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Development of in situ detection methods for substances in brain by using bioelements and bilayer lipid membranes

Research Project

Project/Area Number 15350050
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Analytical chemistry
Research InstitutionNihon University

Principal Investigator

SUGAWARA Masao  Nihon University, College of Humanities and Sciences, Professor, 文理学部, 教授 (50002176)

Project Period (FY) 2003 – 2005
Project Status Completed (Fiscal Year 2005)
Budget Amount *help
¥13,300,000 (Direct Cost: ¥13,300,000)
Fiscal Year 2005: ¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 2004: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 2003: ¥9,100,000 (Direct Cost: ¥9,100,000)
KeywordsIn situ detection / neurotransmitter / electrochemical sensor / chemical imaging / arachidonic acid / glutamate / bilayer lipid membrane sensor / patch sensor / バイオ素子 / サブスタンスP / カルモジュリン / リポソームイムノアッセイ / キャピラリー酵素センサー / 表面プラズモン共鳴法 / DNA損傷 / リポソーム / 逆行性神経伝達物質 / マウス海馬 / グルタミン酸センサー / ガラスキャピラリーセンサー / マウス脳スライス / 脂質二分子膜 / 酵素膜 / 可視化イメージング / 虚血
Research Abstract

For the design of in situ sensing systems for biologically important substances that participates in neuronal communication in acute mouse brain slices, the use of artificial and natural bilayer lipid membranes and biological elements has be investigated. A glass capillary-based enzyme electrode was designed to detect L-glutamate that is released under glucose-free and oxygen-deficient conditions (Ischemia) by using glutamate oxidase and a gall capillary having the ability of sampling a small volume of extracelluar solution. The sensor applied to acute mouse brain slices showed that release of L-glutamate is neuronal region-dependent and the amount of L-glutamate release increases in the order of CA1【approximately equal】CA3>DG. This order was in agreement with those obtained by differential imaging of L-glutamate fluxes using enzyme-immobilized membrane developed newly in this research project. A patch sensor containing NMDA receptors was developed for detecting GABA-induced release of glutamate in acute hippocampal slices. The results showed that in the CA1 region, glutamate release by GABA stimulation is induced via GABA_A receptor and in the CA3 region, release of glutamate was not observed due to inhibitory effect of GABA_B receptor. An excised patch membrane sensor for arachidonic acid was also developed and applied to detect L-glutamate-stimulated release of arachidonic acid. The results exhibited that release of arachidonic acid increases in the order of CA3>CA1【approximately equal】DG. Also, bilayer lipid membranes (BLMs) containing a single gramicidin channel was prepared by the tip-dip method. The BLMs exhibited current responses, corresponding to modulation of gramicidin monomer/dimer kinetics caused by the binding of analytes to membrane bound receptor sites in the BLMs. The BLMs were used for detecting antibodies.

Report

(4 results)
  • 2005 Annual Research Report   Final Research Report Summary
  • 2004 Annual Research Report
  • 2003 Annual Research Report
  • Research Products

    (35 results)

All 2006 2005 2004 2003 Other

All Journal Article (28 results) Book (3 results) Publications (4 results)

  • [Journal Article] Detection of GABA-induced Glutamate Release in Acute Mouse Hippocampal Slices with a Patch Sensor2006

    • Author(s)
      M.Shimane
    • Journal Title

      Anal. Biochem. in press

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Detection of GABA-induced Glutamate release in acute mouse hippocampal slices with a patch sensor2006

    • Author(s)
      M.Shimane, K.Miyagawa, M.Sugawara
    • Journal Title

      Anal.Biochem. (in press)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Nitrogenous Synergists Induced Potentiometric Response to Metal Ions with Polymeric Liquid Membranes Containing Thenoyltrifluoroacetone as an Ionophore2006

    • Author(s)
      Y.Itoh, Y.Ueda, M.Sugawara, M.Kataoka, H.Sato, Y.Umezawa
    • Journal Title

      Anal. Sci. 22

      Pages: 219-223

    • Related Report
      2005 Annual Research Report
  • [Journal Article] A surface Plasmon Resonance Sensor for Substance P Using Gold-modified Calmodulin and Melittin2005

    • Author(s)
      T.Karasawa
    • Journal Title

      Anal. Sci 21

      Pages: 1431-1431

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Real Time Monitoring of L-Glutamate Release from Mouse Brain Slices under Ischemia with a Glass Capillary-Based Enzyme Electrode2005

