Nature communications, 2021-02, Vol.12 (1), p.831-831, Article 831
2021
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- Autor(en) / Beteiligte
- Titel
- Ligand-directed two-step labeling to quantify neuronal glutamate receptor trafficking
- Ist Teil von
- Nature communications, 2021-02, Vol.12 (1), p.831-831, Article 831
- Ort / Verlag
- London: Nature Publishing Group UK
- Erscheinungsjahr
- 2021
- Quelle
- Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals
- Beschreibungen/Notizen
- 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 α-amino-3-hydroxy-5-methyl-4-isoxazole-propionate-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 method for the rapid and selective labeling of AMPARs with chemical probes, by combining affinity-based protein labeling and bioorthogonal click chemistry under physiological temperature in culture medium. This method allows us to quantify AMPAR distribution and trafficking, which reveals some unique features of AMPARs, such as a long lifetime and a rapid recycling in neurons. This method is also successfully expanded to selectively label N -methyl-D-aspartate-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. The analysis of AMPA-type glutamate receptor (AMPAR) trafficking is essential for understanding molecular mechanisms of learning and memory, but the analytical tools are currently limited. Here, the authors report a method that combines affinity-based receptor labeling and bioorthogonal click chemistry to quantify AMPAR distribution and trafficking under physiological conditions.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 2041-1723eISSN: 2041-1723DOI: 10.1038/s41467-021-21082-xTitel-ID: cdi_doaj_primary_oai_doaj_org_article_738bc8a87bde4b819f6d2abb0650a08c
- Format
- –
- Schlagworte
- 14, 14/34, 631/443/376, 639/638/45/2783, 639/638/92/269/1149, 639/638/92/96, Affinity, Animals, Central nervous system, Cerebral Cortex - cytology, Cerebral Cortex - metabolism, Chemical synthesis, Developmental plasticity, Excitatory Amino Acid Antagonists - chemistry, Fluorescein - chemistry, Fluorescent Dyes - chemistry, Gene Expression, Glutamate receptors, Half-Life, HEK293 Cells, Hippocampus - cytology, Hippocampus - metabolism, Humanities and Social Sciences, Humans, Labeling, Ligands, Localization, Mice, Mice, Inbred ICR, Molecular modelling, multidisciplinary, N-Methyl-D-aspartic acid receptors, Neurons, Neurons - metabolism, Neurons - ultrastructure, Physiology, Primary Cell Culture, Protein Transport, Quinoxalines - chemistry, Rats, Rats, Sprague-Dawley, Receptors, Receptors, AMPA - chemistry, Receptors, AMPA - genetics, Receptors, AMPA - metabolism, Receptors, N-Methyl-D-Aspartate - chemistry, Receptors, N-Methyl-D-Aspartate - genetics, Receptors, N-Methyl-D-Aspartate - metabolism, Science, Science (multidisciplinary), Staining and Labeling - methods, Synaptic plasticity, Synaptogenesis, α-Amino-3-hydroxy-5-methyl-4-isoxazole propionic acid, α-Amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptors