Journal Article 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

82017-04-07 , Springer Nature
記憶や学習に関わる神経伝達物質受容体の新規蛍光標識法を開発--神経疾患や精神疾患の診断への活用に期待--. 京都大学プレスリリース. 2017-04-10.
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.

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