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Super-resolution imaging of synaptic and Extra-synaptic AMPA receptors with different-sized fluorescent probes

Previous studies tracking AMPA receptor (AMPAR) diffusion at synapses observed a large mobile extrasynaptic AMPAR pool. Using super-resolution microscopy, we examined how fluorophore size and photostability affected AMPAR trafficking outside of, and within, post-synaptic densities (PSDs) from rats....

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Autores principales: Lee, Sang Hak, Jin, Chaoyi, Cai, En, Ge, Pinghua, Ishitsuka, Yuji, Teng, Kai Wen, de Thomaz, Andre A, Nall, Duncan, Baday, Murat, Jeyifous, Okunola, Demonte, Daniel, Dundas, Christopher M, Park, Sheldon, Delgado, Jary Y, Green, William N, Selvin, Paul R
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5779237/
https://www.ncbi.nlm.nih.gov/pubmed/28749340
http://dx.doi.org/10.7554/eLife.27744
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author Lee, Sang Hak
Jin, Chaoyi
Cai, En
Ge, Pinghua
Ishitsuka, Yuji
Teng, Kai Wen
de Thomaz, Andre A
Nall, Duncan
Baday, Murat
Jeyifous, Okunola
Demonte, Daniel
Dundas, Christopher M
Park, Sheldon
Delgado, Jary Y
Green, William N
Selvin, Paul R
author_facet Lee, Sang Hak
Jin, Chaoyi
Cai, En
Ge, Pinghua
Ishitsuka, Yuji
Teng, Kai Wen
de Thomaz, Andre A
Nall, Duncan
Baday, Murat
Jeyifous, Okunola
Demonte, Daniel
Dundas, Christopher M
Park, Sheldon
Delgado, Jary Y
Green, William N
Selvin, Paul R
author_sort Lee, Sang Hak
collection PubMed
description Previous studies tracking AMPA receptor (AMPAR) diffusion at synapses observed a large mobile extrasynaptic AMPAR pool. Using super-resolution microscopy, we examined how fluorophore size and photostability affected AMPAR trafficking outside of, and within, post-synaptic densities (PSDs) from rats. Organic fluorescent dyes (≈4 nm), quantum dots, either small (≈10 nm diameter; sQDs) or big (>20 nm; bQDs), were coupled to AMPARs via different-sized linkers. We find that >90% of AMPARs labeled with fluorescent dyes or sQDs were diffusing in confined nanodomains in PSDs, which were stable for 15 min or longer. Less than 10% of sQD-AMPARs were extrasynaptic and highly mobile. In contrast, 5–10% of bQD-AMPARs were in PSDs and 90–95% were extrasynaptic as previously observed. Contrary to the hypothesis that AMPAR entry is limited by the occupancy of open PSD ‘slots’, our findings suggest that AMPARs rapidly enter stable ‘nanodomains’ in PSDs with lifetime >15 min, and do not accumulate in extrasynaptic membranes.
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spelling pubmed-57792372018-01-25 Super-resolution imaging of synaptic and Extra-synaptic AMPA receptors with different-sized fluorescent probes Lee, Sang Hak Jin, Chaoyi Cai, En Ge, Pinghua Ishitsuka, Yuji Teng, Kai Wen de Thomaz, Andre A Nall, Duncan Baday, Murat Jeyifous, Okunola Demonte, Daniel Dundas, Christopher M Park, Sheldon Delgado, Jary Y Green, William N Selvin, Paul R eLife Structural Biology and Molecular Biophysics Previous studies tracking AMPA receptor (AMPAR) diffusion at synapses observed a large mobile extrasynaptic AMPAR pool. Using super-resolution microscopy, we examined how fluorophore size and photostability affected AMPAR trafficking outside of, and within, post-synaptic densities (PSDs) from rats. Organic fluorescent dyes (≈4 nm), quantum dots, either small (≈10 nm diameter; sQDs) or big (>20 nm; bQDs), were coupled to AMPARs via different-sized linkers. We find that >90% of AMPARs labeled with fluorescent dyes or sQDs were diffusing in confined nanodomains in PSDs, which were stable for 15 min or longer. Less than 10% of sQD-AMPARs were extrasynaptic and highly mobile. In contrast, 5–10% of bQD-AMPARs were in PSDs and 90–95% were extrasynaptic as previously observed. Contrary to the hypothesis that AMPAR entry is limited by the occupancy of open PSD ‘slots’, our findings suggest that AMPARs rapidly enter stable ‘nanodomains’ in PSDs with lifetime >15 min, and do not accumulate in extrasynaptic membranes. eLife Sciences Publications, Ltd 2017-07-27 /pmc/articles/PMC5779237/ /pubmed/28749340 http://dx.doi.org/10.7554/eLife.27744 Text en © 2017, Lee et al http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Structural Biology and Molecular Biophysics
Lee, Sang Hak
Jin, Chaoyi
Cai, En
Ge, Pinghua
Ishitsuka, Yuji
Teng, Kai Wen
de Thomaz, Andre A
Nall, Duncan
Baday, Murat
Jeyifous, Okunola
Demonte, Daniel
Dundas, Christopher M
Park, Sheldon
Delgado, Jary Y
Green, William N
Selvin, Paul R
Super-resolution imaging of synaptic and Extra-synaptic AMPA receptors with different-sized fluorescent probes
title Super-resolution imaging of synaptic and Extra-synaptic AMPA receptors with different-sized fluorescent probes
title_full Super-resolution imaging of synaptic and Extra-synaptic AMPA receptors with different-sized fluorescent probes
title_fullStr Super-resolution imaging of synaptic and Extra-synaptic AMPA receptors with different-sized fluorescent probes
title_full_unstemmed Super-resolution imaging of synaptic and Extra-synaptic AMPA receptors with different-sized fluorescent probes
title_short Super-resolution imaging of synaptic and Extra-synaptic AMPA receptors with different-sized fluorescent probes
title_sort super-resolution imaging of synaptic and extra-synaptic ampa receptors with different-sized fluorescent probes
topic Structural Biology and Molecular Biophysics
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5779237/
https://www.ncbi.nlm.nih.gov/pubmed/28749340
http://dx.doi.org/10.7554/eLife.27744
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