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ESTABLISHMENT OF AN ONLINE ASSAY SYSTEM OF INTRACEREBRALLY RELEASED GLUTAMATE BY MEANS OF MICROBIOCENSOR

Research Project

Project/Area Number 06557038
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section試験
Research Field 内科学一般
Research InstitutionUniversity of the Air

Principal Investigator

KITO Shozo  University of the Air, Liberal Arts, Professor, 教養学部, 教授 (00010140)

Co-Investigator(Kenkyū-buntansha) WATANABE Yasuhiro  National Defense Medical College, Professor, 教授 (90127324)
SEMBA Jun'ichi  Univ.of the Air, Liberal Arts, Associate Professor, 教養学部, 助教授 (30183429)
Project Period (FY) 1994 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥16,800,000 (Direct Cost: ¥16,800,000)
Fiscal Year 1996: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1995: ¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 1994: ¥10,200,000 (Direct Cost: ¥10,200,000)
Keywordsglutamate / platinum black sensor / hydrogen peroxide / sensor output / ascorbic acid / platinum tube / glutamate oxidase / glutamine / glutamine oxidase / ptチューブ / カイニン酸 / glucocorticoid / nicotine / dexamethasone / in situ hybridization / in vivo microdialysis / estrogen / Ca^<2+>
Research Abstract

This study was undertaken to assay concentrations of extracellular L-glutamate release from imtracerebral neurons in vivo with a small-volume online sensor made of platinum black consisted of microdialysis probe fixed at the manipulator.
As the first step, in vitro studies to get a linear calibration curve of L-glutamate determined with sensor output with use of the BAS100B/W workstation and low current module to increase the sensitivity and lower the noise level.
The generated hydrogen peroxide is oxidized electrochemically at a Pt electrode held at 600mV and the oxidized current was monitored as a linear relationship between sensor output and glutamate concentration.
As for in vivo experiments, we tried to remove extracellular ascorbic acid which interfere the assay system. We also performed experiments using carbon bulk censer with Os poly meditator instead of platinum black. In this method, the potential of the electrode can be held at 0 mV so that there is no interference by either catecholamine or ascorbic acid.
An another problem in vivo online measurement is that we can not avoid, contamination with glutamine from this assay system, we need a glutamate oxidase which is extremely specific to glutamate. Purification of this kind of enzyme is currently in progress. With this enzyme coming up, establishment of the whole assay system is completed.

Report

(4 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report
  • 1994 Annual Research Report
  • Research Products

    (70 results)

All Other

All Publications (70 results)