    • Author(s)
      N.Nakamura
    • Journal Title

      Anal. Bioanal. Chem. 330

      Pages: 660-660

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Real Time Monitoring of L-Glutamate Release from Mouse Brain Slices under Ischemia with a Glass Capillary-Based Enzyme Electrode2005

    • Author(s)
      N.Nakamura, K.Negishi, A.Hirano, M.Sugawara
    • Journal Title

      Anal.Bioanal.Chem. 330

      Pages: 660-667

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Design and Application of Planar Bilayer Lipid Membranes Containing Biological Ion Channels for Chemical Sensing2005

    • Author(s)
      M.Sugawara, A.Hirano
    • Journal Title

      Advances in Planar Lipid Bilayers and Liposomes, Chapter 8 (Eds.H.T.Tien and A.Ottova)(Elsevier) Vol.1

      Pages: 221-245

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Real Time Monitoring of L-Glutamate Release from Mouse Brain Slices under Ischemia with a Glass Capillary-Based Enzyme Electrode2005

    • Author(s)
      N.Nakamura, K.Negishi, A.Hirano, M.Sugawara
    • Journal Title

      Anal. Bioanal. Chem. 330

      Pages: 660-667

    • Related Report
      2005 Annual Research Report
  • [Journal Article] A surface Plasmon Resonance Sensor for Substance P Using Gold-modified Calmodulin and Melittin2005

    • Author(s)
      T.Karasawa, M.Sugawara
    • Journal Title

      Anal. Sci. 21

      Pages: 1431-1436

    • Related Report
      2005 Annual Research Report
  • [Journal Article] An Excised Patch Membrane Sensor for Arachidonic Acid Released in Mouse Hippocampal Slices under Stimulation of L-glutamate2004

    • Author(s)
      H.Saitoh
    • Journal Title

      Anal. Biochem. 329

      Pages: 163-163

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Single-Channel Recordings of Gramicidin at Agarose-Supported Bilayer Lipid Membranes Formed by the Tip-Dip and Painting Methods2004

    • Author(s)
      Y.Matsuno
    • Journal Title

      Anal. Sci. 20 1217

      Pages: 1217-1217

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] 毛管作用に基づく超微小ガラスキャピラリーセンサーの開発と哺乳類大脳内のグルタミン酸濃度測定への応用2004

    • Author(s)
      平野愛弓
    • Journal Title

      ぶんせき

      Pages: 652-652

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] An Excised Patch Membrane Sensor for Arachidonic Acid Released in Mouse Hippocampal Slices under Stimulation of L-glutamate.2004

    • Author(s)
      H.Saitoh, Y.Namatame, A.Hirano, M.Sugawara
    • Journal Title

      Anal.Biochem. 329

      Pages: 163-172

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Single-Channel Recordings of Gramicidin at Agarose-Supported Bilayer Lipid Membranes Formed by the Tip-Dip and Painting Methods2004

    • Author(s)
      Y.Matsuno, C.Osono, A.Hirano, M.Sugawara
    • Journal Title

      Anal.Sci. 20

      Pages: 1217-1221

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] (2005): A surface Plasmon Resonance Sensor for Substance P Using Gold-modified Calmodulin and Melittin2004

    • Author(s)
      T.Karasawa, M.Sugawara
    • Journal Title

      Anal.Sci. 21

      Pages: 1431-1436

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Development of an ultramicro glass capillary sensor and its application to detection of L-glutamate in brain (in Japanese)2004

    • Author(s)
      A.Hirano, M.Sugawara
    • Journal Title

      BUNSEKI

      Pages: 652-654

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] An Excised Patch Membrane Sensor for Arachidonic Acid Released in Mouse Hippocampal Slices under Stimulation of L-glutamate2004

    • Author(s)
      H.Saitoh, Y.Namatame, A.Hirano, M.Sugawara
    • Journal Title

      Anal.Biochem. 329

      Pages: 163-172

    • Related Report
      2004 Annual Research Report
  • [Journal Article] A Dot-Blot Method for Fluorometric Detection of Apurinic Sites in DNA Using an Avidin-Coated Glass Plate2004

    • Author(s)
      H.Yanagisawa, A.Hirano, M.Sugawara
    • Journal Title

      Anal.Biochem. 332

      Pages: 358-367

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Single-Channel Recordings of Gramicidin at Agarose-Suppted or Bilayer Lipid Membranes Formed by the Tip-Dip and Painting Methods2004

