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....
| Main Authors: | , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2017-07-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/27744 |
| _version_ | 1811253010186108928 |
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| author | Sang Hak Lee Chaoyi Jin En Cai Pinghua Ge Yuji Ishitsuka Kai Wen Teng Andre A de Thomaz Duncan Nall Murat Baday Okunola Jeyifous Daniel Demonte Christopher M Dundas Sheldon Park Jary Y Delgado William N Green Paul R Selvin |
| author_facet | Sang Hak Lee Chaoyi Jin En Cai Pinghua Ge Yuji Ishitsuka Kai Wen Teng Andre A de Thomaz Duncan Nall Murat Baday Okunola Jeyifous Daniel Demonte Christopher M Dundas Sheldon Park Jary Y Delgado William N Green Paul R Selvin |
| author_sort | Sang Hak Lee |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-04-12T16:44:15Z |
| format | Article |
| id | doaj.art-671f3c4ec2474b219f8b84c8994f00b7 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-12T16:44:15Z |
| publishDate | 2017-07-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-671f3c4ec2474b219f8b84c8994f00b72022-12-22T03:24:39ZengeLife Sciences Publications LtdeLife2050-084X2017-07-01610.7554/eLife.27744Super-resolution imaging of synaptic and Extra-synaptic AMPA receptors with different-sized fluorescent probesSang Hak Lee0https://orcid.org/0000-0003-3434-076XChaoyi Jin1En Cai2Pinghua Ge3Yuji Ishitsuka4Kai Wen Teng5Andre A de Thomaz6Duncan Nall7Murat Baday8Okunola Jeyifous9Daniel Demonte10Christopher M Dundas11Sheldon Park12Jary Y Delgado13William N Green14https://orcid.org/0000-0003-2167-1391Paul R Selvin15https://orcid.org/0000-0002-3658-4218Department of Physics, Center for Biophysics, and Quantitative Biology, and Center for the Physics of Living Cells, University of Illinois, Urbana-Champaign, Champaign, United StatesDepartment of Physics, Center for Biophysics, and Quantitative Biology, and Center for the Physics of Living Cells, University of Illinois, Urbana-Champaign, Champaign, United StatesDepartment of Physics, Center for Biophysics, and Quantitative Biology, and Center for the Physics of Living Cells, University of Illinois, Urbana-Champaign, Champaign, United StatesDepartment of Physics, Center for Biophysics, and Quantitative Biology, and Center for the Physics of Living Cells, University of Illinois, Urbana-Champaign, Champaign, United StatesDepartment of Physics, Center for Biophysics, and Quantitative Biology, and Center for the Physics of Living Cells, University of Illinois, Urbana-Champaign, Champaign, United StatesDepartment of Physics, Center for Biophysics, and Quantitative Biology, and Center for the Physics of Living Cells, University of Illinois, Urbana-Champaign, Champaign, United StatesDepartment of Physics, Center for Biophysics, and Quantitative Biology, and Center for the Physics of Living Cells, University of Illinois, Urbana-Champaign, Champaign, United StatesDepartment of Physics, Center for Biophysics, and Quantitative Biology, and Center for the Physics of Living Cells, University of Illinois, Urbana-Champaign, Champaign, United StatesDepartment of Physics, Center for Biophysics, and Quantitative Biology, and Center for the Physics of Living Cells, University of Illinois, Urbana-Champaign, Champaign, United StatesDepartment of Neurobiology, University of Chicago and the Marine Biological Laboratory, Chicago, United StatesDepartment of Chemical and Biological Engineering, University at Buffalo, Buffalo, United StatesDepartment of Chemical and Biological Engineering, University at Buffalo, Buffalo, United StatesDepartment of Chemical and Biological Engineering, University at Buffalo, Buffalo, United StatesDepartment of Neurobiology, University of Chicago and the Marine Biological Laboratory, Chicago, United StatesDepartment of Neurobiology, University of Chicago and the Marine Biological Laboratory, Chicago, United StatesDepartment of Physics, Center for Biophysics, and Quantitative Biology, and Center for the Physics of Living Cells, University of Illinois, Urbana-Champaign, Champaign, United StatesPrevious 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.https://elifesciences.org/articles/27744AMAP receptorssynapsessuper-resolution imagingsmall qdotsglutamate receptorsingle molecule tracking |
| spellingShingle | Sang Hak Lee Chaoyi Jin En Cai Pinghua Ge Yuji Ishitsuka Kai Wen Teng Andre A de Thomaz Duncan Nall Murat Baday Okunola Jeyifous Daniel Demonte Christopher M Dundas Sheldon Park Jary Y Delgado William N Green Paul R Selvin Super-resolution imaging of synaptic and Extra-synaptic AMPA receptors with different-sized fluorescent probes eLife AMAP receptors synapses super-resolution imaging small qdots glutamate receptor single molecule tracking |
| 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 | AMAP receptors synapses super-resolution imaging small qdots glutamate receptor single molecule tracking |
| url | https://elifesciences.org/articles/27744 |
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