title: Chemical labelling for visualizing native AMPA receptors in live neurons.
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.
authors.affiliation: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Nishikyo-ku 615-8510, Japan.
authors.forename: Sho
authors.initials: S
authors.lastname: Wakayama
authors.affiliation: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Nishikyo-ku 615-8510, Japan.
authors.forename: Shigeki
authors.initials: S
authors.lastname: Kiyonaka
authors.affiliation: Department of Physiology, School of Medicine, Keio University, Tokyo 160-8582, Japan.
authors.forename: Itaru
authors.initials: I
authors.lastname: Arai
authors.affiliation: Department of Physiology, School of Medicine, Keio University, Tokyo 160-8582, Japan.
authors.forename: Wataru
authors.initials: W
authors.lastname: Kakegawa
authors.affiliation: Department of Physiology, School of Medicine, Keio University, Tokyo 160-8582, Japan.|Department of Engineering Science, Graduate School of Informatics and Engineering, University of Electro-Communication, Tokyo 182-8585, Japan.|PRESTO, Japan Science and Technology Agency, Saitama 332-0012, Japan.
authors.forename: Shinji
authors.initials: S
authors.lastname: Matsuda
authors.affiliation: Department of Physiology, School of Medicine, Keio University, Tokyo 160-8582, Japan.
authors.forename: Keiji
authors.initials: K
authors.lastname: Ibata
authors.affiliation: Institute for Frontier Medical Sciences, Kyoto University, Kyoto 606-8507, Japan.
authors.forename: Yuri L
authors.initials: YL
authors.lastname: Nemoto
authors.affiliation: Institute for Frontier Medical Sciences, Kyoto University, Kyoto 606-8507, Japan.|Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, Kyoto 606-8507, Japan.
authors.forename: Akihiro
authors.initials: A
authors.lastname: Kusumi
authors.affiliation: Department of Physiology, School of Medicine, Keio University, Tokyo 160-8582, Japan.|CREST(Core Research for Evolutional Science and Technology, JST), Chiyodaku, Tokyo 102-0075, Japan.
authors.forename: Michisuke
authors.initials: M
authors.lastname: Yuzaki
authors.affiliation: Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Nishikyo-ku 615-8510, Japan.|CREST(Core Research for Evolutional Science and Technology, JST), Chiyodaku, Tokyo 102-0075, Japan.
authors.forename: Itaru
authors.initials: I
authors.lastname: Hamachi
mesh_terms: D000818:Animals; D057809:HEK293 Cells; D006624:Hippocampus / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D007202:Indicators and Reagents / Q000737:chemistry; D051379:Mice; D008813:Mice, Inbred ICR; D009474:Neurons / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D018091:Receptors, AMPA / Q000378:metabolism*
chemical_list: D007202:Indicators and Reagents; D018091:Receptors, AMPA