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.

中文翻译：

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配体导向的两步标记可量化神经元谷氨酸受体的运输

谷氨酸受体定位的调节对于中枢神经系统的发育和突触可塑性至关重要。传统的生物化学和分子生物学方法已被广泛用于分析谷氨酸受体的运输，特别是对于AMPA型谷氨酸受体（AMPAR）。但是，由于缺乏用于分析内源AMPAR的有用工具，因此报告了相互矛盾的发现。在这里，我们通过结合基于亲和力的蛋白质标记和生物正交点击化学在生理条件下，开发了一种新的方法，用于对AMPARs的化学探针进行快速和选择性标记。这种方法使我们能够量化AMPAR的分布和运输，从而揭示了AMPAR的一些独特功能，例如使用寿命长和神经元快速回收。此方法也成功地扩展到选择性标记NMDA型谷氨酸受体。因此，正交的两步标记法可能是研究神经元中谷氨酸受体的生理和病理生理作用的通用工具。