  • [Publications] 新郷 明子: "ゲルシフトアッセイからみたエストロゲンとニコチンのクロストーク" 医学と生物学. 134/1. 3-7 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Semba,J.: "Nicotine protects against the dexamrthasone potentiation of kainic acid-Induced neurotoxicity in cultured hippocampal neurons." Brain Research. 735. 335-338 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Semba,J.: "Differential expression of c-fos mRNA in rat prefrontal cortex,striatum,N.accumbens and lateral septum after typical and atypical antipsychotics:an in situ hybridization study." Neurochemistry International. 29/4. 435-442 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 新郷明子: "カイニン酸傷害脳に対するestrogenの影響" 神経化学. 35/6. 510-511 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 新郷明子: "エストロゲンと脳可塑性に関する研究" 医学と生物学. 133/5. 189-193 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 新郷明子: "カイニン酸とAP-1 DNA結合活性-その時間的推移-" 医学と生物学. 133/5. 195-199 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Semba,J.: "Characterization of extracellular amino acids in striatum of freely moving rats by in vivo microdialysis." Journal of Neural Transmission. 100. 39-52 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Semba,J.: "Different expression of c-fos mRNA in rat striatum,N.accumbens and prefrontal cortex after haioperidol,sulpiride and clozapine:an in situ hybridization study." Japanese Journal of Pharmacology. 67. 143 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Kito,S.: "Effects of estrogen on culturedlimbic neurons." Japanese Journal of Pharmacology. 67. 157 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Semba,J.: "N^G-monomethyl-L-arginine,ainhibitor of nitric oxide synthase,increases extracellular GABA in the striatum of the freely moving rat." Neuro Report. 6. 1426-1428 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Semba,J.: "Behavioural and neurochemical effect of OPC-14597,a novel antipsychotic drug,on dopaminergic mechanisms in rat brain." Neuropharmacology. 34. 785-791 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Semba,J.: "Effect of systemic dexamethasone on extracellular amino acids(glutamate,aspartate,glycine and GABA)and acetylcholine release in rat hippocampus in vivo microdialysis." Functional Neurology. 10. 17-21 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Semba,J.: "Characterization of extracellular amino acids in striutum of freely moving rat using in vivo microdialysis." Japanese Journal of Pharmacology. 64. 229 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Semba,J.: "Antiplleptic drug pharmacokinetics and neuropharmacolokinetics in individual rats by repetitive withdrawal of blood and cerebrospinal fruid milacemide." Br.J.Pharmacol.108. 1117-1124 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Nishio,E.: "Glucose-induced down-regulation of NO production and inducible NOS expression in cultured rat aortic vascular smooth muscle cells:role of protein kinase C." Biochem Biophys Res Commun.229. 857-63 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Tsuneoka,Y.: "Genotype analysis of the CYP2C19 gene in the Japanese population." Life Sci.59. 1711-5 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Shingo, A.S: "Estrogen-nicotine cross-talk in rat brain" Medicine and Biology. 134. 3-7 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Semba, J.: "Nicotine protects against the dexamrthasone potentiation of kainic acid-induced neurotoxicity in cultured hippocampal neurons." Brain Research. 735. 336-338 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Semba, J.: "Differential expression of c-fos mRNA in rat prefrontal cortex, striatum, N.Accumbens and lateral septum after typical and atypical antipsychotics : an in situ hydridization study." Neurochemistry International. 29. 435-442 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Shingo, A.S.: "Effect of estrogen on the kainic acid-injured brain" J.J.of Neurochemistry. 35. 510-511 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Shingo, A.S.: "Estrogen plays a role in neuronal plasticity at injured brains" Medicine and Biology. 133/5. 189-193 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Shingo, A.S.: "Studies on kainic acid induced AP-1 DNA binding activities - the time course-" Medicine and Biology. 133. 195-199 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Semba, J.: "Characterization of extracellular amino acids in striatum of freely moving rats by in vivo microdialysis." Journal of Neural Transmission. 100. 39-52 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Differential expression of c-fos mRNA in rat striatum, N.accumbens and prefrontal cortex after haloperidol, sulpiride and clozapine : an in situ hybridization study.: "Japanese Journal of Pharmacology" 67. 143 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Kito, S.: "Effects of estrogen on cultured limbic neurons." Japanese Journal of Pharmacology. 67. 157 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Semba, J.: "N^G-monomethyl-L-arginine, ainhibitor of nitric oxide synthase, increases extracellular GABA in the striatum of the freely moving rat." Neuro Report. 6. 1426-8 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Semba, J.: "Behavioral and neurochemical effect of OPC-14597, a novel antipsychotic drug, on dopaminergic mechanisms in rat brain." Neuropharmacology. 34. 785-91 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Semba, J.: "Effect of systemic dexamethasone on extracellular amino acids (glutamate, aspartate, glycine and GABA) and acetylcholine release in rat hippocampus in vivo microdialysis." Functional Neurology. 10. 17-21 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Semba, J.: "Characterization of extracellular amino acids in striatum o freely moving rat using in vivo microdialysis." Japanese Journal of pharmacology. 64. 229 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Semba, J.: "Antipileptic drug pharmacokinetics and neuropharmacolokinetics in individual rats by repetitive withdrawal of blood and cerebrospinal fruid milacemide." Br.J.Pharmacol.108. 1117-1124 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Nishio, E.: "Glucose-induced down-regulation of NO production and inducible NOS expression in cultured rat aortic vascular smooth muscle cells : role of protein kinase C." Biochem Biophys Res Commun.229. 857-63 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Tsuneoka, Y.: "Genotype analysis of the CYP2C19 gene in the Japanese population." Biochem Biophys Res Commun.59. 1711-5 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 新郷明子: "ゲルシフトアッセイからみたエストロゲンとニコチンのクロストーク" 医学と生物学. 134/1. 3-7 (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Semba,J.: "Nicotine protects against the dexamrthasone potentiation of kainic acld-induced neurotoxicity in cultured hippocampal neurons." Brain Research. 735. 335-338 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Semba,J.: "Differential expression of c-fos mRNA in rat prefrontal cortex,striatum,N.accumbens and lateral septum after typical and atypical antipsychotics : an in situ hybridization study." Neurochemistry International. 29/4. 435-442 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 新郷明子: "カイニン酸傷害脳に対するestrogenの影響" 神経化学. 35/6. 510-511 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 新郷明子: "エストロゲンと脳可塑性に関する研究" 医学と生物学. 133/5. 189-193 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 新郷明子: "カイニン酸とAP-1 DNA結合活性-その時間的推移-" 医学と生物学. 133/5. 195-199 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Semba,J.: "Characterization of extracellular amino acids in striatum of freely moving rats by in vivo microdialysis." Journal of Neural Transmission. 100. 39-52 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] Semba,J.: "Different expression of c-fos mRNA in rat striatum,N.accumbens and prefrontal cortex after haloperidol,sulpiride and clozapine : an in situ hybridization study." Japanese Journal of Pharmacology. 67. 143 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] Kito,S.: "Effects of estrogen on cultured limbic neurons." Japanese Journal of Pharmacology. 67. 157 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] Semba,J.: "N^G-monomethyl-_L-arginine,alnhibitor of nitric oxide synthase,increases extracellular GABA in the striatum of the freely moving rat." Neuro Repo t. 6. 1426-1428 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] Semba,J.: "Behavioural and neurochemlcai effect of OPC-14597,a novel antipsychotic drug,on dopaminergic mechanisms in rat brain." Neuropharmacology. 34. 785-791 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] Semba,J.: "Effect of systemic dexamethasone on extracellular amino acids(glutamate,aspartate,glycine and GABA) and acetylcholine release in rat hippocampus in vivo microdialysis." Functional Neurology. 10. 17-21 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] Semba, J.: "Differential expression of c-fos mRNA in rat prefrontal cortex, striatum, N. accoumbens and lateral septum after typical and atypical antipsychotics: an in situ hybridization study." Neurochemistry Interational. (in press).

