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Real time monitoring of mammalian circadian rhythm by bio-sensor

Research Project

Project/Area Number 08557007
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section展開研究
Research Field Environmental physiology (including Physical medicine and Nutritional physiology)
Research InstitutionKobe University

Principal Investigator

OKAMURA Hitoshi  Kobe University, Department of Anatomy and Brain Science, Professor, 医学部, 教授 (60158813)

Co-Investigator(Kenkyū-buntansha) T.INOUYE Shin-Ichi  Yamaguchi University, Faculty of Science, Professor, 理学部, 教授 (10274151)
SHINOHARA Hiroaki  Okayama University, Department of Bioscience and Biotechnology, Faculty of Engin, 工学部, 助教授 (60178887)
Project Period (FY) 1996 – 1998
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥7,600,000 (Direct Cost: ¥7,600,000)
Fiscal Year 1998: ¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 1997: ¥4,000,000 (Direct Cost: ¥4,000,000)
Keywordsclock genes / luciferase- / mPer1-promotor / bioaffinity binding / ルシフェラーゼ / 時計遺伝子 / サーカディアンリズム / フォトン検出装置 / Rat1 / 視交叉上核 / ピリオド遺伝子 / 概日リズム
Research Abstract

It is considered that protein assembly on solid substrates is the important key technology to fabricate high performance biosensing systems and bioelectronics devices. The purpose of this study is to develop novel protein assembling method using bioaffinity binding and dissociation. The author paid attention to bioaffinity binding reaction between biotin analogues and avidin molecule. It is expected that biotin analogue-modified protein molecules are assembled on the solid supports by using avidin molecules as binders and that the constructed protein complexes are decomposed by the exchange reaction with free biotin molecules. This assembling technique might be available to fabricate recycle usable biosensing system and intelligent bioreactor system in which biocatalysis is controlled by the addition of ligand molecules as stimulation signal.
Bioaffinity binding and dissociation reactions of avidin and biotin- or biotin analogue-modified proteins were characterized with evanescent wave- … More exciting fluorescence measurement. Dissociation of avidin and biotin analogue-modified protein complexes was certainly induced by the free biotin addition. It was demonstrated that biotin sensing is possible in the concentration range from 1x10^<-9> M to 1x10^<-5> M by the measurement of dissociation degree. The data further suggested that the biotin concentration for dissociation induction is dependent on the bioaffinity between the biotin analogue and avidin.
At second, It was demonstrated that redox coenzyme-modified avidin molecules were assembled on the biotin-modified electrode surface and functioned as bioelecfrocatalyst for NADH oxidation. Dissociation of themodified avidin from the biotin-modified electrode surface was also induced by biotin addition.
During three years we cloned putative mammalian clock genes (mPer1, mPer2, mPer3 and mTim). mPer1-promotor-Luciferasefusion gene was transfected to the Rat1 fibroblast cells. We observed the change of luminescence at single cell level by a sensitive photon detector. After serum shock, the luciferin luminescence begins to show the rhythm in a period length between 20 to 28 hours. Less

Report

(4 results)
  • 1998 Annual Research Report   Final Research Report Summary
  • 1997 Annual Research Report
  • 1996 Annual Research Report
  • Research Products

    (38 results)

All Other

All Publications (38 results)

  • [Publications] Tei,H.: "Circadian oscillation of a mammalian homologue of the Drosophila period gene." Nature,. 389,. 512-516. (1997.)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Shigeyoshi,Y.: "Light-induced resetting of a mammalian circadian clock is associated with rapid induction of the mPer1 transcript." Cell,. 91. 1043-1053, (1997.)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Ban Y.: "Development of circadian VIP rhythm in the rat suprachiasmatic nucleus." J.Neurosci.,. 17,. 3920-3931, (1997.)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Shinohara,K.: "Luminance-dependent decrease in vasoactive intestinal polypeptide in the suprachiasmatic nucleus." Neurosci. Lett.251巻. 21-24 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Takumi T.: "A light independent oscillatory gene mPer3 in mouse SCN and OVLT." EMBO J.17巻. 4753-4759 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] 篠原寛明: "水晶振動子マイクロバランスを用いる動物細胞の接着・増殖モニタリングとその応用" Electrochemistry. 67巻・3号. 280-284 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] 海老原史樹文: "生物時計の分子生物学" シュプリンガー・フェアラーク東京, 201 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] H.Shinohara, T.Kusaka, E.Yokota, R.Monden, M.Sisido: "Electron Transfer between Redox Enzymes and Electrodes through the Artificial Redox Proteins and Its Application for Biosensors" Sensors & Actuators Part B (Chemical). (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] H.Shinohara, E.Yokota, K.Yoshitani and M.Sisido: "Design of Artificial Redox Proteins with Specific Binding Activity and Its Application for Biosensor Fabrication" Proceedings of the 4th International Conference on Intelligent Materials (ed.by T.Takagi st al.). 244-245 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Fukuhara, C., N.Suzuki, Y.Matsumoto, Y.Nakayama, K.Aoki, G.Tsujimoto, S.-I.T.Inouye and Y.Masuo: "Day-night variation of pituitary adenylate cyclase-activating polypeptide (PACAP) level in the rat suprachiasmatic nucleus." Neurosci.Lett.229. 49-52 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Shinohara, K., Tominaga, K.and Inouye, S.-I.T.: "Luminance-dependent decrease in vasoactive intestinal polypeptide in the suprachiasmatic nucleus." Neurosci.Lett.251. 21-24 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Ibata Y,Tanaka M,Tamada Y,Hayashi S,Kawakami F,Takamatsu T,Okamura H: "The suprachiasmatic nucleus : Acircadian oscillator." Neuroscientist. 3. 215-225 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Ban Y,Shigeyoshi Y,Okamura H: "Development of circadian VIP rhythm in the rat suprachiasmatic nucleus." J.Neurosci.17. 3920-3931 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Tei H,Okamura H,Shigeyoshi, Y., Fukuhara C,Ozawa R,Hirose M,Sakaki Y: "Circadian oscillation of a mammalian homologue of the Drosophila period gene." Nature. 389. 512-516 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Shigeyoshi Y,Taguchi K,Yamamoto S,Takekida S,Yan L,Tei H,Moriya T,Shibata S,Loros JL,Dunlap JC., Okamura H: "Light-induced resetting of a mammalian circadian clock is associated with rapid induction of the mPer1 transcript." Cell. 91. 1043-1053 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Tamada Y,Tanaka M,Munekawa K,Hayashi S,Okamura H,Kubo T,Hisa Y,Ibata Y: "Neuron-glia interaction in the suprachiasmatic nucleus : a double labelling light and electron microscopic immunocytochemical study in the rat." Brain Res.Bull.45. 281-287 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Takumi T,Matsubara C,Shigeyoshi Y,Taguchi K., Yagita K,Maebayashi Y,Sakakida Y,Okumura K,Takashima N,Okamura H: "A New Mammalian Period Gene Predominantly Expressed in the Suprachiasmatic Nucleus." Genes Cells. 3. 167-176 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Takumi T,Taguchi K,Miyake S,Sakakida Y,Takashima N,Matsubara C,Maebayashi Y,Okumura K,Takekida S,Yamamoto S,Yagita K,Yan L,Young ML,Okamura H: "A light independent oscillatory gene mPer3 in mouse SCN and OVLT." EMBO J. 17. 4753-4759 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Shinohara,K.: "Luminance-dependent decrease invasoactive intestinal polypeptide in the suprachiasmatic nucleus." Neurosci.Lett.251巻. 21-24 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Shinohara,K.: "Phase dependent response of vasoactive intestinal polypeptide to light and darkness in the suprachiasmatic nucleus." Neurosci.Res.(In press).