    • Author(s)
      Y.Matsuno, C.Osono, A.Hirano, M.Sugawara
    • Journal Title

      Anal.Sci. 20

      Pages: 1217-1221

    • Related Report
      2004 Annual Research Report
  • [Journal Article] 毛管作用に基づく超微小ガラスキャピラリーセンサーの開発と哺乳類大脳内のグルタミン酸濃度測定への応用2004

    • Author(s)
      平野愛弓, 菅原正雄
    • Journal Title

      ぶんせき

      Pages: 652-654

    • NAID

      10013771313

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Imaging of L-Glutamate Fluxes in Mouse Brain Slices Based on an Enzyme-Based Membrane Combined with a Differential-Image Analysis2003

    • Author(s)
      A.Hirano
    • Journal Title

      Anal. Chem. 75

      Pages: 3775-3775

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] A Single-Channel Sensor Based on Gramicidin Controlled by Molecular Recognition at Bilayer Lipid Membranes Containing Receptor2003

    • Author(s)
      A.Hirano
    • Journal Title

      Biosensors & Bioelectronics 18

      Pages: 973-973

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Arachidonic Acid-Induced Channel and Carrier-Type Ion transport across Planar Bilayer Lipid Membranes2003

    • Author(s)
      A.Hirano
    • Journal Title

      Anal. Sci. 19

      Pages: 191-191

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] A Glass Capillary Microelectrode Based on Capillarity and Its Application to Detection of L-glutamate Release from Mouse Brain Slices2003

    • Author(s)
      K.Nakajima
    • Journal Title

      Anal. Sci. 19

      Pages: 55-55

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Imaging of L-Glutamate Fluxes in Mouse Brain Slices Based on an Enzyme-Based Membrane Combined with a Differential-Image Analysis2003

    • Author(s)
      A.Hirano, N.Moridera, M.Akashi M.Saito, M.Sugawara
    • Journal Title

      Anal.Chem. 75

      Pages: 3775-3783

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] A Single-Channel Sensor Based on Gramicidin Controlled by Molecular Recognition at Bilayer Lipid Membranes Containing Receptor2003

    • Author(s)
      A.Hirano, M.Wakabayashi, Y.Matsuno, M.Sugawara
    • Journal Title

      sBiosensors & Bioelectronics 18

      Pages: 973-983

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Arachidonic Acid-Induced Channel and Carrier-Type Ion transport across Planar Bilayer Lipid Membranes2003

    • Author(s)
      A.Hirano, Y.Namatame, E.Wakaizumi, Y.Matsuno, M.Sugawara
    • Journal Title

      Anal.Sci. 19

      Pages: 191-197

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] A Glass Capillary Microelectrode Based on Capillarity and Its Application to Detection of L-glutamate Release from Mouse Brain Slices2003

    • Author(s)
      K.Nakajima, T.Yamagiwa, A.Hirano, M.Sugawara
    • Journal Title

      Anal.Sci. 19

      Pages: 55-60

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Book] Advances in Planar Lipid Bilayers and Liposomes (分担執筆)2005

    • Author(s)
      M.Sugawara
    • Total Pages
      24
    • Publisher
      Elsevier
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Book] Planar Lipid Bilayers and Liposomes(分担執筆)、 Vol.1 ed. by Tien and Ottova2005

    • Author(s)
      M.Sugawara, A.Hirano
    • Total Pages
      24
    • Publisher
      Elsevier
    • Related Report
      2005 Annual Research Report
  • [Book] Advances in Planar Lipid Bilayers and Liposomes-Recent Progress-2003/4,Eds.H.T.Tien and A.Ottova2005

    • Author(s)
      M.Sugawara, A.Hirano
    • Publisher
      Elsevier(In press)
    • Related Report
      2004 Annual Research Report
  • [Publications] K.Nakajima: "A Glass Capillary Microelectrode Based on Capillarity and Its Application to Detection of L-glutamate Release from Mouse Brain Slices"Anal.Sci.. 19. 55-60 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] A.Hirano: "Arachidonic Acid-Induced Channel and Carrier-Type Ion transport across Planar Bilayer Lipid Membranes"Anal.Sci.. 19. 191-197 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] A.Hirano: "A Single-Channel Sensor Based on Gramicidin Controlled by Molecular Recognition at Bilayer Lipid Membranes Containing Receptor"Biosensors & Bioelectronics. 18. 973-983 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] A.Hirano: "Imaging of L-Glutamate Fluxes in Mouse Brain Slices Based on an Enzyme-Based Membrane Combined with a Differential-Image Analysis"Anal.Chem.. 75. 3775-3783 (2003)

    • Related Report
      2003 Annual Research Report

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Published: 2003-04-01   Modified: 2016-04-21  

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