Chemical labelling for visualizing native AMPA receptors in live neurons

Sho Wakayama, Shigeki Kiyonaka (), Itaru Arai, Wataru Kakegawa, Shinji Matsuda, Keiji Ibata, Yuri L. Nemoto, Akihiro Kusumi, Michisuke Yuzaki and Itaru Hamachi ()
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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
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
Michisuke Yuzaki: School of Medicine, Keio University
Itaru Hamachi: Graduate School of Engineering, Kyoto University

Nature Communications, 2017, vol. 8, issue 1, 1-14

Abstract: 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.

Date: 2017
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DOI: 10.1038/ncomms14850

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