Soga Kyohei, Fujiwara Takaaki, Nakagawa Mayu, Shibata Akihiro, Adriel Hansel, Yatsuzuka Kenji, Kakegawa Wataru, Yuzaki Michisuke, Hamachi Itaru, Nango Eriko, Kiyonaka Shigeki
Department of Biomolecular Engineering, Graduate School of Engineering, Nagoya University, Nagoya, Aichi 464-8603, Japan.
Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, Sendai, Miyagi 980-8577, Japan.
Sci Adv. 2025 Jun 6;11(23):eadt6683. doi: 10.1126/sciadv.adt6683.
The subcellular localization of neurotransmitter receptors is strictly regulated in neurons. Changes in the trafficking of α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-type glutamate receptors (AMPARs) play an essential role in synaptic plasticity, which is the cellular basis of learning and memory. To explore receptor trafficking, genetically encoded approaches (e.g., the fusion of fluorescent proteins to receptors) are often used. However, concerns remain that genetic approaches cannot fully reproduce the receptor functions that are inherent to neurons. Herein, we report on PFQX1(AF488), a fluorescent probe for the visualization of cell-surface AMPARs without any genetic manipulation to neurons. The rapid and reversible staining features of this probe enabled snapshot imaging, which showed the accumulation of native AMPARs in dendritic spines during synaptic plasticity. Moreover, the mechanism of this synaptic accumulation, for which genetically encoded approaches have given controversial results, was revealed by integrating two chemical methods: PFQX1(AF488) and covalent chemical labeling.
神经递质受体的亚细胞定位在神经元中受到严格调控。α-氨基-3-羟基-5-甲基-4-异恶唑丙酸（AMPA）型谷氨酸受体（AMPARs）的转运变化在突触可塑性中起着至关重要的作用，而突触可塑性是学习和记忆的细胞基础。为了探索受体转运，人们经常使用基因编码方法（例如，将荧光蛋白与受体融合）。然而，人们仍然担心基因方法无法完全重现神经元固有的受体功能。在此，我们报告了PFQX1（AF488），一种无需对神经元进行任何基因操作即可用于可视化细胞表面AMPARs的荧光探针。该探针快速且可逆的染色特性实现了快照成像，显示了在突触可塑性过程中天然AMPARs在树突棘中的积累。此外，通过整合两种化学方法：PFQX1（AF488）和共价化学标记，揭示了这种突触积累的机制，而基因编码方法在这方面给出了有争议的结果。
