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Cortical actin networks induce spatio-temporal confinement of phospholipids in the plasma membrane – a minimally invasive investigation by STED-FCS

Important discoveries in the last decades have changed our view of the plasma membrane organisation. Specifically, the cortical cytoskeleton has emerged as a key modulator of the lateral diffusion of membrane proteins. Cytoskeleton-dependent compartmentalised lipid diffusion has been proposed, but t...

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Autores principales: Andrade, Débora M., Clausen, Mathias P., Keller, Jan, Mueller, Veronika, Wu, Congying, Bear, James E., Hell, Stefan W., Lagerholm, B. Christoffer, Eggeling, Christian
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4484492/
https://www.ncbi.nlm.nih.gov/pubmed/26118385
http://dx.doi.org/10.1038/srep11454
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author Andrade, Débora M.
Clausen, Mathias P.
Keller, Jan
Mueller, Veronika
Wu, Congying
Bear, James E.
Hell, Stefan W.
Lagerholm, B. Christoffer
Eggeling, Christian
author_facet Andrade, Débora M.
Clausen, Mathias P.
Keller, Jan
Mueller, Veronika
Wu, Congying
Bear, James E.
Hell, Stefan W.
Lagerholm, B. Christoffer
Eggeling, Christian
author_sort Andrade, Débora M.
collection PubMed
description Important discoveries in the last decades have changed our view of the plasma membrane organisation. Specifically, the cortical cytoskeleton has emerged as a key modulator of the lateral diffusion of membrane proteins. Cytoskeleton-dependent compartmentalised lipid diffusion has been proposed, but this concept remains controversial because this phenomenon has thus far only been observed with artefact-prone probes in combination with a single technique: single particle tracking. In this paper, we report the first direct observation of compartmentalised phospholipid diffusion in the plasma membrane of living cells using a minimally invasive, fluorescent dye labelled lipid analogue. These observations were made using optical STED nanoscopy in combination with fluorescence correlation spectroscopy (STED-FCS), a technique which allows the study of membrane dynamics on a sub-millisecond time-scale and with a spatial resolution of down to 40 nm. Specifically, we find that compartmentalised phospholipid diffusion depends on the cortical actin cytoskeleton, and that this constrained diffusion is directly dependent on the F-actin branching nucleator Arp2/3. These findings provide solid evidence that the Arp2/3-dependent cortical actin cytoskeleton plays a pivotal role in the dynamic organisation of the plasma membrane, potentially regulating fundamental cellular processes.
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spelling pubmed-44844922015-07-08 Cortical actin networks induce spatio-temporal confinement of phospholipids in the plasma membrane – a minimally invasive investigation by STED-FCS Andrade, Débora M. Clausen, Mathias P. Keller, Jan Mueller, Veronika Wu, Congying Bear, James E. Hell, Stefan W. Lagerholm, B. Christoffer Eggeling, Christian Sci Rep Article Important discoveries in the last decades have changed our view of the plasma membrane organisation. Specifically, the cortical cytoskeleton has emerged as a key modulator of the lateral diffusion of membrane proteins. Cytoskeleton-dependent compartmentalised lipid diffusion has been proposed, but this concept remains controversial because this phenomenon has thus far only been observed with artefact-prone probes in combination with a single technique: single particle tracking. In this paper, we report the first direct observation of compartmentalised phospholipid diffusion in the plasma membrane of living cells using a minimally invasive, fluorescent dye labelled lipid analogue. These observations were made using optical STED nanoscopy in combination with fluorescence correlation spectroscopy (STED-FCS), a technique which allows the study of membrane dynamics on a sub-millisecond time-scale and with a spatial resolution of down to 40 nm. Specifically, we find that compartmentalised phospholipid diffusion depends on the cortical actin cytoskeleton, and that this constrained diffusion is directly dependent on the F-actin branching nucleator Arp2/3. These findings provide solid evidence that the Arp2/3-dependent cortical actin cytoskeleton plays a pivotal role in the dynamic organisation of the plasma membrane, potentially regulating fundamental cellular processes. Nature Publishing Group 2015-06-29 /pmc/articles/PMC4484492/ /pubmed/26118385 http://dx.doi.org/10.1038/srep11454 Text en Copyright © 2015, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Andrade, Débora M.
Clausen, Mathias P.
Keller, Jan
Mueller, Veronika
Wu, Congying
Bear, James E.
Hell, Stefan W.
Lagerholm, B. Christoffer
Eggeling, Christian
Cortical actin networks induce spatio-temporal confinement of phospholipids in the plasma membrane – a minimally invasive investigation by STED-FCS
title Cortical actin networks induce spatio-temporal confinement of phospholipids in the plasma membrane – a minimally invasive investigation by STED-FCS
title_full Cortical actin networks induce spatio-temporal confinement of phospholipids in the plasma membrane – a minimally invasive investigation by STED-FCS
title_fullStr Cortical actin networks induce spatio-temporal confinement of phospholipids in the plasma membrane – a minimally invasive investigation by STED-FCS
title_full_unstemmed Cortical actin networks induce spatio-temporal confinement of phospholipids in the plasma membrane – a minimally invasive investigation by STED-FCS
title_short Cortical actin networks induce spatio-temporal confinement of phospholipids in the plasma membrane – a minimally invasive investigation by STED-FCS
title_sort cortical actin networks induce spatio-temporal confinement of phospholipids in the plasma membrane – a minimally invasive investigation by sted-fcs
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4484492/
https://www.ncbi.nlm.nih.gov/pubmed/26118385
http://dx.doi.org/10.1038/srep11454
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