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Sci Rep. 2015 Jun 29;5:11454. doi: 10.1038/srep11454.

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

Author information

1
1] Department of Nanobiophotonics, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, Göttingen 37077, Germany [2] Centre for Neural Circuits and Behaviour, University of Oxford, Mansfield Road, Oxford OX1 3SR, UK.
2
1] Department of Nanobiophotonics, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, Göttingen 37077, Germany [2] MRC Human Immunology Unit and Wolfson Imaging Centre Oxford, Weatherall Institute of Molecular Medicine, University of Oxford, Headley Way, Oxford OX3 9DS, UK [3] MEMPHYS - Center for Biomembrane Physics, University of Southern Denmark, Campusvej 55, Odense M,DK-5230, Denmark.
3
Department of Nanobiophotonics, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, Göttingen 37077, Germany.
4
Department of Cell &Developmental Biology, and Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill NC 27599, USA.
5
1] Department of Cell &Developmental Biology, and Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill NC 27599, USA [2] Howard Hughes Medical Institute, University of North Carolina, Chapel Hill NC 27599, USA.
6
1] MRC Human Immunology Unit and Wolfson Imaging Centre Oxford, Weatherall Institute of Molecular Medicine, University of Oxford, Headley Way, Oxford OX3 9DS, UK [2] MEMPHYS - Center for Biomembrane Physics, University of Southern Denmark, Campusvej 55, Odense M,DK-5230, Denmark.
7
1] Department of Nanobiophotonics, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, Göttingen 37077, Germany [2] MRC Human Immunology Unit and Wolfson Imaging Centre Oxford, Weatherall Institute of Molecular Medicine, University of Oxford, Headley Way, Oxford OX3 9DS, UK.

Abstract

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.

PMID:
26118385
PMCID:
PMC4484492
DOI:
10.1038/srep11454
[Indexed for MEDLINE]
Free PMC Article

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