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Measurement of Astrocytic Glutamate Release Using Genetically Encoded Probe Combined with TIRF Microscopy

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Biochemical Approaches for Glutamatergic Neurotransmission

Part of the book series: Neuromethods ((NM,volume 130))

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Abstract

Astrocytes modulate brain function such as memory and cognition by processing neuronal activity through the release of transmitters such as glutamate, ATP and d-serine. Various release mechanisms have been proposed depending on pathophysiological conditions. Among them, inverted transport, ion channels mediated release, and vesicular release. To better understand how cellular interactions alter glutamate release, we used live imaging techniques to monitor the dynamics of glutamate release in various cell culture conditions. For that purpose we combined total internal reflection fluorescence (TIRF) microscopy with a genetically encoded fluorescent glutamate sensor (iGluSnFR) which is an intensity-based glutamate-sensing fluorescent reporter that has a high signal-to-noise ratio. This system allows a direct measurement of glutamate in real time. Our goal is to provide guidelines on how to use this approach and to highlight its pros and cons.

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Author information

Authors and Affiliations

  1. INSERM U1217, CNRS UMR 5310, Institut NeuroMyoGène, Université Claude Bernard Lyon 1, Lyon, France

    Fuyuko Takata-Tsuji, Naura Choulnamountri & Olivier Pascual

  2. Department of Pharmaceutical Care and Health Sciences, Faculty of Pharmaceutical Sciences, Fukuoka University, Fukuoka, Japan

    Fuyuko Takata-Tsuji

  3. ENS de Lyon, Université Claude Bernard Lyon 1, CNRS, Laboratoire de Physique, 69342, Lyon, France

    Christophe Place

Authors
  1. Fuyuko Takata-Tsuji
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  2. Naura Choulnamountri
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  3. Christophe Place
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  4. Olivier Pascual
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Corresponding author

Correspondence to Olivier Pascual .

Editor information

Editors and Affiliations

  1. Lyon Neuroscience Research Center, University Lyon 1, Lyon, France

    Sandrine Parrot

  2. Lyon Neuroscience Research Center, University Lyon 1, Lyon, France

    Luc Denoroy

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Takata-Tsuji, F., Choulnamountri, N., Place, C., Pascual, O. (2018). Measurement of Astrocytic Glutamate Release Using Genetically Encoded Probe Combined with TIRF Microscopy. In: Parrot, S., Denoroy, L. (eds) Biochemical Approaches for Glutamatergic Neurotransmission. Neuromethods, vol 130. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7228-9_6

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  • DOI: https://doi.org/10.1007/978-1-4939-7228-9_6

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7227-2

  • Online ISBN: 978-1-4939-7228-9

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