
  • Author(s): Baran, R., Kochi, H., Saito, N., Suematsu, M., Soga, T., Nishioka, T., Robert., M., Tomita, M.
  • Journal Title: BMC Bioinformatics 7 Pages: 530-530
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] A Metabolomics Approach for Enzyme Discovery (2006)
  • Author(s): Saito, N., Robert., M., Kitamura, S., Baran, R., Soga, T., Mori, H., Nishioka, T., Tomita, M.
  • Journal Title: J. Proteome Res. 5 Pages: 1979-1987
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Constitutive Expression of the Period1 Gene Impairs Behavioral and Molecular Circadian Phythms. (2006)
  • Author(s): Numano, R., Yamazaki, S., Umeda, N., Samura, T., Sujino, M., Takahashi, R., Ueda, M., Mori, A., Yamada, K., Sakaki, Y., Inouye, S., Menaker, M., Tei, H.
  • Journal Title: Proc. Natl. Acad. Sci. USA 103 Pages: 3716-3721
• [Journal Article] MathDAMP : a package for differential analysis of metabolite profiles (2006)
  • Author(s): Baran, R., Kochi, H., Saito, N., Suematsu, M., Soga, T., Nishioka, T., Robert, M., Tomita, M.
  • Journal Title: BNC Bioinformatics 7 Pages: 530-530
• [Journal Article] Constitutive Expression of the Period1 Gene Impairs Behavioral and Molecular Circadian Rhythms. (2006)
  • Author(s): Numano, R., Yamazaki, S., Umeda, N., Samura, T., Sujino, M., Takahashi, R., Ueda, M., Mori, A., Yamada, K., Sakaki, Y., Inouye.S., Menaker, M., Tei, H.
  • Journal Title: Proc. Natl. Acad. Sci. USA 103 Pages: 3716-3721
• [Journal Article] A Calcium Flux is Required for Circadian Rhythm Generation in Mammalian Pacemaker Neurons (2005)
  • Author(s): Lundkvist, G.B., Kwak, Y., Davis, E.K., Tei, H., Block, G.D.
  • Journal Title: J. Neuroscience 25 Pages: 7682-7686
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Metabolome Analysis and Metabolic Simulation (2005)
  • Author(s): Ishii, N., Soga, T., Nishioka, T., Tomita, M.
  • Journal Title: Metabolomics 1 Pages: 29-37
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Large-Scale Prediction of Cationic Metabolites Identity and Migration Time in Capillary Electrophoresis Mass Spectrometry Using Artificial Neural Networks (2005)
  • Author(s): Sugimoto, M., Kikuchi, S., Arita, M., Soga, T., Nishioka, T., Tomita, M.
  • Journal Title: Anal. Chem. 77 Pages: 78-84
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] A Calcium Flux is Required for Circadian Rhythm Generation in Mammalian Pacemaker Neurons (2005)
  • Author(s): Lundkvist., G. B., Kwak., Y., Davis., E. K., Tei, H., Block., G. D.
  • Journal Title: J. Neuroscience 25 Pages: 7682-7686
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] A Calcium Flux is Required for Circadian Rhythm Generation in Mammalian Pacemaker Neurons. (2005)
  • Author(s): Lundkvist, G.B., Kwak, Y., Davis, E.K., Tei, H., Block, G.D.
  • Journal Title: J.Neuroscience 25 Pages: 7682-7686
• [Journal Article] Metabolome Analysis and Metabolic Simulation. (2005)
  • Author(s): Ishii, N., Soga, T., Nishioka, T., Tomita, M.
  • Journal Title: Metabolomics 1 Pages: 29-37
• [Journal Article] Large-Scale Prediction of Cationic Metabolites Identity and Migration Time in Capillary Electrophoresis Mass Spectrometry using Artificial Neural Networks. (2005)
  • Author(s): Sugimoto, M., Kikuchi, S., Arita Tomita, M.
  • Journal Title: Anal. Chem. 77 Pages: 78-84