    • Related Report
      1995 Annual Research Report
  • [Publications] Semba, J.: "Differential expression of c-fos mRNA in rat striatum, N. accumbens and prefrontal cortex after haloperidol, sulpiride and clozapine: an in situ hybridization study." Japanese Journal of Pharmacology. 67. 143- (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Kito, S.: "Effects of estrogen on cultured limbic neurons." Japanese Journal of Pharmacology. 67. 157- (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Semba, J.: "N^g-monomethyl-_L-arginine, an inhibitor of nitric oxide synthase, increases extracellular GABA in the striatum of the freely moving rat." Neuro Report. 34. 1426-1428 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Semba, J.: "Behavioural and neurochemical effects of OPC-14597, a novel antipsychotic drug, on dopaminergic mechanisms in rat brain." Neuropharmacology. 34. 785-791 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Semba, J.: "Effects of systemic dexamethasone on extracellular amino acids (glutamate, aspartate, glycine and GABA) and acetylcholine release in rat hippocampus in vivo microdialysis." Functional Neurology. 10. 17-21 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Semba, J.: "Characterization of extracellular amino acids in striatum of freely moving rat using in vivo microdialysis." Japanese Journal of Pharmacology. 64. 229- (1994)

    • Related Report
      1995 Annual Research Report
  • [Publications] Kito, S.: "Effects of nicotine on glucocorticoid toxicity on hippocampal nerouns." Neurochemical Research. 19. 888-889 (1994)

    • Related Report
      1995 Annual Research Report
  • [Publications] Kito, S.: "Effects of glucocorticoid on Hippocampal Neurons: in vivo and in vitro studies" Neurochemical Research. 18. 832- (1993)

    • Related Report
      1995 Annual Research Report
  • [Publications] Semba, J.: "Antiepileptic drug pharmacokinetics and neuropharmacokinetics in individual rats by repetitive withdrawal of blood and cerebrospinal fluid milacemide." Br. J. Pharmacol.108. 1117-1124 (1993)