    • Related Report
      1998 Annual Research Report
  • [Publications] Takumi T.: "A New mammalian period gene predominantly expressed in the suprachiasmatic nucleus." Genes Cells. 3巻. 167-176 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Takumi T.: "A light independent oscillatory gene mPer3 in mouse SCN and OVLT." EMBO J.17巻. 4753-4759 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Takumi,T.: "A mammalian ortholog of Drosophila timeless,highly expressed in SCN and retina,forms a complex with mPER1" Genes Cells. 4巻. 67-75 (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] 篠原寛明: "水晶振動子マイクロバランスを用いる動物細胞の接着・増殖モニタリングとその応用" Electrochemistry. 67巻・3号. 280-284 (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] 海老原史樹文: "生物時計の分子生物学" シュプリンガー・フェアラーク東京, 201 (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Shigeyosi,Y.: "Light-induced resetting of a mammalian circadian clock is associated with rapid induction of the mPer1 transcript" Cell. 91. 1043-1053 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Tei,H.: "Circadian oscillation of a mammalian homolog of the Drosophila period gene" Nature. 389. 512-516 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Ban,Y.: "Development of circadian VIP rhythm in the rat suprachiasmatic nucleus" J.Neurosci.17. 3920-3931 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Shigeyoshi,Y.: "Coexitence of preprosomatostatin and prepro-tachykinin A mRNA in neurons of the rat SCN" Mol.Brain Res.48. 159-163 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Ibata,Y.: "The suprachiasmatic nucleus : A circadian oscillator" Neuroscientist,. 3. 215-225 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kawakami,F.: "Loss of day-night differences in VIP mRNA levels in the suprachiasmatic nucleus of aged rats" Neurosci.Lett.,. 222. 99-102 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Okamura H,Sugano T,Ibata Y: "Reserpine-induced immunocytochemical change of neuropeptide Y in the hypothalamic arcuate nucleus" Neurochem.Res.21. 239-243 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Nakagawa T,UkaiK,OhyamaT Koida M,Okamura H: "Effects of the synthesized growth hormone releasing peptide,KP-102,on growth hormone release in sodium glutamate monohydrate-treated low growth rats" Life Sci.59. 705-712 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Uno T,Hisa Y,Tadaki N,Okamura H,Ibata Y: "Tyrosine hydroxylase immunoreactive cells in the nodose ganglion for the canine larynx" NeuroReport. 7. 1373-1376 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Umehara K,Kataoka,Ogura T Esumi H,Ibata Y,Okamura H: "Comparative distribution of nitric oxide synthase (NOS) in pancreas of the dog and rat" Brain Res.Bull.(In press).

    • Related Report
      1996 Annual Research Report
  • [Publications] Yokoyama C,Okamura H: "Self-injurious behavior and dopaminergic neuron system in neonatal 6-OHDA lesioned rat.1. Dopaminergic neurons and receptors" J.Pharmacol.Exp.Ther.(In press).

    • Related Report
      1996 Annual Research Report
  • [Publications] Okamura H,Murakami T,Yokoyama,Nakamura,Ibata Y: "Self-injurious behavior and dopaminergic neuron system in neonatal 6-OHDA lesioned rat : Intracerebral microinjection of dopamine agonists and antagonists." J.Pharmacol.Exp.Ther.(In press).

    • Related Report
      1996 Annual Research Report
  • [Publications] Hisa Y,Uno T,Tadaki N,Koike,Okamura H,Ibata Y: "Nitric oxide participation in laryngeal innervaiton Vocal Fold Physiology (Davis PJ and Fletcher NF eds)" Singular Publishing Group (Sandiego), 433 (1996)

    • Related Report
      1996 Annual Research Report

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Published: 1997-04-01   Modified: 2016-04-21  

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