@article{f87ca7f0f4064f379d05ab1be2b73fab,
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.",
author = "Sho Wakayama and Shigeki Kiyonaka and Itaru Arai and Wataru Kakegawa and Shinji Matsuda and Keiji Ibata and Nemoto, {Yuri L.} and Akihiro Kusumi and Michisuke Yuzaki and Itaru Hamachi",
note = "Funding Information: We thank Roberto Malinow (University of California, San Diego) for SEP-GluA2 construct, and Eric Gouaux (Oregon Health and Science University) for the S1S2J construct. We also thank K. Akiyoshi and S. Mukai (Kyoto University) for two-photon microscopy, Y. Sato (Carl Zeiss Microscopy Inc.) for confocal microscopy, Y. Mori and S. Sawamura (Kyoto University) for neuronal culture, G. Craven (Imperial College London) for organic synthesis, E. Kusaka (Kyoto University) for NMR measurement, and M. Tsujikawa (Kyoto University) for plasmid construction. This work was funded by a Research Fellowship from the Japan Society for the Promotion of Science (JSPS) for Young Scientists (to S.W. (26-3123)), SUNBOR Grant from Suntory Foundation for Life Sciences (to S.K.), the Takeda Science Foundation (to S.K., W.K. and M.Y.), the Strategic Research Program for Brain Sciences from Japan Agency for Medical Research and Development (AMED) (to W.K.), and the Japan Science and Technology Agency (JST) Core Research for Evolutional Science and Technology (CREST) of Molecular Technologies (to I.H. and M.Y.). Publisher Copyright: {\textcopyright} 2017 The Author(s).",
year = "2017",
month = apr,
day = "7",
doi = "10.1038/ncomms14850",
language = "English",
volume = "8",
journal = "Nature communications",
issn = "2041-1723",
publisher = "Nature Publishing Group",
}