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, Itaru Hamachi

Research output: Contribution to journalArticle

25 Citations (Scopus)

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.

Original languageEnglish
Article number14850
JournalNature Communications
Volume8
DOIs
Publication statusPublished - 2017 Apr 7

Fingerprint

alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid
AMPA Receptors
Glutamate Receptors
neurons
Labeling
marking
Neurons
glutamates
Chemical modification
reagents
Neurotransmitter Receptor
Fluorophores
Cognition
Brain
Visualization
neurotransmitters
Tissue
Ligands
Acids
Processing

ASJC Scopus subject areas

  • Chemistry(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Physics and Astronomy(all)

Cite this

Chemical labelling for visualizing native AMPA receptors in live neurons. / Wakayama, Sho; Kiyonaka, Shigeki; Arai, Itaru; Kakegawa, Wataru; Matsuda, Shinji; Ibata, Keiji; Nemoto, Yuri L.; Kusumi, Akihiro; Yuzaki, Michisuke; Hamachi, Itaru.

In: Nature Communications, Vol. 8, 14850, 07.04.2017.

Research output: Contribution to journalArticle

Wakayama, S, Kiyonaka, S, Arai, I, Kakegawa, W, Matsuda, S, Ibata, K, Nemoto, YL, Kusumi, A, Yuzaki, M & Hamachi, I 2017, 'Chemical labelling for visualizing native AMPA receptors in live neurons', Nature Communications, vol. 8, 14850. https://doi.org/10.1038/ncomms14850
Wakayama, Sho ; Kiyonaka, Shigeki ; Arai, Itaru ; Kakegawa, Wataru ; Matsuda, Shinji ; Ibata, Keiji ; Nemoto, Yuri L. ; Kusumi, Akihiro ; Yuzaki, Michisuke ; Hamachi, Itaru. / Chemical labelling for visualizing native AMPA receptors in live neurons. In: Nature Communications. 2017 ; Vol. 8.
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