    • Related Report
      1995 Annual Research Report
  • [Publications] Sakata, H.: "Antiarrhytimic durugs, clofilium and cibenzoline are potent inhibitors of glibenclamide-sensitive K_+ currents in Xenopus oocytes." Br. J. Pharmacol.109. 866-872 (1993)

    • Related Report
      1995 Annual Research Report
  • [Publications] Semba,J.: "Behavioral and neurochemical effects of OPC-14597,a novel antipsychotics drugs,on dopaminergic mechanisms in ratbrain." Neuropharmacology. (in press).

    • Related Report
      1994 Annual Research Report
  • [Publications] Semba,J.: "Effect of systemic dexamethasone on extracellular ammino acids(glutamate,aspartate,glycine and GABA)and acetylcholine release in rat hippocampus in vivo microdialysis." Functional Neurology. (in press).

    • Related Report
      1994 Annual Research Report
  • [Publications] Semba,J.: "Characterisation of extracellular amino acids in striatum of freely moving rats by in vivo microdialysis." J.of Neural Transmission. (in press).

    • Related Report
      1994 Annual Research Report
  • [Publications] Kito,S.: "Effect of nicotine on glucocorticoid toxicity on hippocampal neurons." Neurochemical Research. 19. 888-889 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Kito,S.: "Effects of glucocorticoid on hippocampal neurons:in vivo and in vitro studies." Neurochemical Research. 18. 832

    • Related Report
      1994 Annual Research Report
  • [Publications] Miyoshi,R.: "Effect of caerulein on expression of the immediate early genes,c-fos and zif/268 in the rat brain." Neurocience Letters. 145. 205-208 (1992)

    • Related Report
      1994 Annual Research Report
  • [Publications] Semba,J.: "Behavioral and neurochemical effects of OPC-14597,a novel antipsychotics drugs,on dopaminergic mechanisms in rat brain." Neuropharmacology. (in press).

    • Related Report
      1994 Annual Research Report
  • [Publications] Semba,J.: "Effect of systemic dexamethasone on extracellular amino acids(glutamate,aspartate,glycine and GABA)and acetylcholine release in re thippocampus in vivo microdialysis." Functional Naurology. (in press).

    • Related Report
      1994 Annual Research Report
  • [Publications] Semba,J.: "Characterisation of extracellular amino acids in striatum of freely moving rats by in vivo microdialysis." J.of Neural Transmission. (in press9.

    • Related Report
      1994 Annual Research Report
  • [Publications] Kito,S.: "Effect of nicotine on glucocorticoid toxicity on hippocampal neurons." Neurochemical Research. 19. 888-889 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Semba,J.: "Antiepileptic drug pharmacokinetics and neuropharmacokinetics in individual rats by repetitive withdrawal of blood and cerebrospinal fluid milacemide." Br.J.Pharmacol.108. 1117-1124 (1993)

    • Related Report
      1994 Annual Research Report
  • [Publications] Semba,J.: "Milacemide effects on the temporal interrelationship of amino acids and monoamine metabolites in rat cervbrospinal fluvd." European Journal of Pharmacology. 230. 321-326 (1993)

    • Related Report
      1994 Annual Research Report
  • [Publications] Sakuta.H.: "ntiarrhythmic drugs,clofilium and cibenzotine are potent inhibitors of glibenclamide-sensitive K+currents in Xenopus oocytes." Br.J.Pharmacol.109. 866-872 (1993)

    • Related Report
      1994 Annual Research Report
  • [Publications] Wang,X.B.: "In situ DNA^protein binding:a novel method for detecting DNA-binding activity of transcription factor in brain." Neuroscience Letter. 146. 25-28 (1992)

    • Related Report
      1994 Annual Research Report
  • [Publications] Sakagoshi,N.: "Alterations in the bata-adrenergic receptor system after hypothemic ischemic in hearts with preischemic beta-receptor desensitization." J.Surgical Research. 53. 66-73 (1993)

    • Related Report
      1994 Annual Research Report

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Published: 1994-04-01   Modified: 2016-04-21  

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