Super-resolution dynamic imaging of dendritic spines using a low-affinity photoconvertible actin probe.

The actin cytoskeleton of dendritic spines plays a key role in morphological aspects of synaptic plasticity. The detailed analysis of the spine structure and dynamics in live neurons, however, has been hampered by the diffraction-limited resolution of conventional fluorescence microscopy. The advent...

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Autores principales:	Ignacio Izeddin, Christian G Specht, Mickaël Lelek, Xavier Darzacq, Antoine Triller, Christophe Zimmer, Maxime Dahan
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2011
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/a9eed8101df144108ef02e4c776a39ed
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spelling	oai:doaj.org-article:a9eed8101df144108ef02e4c776a39ed2021-11-18T07:00:24ZSuper-resolution dynamic imaging of dendritic spines using a low-affinity photoconvertible actin probe.1932-620310.1371/journal.pone.0015611https://doaj.org/article/a9eed8101df144108ef02e4c776a39ed2011-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21264214/?tool=EBIhttps://doaj.org/toc/1932-6203The actin cytoskeleton of dendritic spines plays a key role in morphological aspects of synaptic plasticity. The detailed analysis of the spine structure and dynamics in live neurons, however, has been hampered by the diffraction-limited resolution of conventional fluorescence microscopy. The advent of nanoscopic imaging techniques thus holds great promise for the study of these processes. We implemented a strategy for the visualization of morphological changes of dendritic spines over tens of minutes at a lateral resolution of 25 to 65 nm. We have generated a low-affinity photoconvertible probe, capable of reversibly binding to actin and thus allowing long-term photoactivated localization microscopy of the spine cytoskeleton. Using this approach, we resolve structural parameters of spines and record their long-term dynamics at a temporal resolution below one minute. Furthermore, we have determined changes in the spine morphology in response to pharmacologically induced synaptic activity and quantified the actin redistribution underlying these changes. By combining PALM imaging with quantum dot tracking, we could also simultaneously visualize the cytoskeleton and the spine membrane, allowing us to record complementary information on the morphological changes of the spines at super-resolution.Ignacio IzeddinChristian G SpechtMickaël LelekXavier DarzacqAntoine TrillerChristophe ZimmerMaxime DahanPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 6, Iss 1, p e15611 (2011)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Ignacio Izeddin Christian G Specht Mickaël Lelek Xavier Darzacq Antoine Triller Christophe Zimmer Maxime Dahan Super-resolution dynamic imaging of dendritic spines using a low-affinity photoconvertible actin probe.
description	The actin cytoskeleton of dendritic spines plays a key role in morphological aspects of synaptic plasticity. The detailed analysis of the spine structure and dynamics in live neurons, however, has been hampered by the diffraction-limited resolution of conventional fluorescence microscopy. The advent of nanoscopic imaging techniques thus holds great promise for the study of these processes. We implemented a strategy for the visualization of morphological changes of dendritic spines over tens of minutes at a lateral resolution of 25 to 65 nm. We have generated a low-affinity photoconvertible probe, capable of reversibly binding to actin and thus allowing long-term photoactivated localization microscopy of the spine cytoskeleton. Using this approach, we resolve structural parameters of spines and record their long-term dynamics at a temporal resolution below one minute. Furthermore, we have determined changes in the spine morphology in response to pharmacologically induced synaptic activity and quantified the actin redistribution underlying these changes. By combining PALM imaging with quantum dot tracking, we could also simultaneously visualize the cytoskeleton and the spine membrane, allowing us to record complementary information on the morphological changes of the spines at super-resolution.
format	article
author	Ignacio Izeddin Christian G Specht Mickaël Lelek Xavier Darzacq Antoine Triller Christophe Zimmer Maxime Dahan
author_facet	Ignacio Izeddin Christian G Specht Mickaël Lelek Xavier Darzacq Antoine Triller Christophe Zimmer Maxime Dahan
author_sort	Ignacio Izeddin
title	Super-resolution dynamic imaging of dendritic spines using a low-affinity photoconvertible actin probe.
title_short	Super-resolution dynamic imaging of dendritic spines using a low-affinity photoconvertible actin probe.
title_full	Super-resolution dynamic imaging of dendritic spines using a low-affinity photoconvertible actin probe.
title_fullStr	Super-resolution dynamic imaging of dendritic spines using a low-affinity photoconvertible actin probe.
title_full_unstemmed	Super-resolution dynamic imaging of dendritic spines using a low-affinity photoconvertible actin probe.
title_sort	super-resolution dynamic imaging of dendritic spines using a low-affinity photoconvertible actin probe.
publisher	Public Library of Science (PLoS)
publishDate	2011
url	https://doaj.org/article/a9eed8101df144108ef02e4c776a39ed
work_keys_str_mv	AT ignacioizeddin superresolutiondynamicimagingofdendriticspinesusingalowaffinityphotoconvertibleactinprobe AT christiangspecht superresolutiondynamicimagingofdendriticspinesusingalowaffinityphotoconvertibleactinprobe AT mickaellelek superresolutiondynamicimagingofdendriticspinesusingalowaffinityphotoconvertibleactinprobe AT xavierdarzacq superresolutiondynamicimagingofdendriticspinesusingalowaffinityphotoconvertibleactinprobe AT antoinetriller superresolutiondynamicimagingofdendriticspinesusingalowaffinityphotoconvertibleactinprobe AT christophezimmer superresolutiondynamicimagingofdendriticspinesusingalowaffinityphotoconvertibleactinprobe AT maximedahan superresolutiondynamicimagingofdendriticspinesusingalowaffinityphotoconvertibleactinprobe
_version_	1718424062407999488

Super-resolution dynamic imaging of dendritic spines using a low-affinity photoconvertible actin probe.

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