RT Journal Article
SR Electronic
T1 Ligand-directed two-step labeling to quantify neuronal glutamate receptor trafficking
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.05.20.105296
DO 10.1101/2020.05.20.105296
A1 Kento Ojima
A1 Kazuki Shiraiwa
A1 Tomohiro Doura
A1 Mikiko Takato
A1 Kazuhiro Komatsu
A1 Michisuke Yuzaki
A1 Itaru Hamachi
A1 Shigeki Kiyonaka
YR 2020
UL http://biorxiv.org/content/early/2020/05/22/2020.05.20.105296.abstract
AB The regulation of glutamate receptor localization is critical for development and synaptic plasticity in the central nervous system. Conventional biochemical and molecular biological approaches have been widely used to analyze glutamate receptor trafficking, especially for AMPA-type glutamate receptors (AMPARs). However, conflicting findings have been reported because of a lack of useful tools for analyzing endogenous AMPARs. Here, we develop a new method for the rapid and selective labeling of chemical probes to AMPARs by combining affinity-based protein labeling and bioorthogonal click chemistry under physiological conditions. This method allowed us to quantify AMPAR distribution and trafficking, which revealed some unique features of AMPARs, such as a long lifetime and a rapid recycling in neurons. This method was also successfully expanded to selectively label NMDA-type glutamate receptors. Thus, bioorthogonal two-step labeling may be a versatile tool for investigating the physiological and pathophysiological roles of glutamate receptors in neurons.Competing Interest StatementThe authors have declared no competing interest.