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Chemical labelling for visualizing native AMPA receptors in live neurons

Author

Listed:
  • Sho Wakayama

    (Graduate School of Engineering, Kyoto University)

  • Shigeki Kiyonaka

    (Graduate School of Engineering, Kyoto University)

  • Itaru Arai

    (School of Medicine, Keio University)

  • Wataru Kakegawa

    (School of Medicine, Keio University)

  • Shinji Matsuda

    (School of Medicine, Keio University
    Graduate School of Informatics and Engineering, University of Electro-Communication
    PRESTO, Japan Science and Technology Agency)

  • Keiji Ibata

    (School of Medicine, Keio University)

  • Yuri L. Nemoto

    (Institute for Frontier Medical Sciences, Kyoto University)

  • Akihiro Kusumi

    (Institute for Frontier Medical Sciences, Kyoto University
    Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University)

  • Michisuke Yuzaki

    (School of Medicine, Keio University
    CREST(Core Research for Evolutional Science and Technology, JST))

  • Itaru Hamachi

    (Graduate School of Engineering, Kyoto University
    CREST(Core Research for Evolutional Science and Technology, JST))

Abstract

The location and number of neurotransmitter receptors are dynamically regulated at postsynaptic sites. However, currently available methods for visualizing receptor trafficking require the introduction of genetically engineered receptors into neurons, which can disrupt the normal functioning and processing of the original receptor. Here we report a powerful method for visualizing native α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-type glutamate receptors (AMPARs) which are essential for cognitive functions without any genetic manipulation. This is based on a covalent chemical labelling strategy driven by selective ligand-protein recognition to tether small fluorophores to AMPARs using chemical AMPAR modification (CAM) reagents. The high penetrability of CAM reagents enables visualization of native AMPARs deep in brain tissues without affecting receptor function. Moreover, CAM reagents are used to characterize the diffusion dynamics of endogenous AMPARs in both cultured neurons and hippocampal slices. This method will help clarify the involvement of AMPAR trafficking in various neuropsychiatric and neurodevelopmental disorders.

Suggested Citation

  • Sho Wakayama & Shigeki Kiyonaka & Itaru Arai & Wataru Kakegawa & Shinji Matsuda & Keiji Ibata & Yuri L. Nemoto & Akihiro Kusumi & Michisuke Yuzaki & Itaru Hamachi, 2017. "Chemical labelling for visualizing native AMPA receptors in live neurons," Nature Communications, Nature, vol. 8(1), pages 1-14, April.
    • Handle: RePEc:nat:natcom:v:8:y:2017:i:1:d:10.1038_ncomms14850
    DOI: 10.1038/ncomms14850
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