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An Enzyme-entrapped Agarose Gel for Visualization of Ischemia-induced L-Glutamate Fluxes in Hippocampal Slices in a Flow System

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Abstract

An agarose gel slip containing L-glutamate oxidase (GluOx), horseradish peroxidase (HRP) and a dye DA-64 is proposed as a tool for visualizing ischemia-induced L-glutamate release in hippocampal slices in a flow system. The agarose slip with a detection limit of 6.0 ± 0.8 μmol L−1 for L-glutamate enabled us to visualize L-glutamate fluxes in a flow system. The leak of a dye from the agarose gel was negligible and a diffusion blur due to spreading of Bindshedler’s Green (BG) within the gel was suppressed. Monitoring the time-dependent change of ischemia-induced L-glutamate fluxes at neuronal regions CA1, DG and CA3 of hippocampal slices is demonstrated.

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Acknowledgments

The authors thank Mr. N. Tamura for his experimental help. This work was supported in part by Grants-in-Aid for Scientific Research (No. 21350045) from the Ministry of Education, Culture, Sports, Science and Technology, Japan.

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Authors and Affiliations

  1. Department of Chemistry, College of Humanities and Sciences, Nihon University, Sakurajousui, Setagaya, Tokyo, 156-8550, Japan

    Kazuhisa Tanaka & Masao Sugawara

  2. Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, Tokyo, 192-0329, Japan

    Atushi Shoji

Authors
  1. Kazuhisa Tanaka
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  2. Atushi Shoji
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  3. Masao Sugawara
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Correspondence to Masao Sugawara.

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Tanaka, K., Shoji, A. & Sugawara, M. An Enzyme-entrapped Agarose Gel for Visualization of Ischemia-induced L-Glutamate Fluxes in Hippocampal Slices in a Flow System. ANAL. SCI. 31, 321–325 (2015). https://doi.org/10.2116/analsci.31.321

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  • DOI: https://doi.org/10.2116/analsci.31.321

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