Correlative STED and Atomic Force Microscopy on Live Astrocytes Reveals Plasticity of Cytoskeletal Structure and Membrane Physical Properties during Polarized Migration
The plasticity of the cytoskeleton architecture and membrane properties is important for the establishment of cell polarity, adhesion and migration. Here, we present a method which combines stimulated emission depletion (STED) super-resolution imaging and atomic force microscopy (AFM) to correlate c...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2017-04-01
|
| Series: | Frontiers in Cellular Neuroscience |
| Subjects: | |
| Online Access: | http://journal.frontiersin.org/article/10.3389/fncel.2017.00104/full |
| _version_ | 1811280709701074944 |
|---|---|
| author | Nathalie Rouach Nathalie Rouach Clemens F. Kaminski Nathan Curry Grégory Ghézali Grégory Ghézali Grégory Ghézali Gabriele S. Kaminski Schierle |
| author_facet | Nathalie Rouach Nathalie Rouach Clemens F. Kaminski Nathan Curry Grégory Ghézali Grégory Ghézali Grégory Ghézali Gabriele S. Kaminski Schierle |
| author_sort | Nathalie Rouach |
| collection | DOAJ |
| description | The plasticity of the cytoskeleton architecture and membrane properties is important for the establishment of cell polarity, adhesion and migration. Here, we present a method which combines stimulated emission depletion (STED) super-resolution imaging and atomic force microscopy (AFM) to correlate cytoskeletal structural information with membrane physical properties in live astrocytes. Using STED compatible dyes for live cell imaging of the cytoskeleton, and simultaneously mapping the cell surface topology with AFM, we obtain unprecedented detail of highly organized networks of actin and microtubules in astrocytes. Combining mechanical data from AFM with optical imaging of actin and tubulin further reveals links between cytoskeleton organization and membrane properties. Using this methodology we illustrate that scratch-induced migration induces cytoskeleton remodeling. The latter is caused by a polarization of actin and microtubule elements within astroglial cell processes, which correlates strongly with changes in cell stiffness. The method opens new avenues for the dynamic probing of the membrane structural and functional plasticity of living brain cells. It is a powerful tool for providing new insights into mechanisms of cell structural remodeling during physiological or pathological processes, such as brain development or tumorigenesis. |
| first_indexed | 2024-04-13T01:20:48Z |
| format | Article |
| id | doaj.art-28fe22670e9b4b679c83e749bbeaad9e |
| institution | Directory Open Access Journal |
| issn | 1662-5102 |
| language | English |
| last_indexed | 2024-04-13T01:20:48Z |
| publishDate | 2017-04-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cellular Neuroscience |
| spelling | doaj.art-28fe22670e9b4b679c83e749bbeaad9e2022-12-22T03:08:47ZengFrontiers Media S.A.Frontiers in Cellular Neuroscience1662-51022017-04-011110.3389/fncel.2017.00104239497Correlative STED and Atomic Force Microscopy on Live Astrocytes Reveals Plasticity of Cytoskeletal Structure and Membrane Physical Properties during Polarized MigrationNathalie Rouach0Nathalie Rouach1Clemens F. Kaminski2Nathan Curry3Grégory Ghézali4Grégory Ghézali5Grégory Ghézali6Gabriele S. Kaminski Schierle7Chemical Engineering and Biotechnology, University of CambridgeCambridge, UKCenter for Interdisciplinary Research in Biology, College de France, CNRS UMR 7241, INSERM U1050, Labex Memolife, PSL Research UniversityParis, FranceChemical Engineering and Biotechnology, University of CambridgeCambridge, UKChemical Engineering and Biotechnology, University of CambridgeCambridge, UKChemical Engineering and Biotechnology, University of CambridgeCambridge, UKCenter for Interdisciplinary Research in Biology, College de France, CNRS UMR 7241, INSERM U1050, Labex Memolife, PSL Research UniversityParis, FranceDoctoral School No 158, Pierre and Marie Curie UniversityParis, FranceChemical Engineering and Biotechnology, University of CambridgeCambridge, UKThe plasticity of the cytoskeleton architecture and membrane properties is important for the establishment of cell polarity, adhesion and migration. Here, we present a method which combines stimulated emission depletion (STED) super-resolution imaging and atomic force microscopy (AFM) to correlate cytoskeletal structural information with membrane physical properties in live astrocytes. Using STED compatible dyes for live cell imaging of the cytoskeleton, and simultaneously mapping the cell surface topology with AFM, we obtain unprecedented detail of highly organized networks of actin and microtubules in astrocytes. Combining mechanical data from AFM with optical imaging of actin and tubulin further reveals links between cytoskeleton organization and membrane properties. Using this methodology we illustrate that scratch-induced migration induces cytoskeleton remodeling. The latter is caused by a polarization of actin and microtubule elements within astroglial cell processes, which correlates strongly with changes in cell stiffness. The method opens new avenues for the dynamic probing of the membrane structural and functional plasticity of living brain cells. It is a powerful tool for providing new insights into mechanisms of cell structural remodeling during physiological or pathological processes, such as brain development or tumorigenesis.http://journal.frontiersin.org/article/10.3389/fncel.2017.00104/fullastrocytesmigrationprotrusionsmembrane physical propertiescytoskeletonatomic force microscopy |
| spellingShingle | Nathalie Rouach Nathalie Rouach Clemens F. Kaminski Nathan Curry Grégory Ghézali Grégory Ghézali Grégory Ghézali Gabriele S. Kaminski Schierle Correlative STED and Atomic Force Microscopy on Live Astrocytes Reveals Plasticity of Cytoskeletal Structure and Membrane Physical Properties during Polarized Migration Frontiers in Cellular Neuroscience astrocytes migration protrusions membrane physical properties cytoskeleton atomic force microscopy |
| title | Correlative STED and Atomic Force Microscopy on Live Astrocytes Reveals Plasticity of Cytoskeletal Structure and Membrane Physical Properties during Polarized Migration |
| title_full | Correlative STED and Atomic Force Microscopy on Live Astrocytes Reveals Plasticity of Cytoskeletal Structure and Membrane Physical Properties during Polarized Migration |
| title_fullStr | Correlative STED and Atomic Force Microscopy on Live Astrocytes Reveals Plasticity of Cytoskeletal Structure and Membrane Physical Properties during Polarized Migration |
| title_full_unstemmed | Correlative STED and Atomic Force Microscopy on Live Astrocytes Reveals Plasticity of Cytoskeletal Structure and Membrane Physical Properties during Polarized Migration |
| title_short | Correlative STED and Atomic Force Microscopy on Live Astrocytes Reveals Plasticity of Cytoskeletal Structure and Membrane Physical Properties during Polarized Migration |
| title_sort | correlative sted and atomic force microscopy on live astrocytes reveals plasticity of cytoskeletal structure and membrane physical properties during polarized migration |
| topic | astrocytes migration protrusions membrane physical properties cytoskeleton atomic force microscopy |
| url | http://journal.frontiersin.org/article/10.3389/fncel.2017.00104/full |
| work_keys_str_mv | AT nathalierouach correlativestedandatomicforcemicroscopyonliveastrocytesrevealsplasticityofcytoskeletalstructureandmembranephysicalpropertiesduringpolarizedmigration AT nathalierouach correlativestedandatomicforcemicroscopyonliveastrocytesrevealsplasticityofcytoskeletalstructureandmembranephysicalpropertiesduringpolarizedmigration AT clemensfkaminski correlativestedandatomicforcemicroscopyonliveastrocytesrevealsplasticityofcytoskeletalstructureandmembranephysicalpropertiesduringpolarizedmigration AT nathancurry correlativestedandatomicforcemicroscopyonliveastrocytesrevealsplasticityofcytoskeletalstructureandmembranephysicalpropertiesduringpolarizedmigration AT gregoryghezali correlativestedandatomicforcemicroscopyonliveastrocytesrevealsplasticityofcytoskeletalstructureandmembranephysicalpropertiesduringpolarizedmigration AT gregoryghezali correlativestedandatomicforcemicroscopyonliveastrocytesrevealsplasticityofcytoskeletalstructureandmembranephysicalpropertiesduringpolarizedmigration AT gregoryghezali correlativestedandatomicforcemicroscopyonliveastrocytesrevealsplasticityofcytoskeletalstructureandmembranephysicalpropertiesduringpolarizedmigration AT gabrieleskaminskischierle correlativestedandatomicforcemicroscopyonliveastrocytesrevealsplasticityofcytoskeletalstructureandmembranephysicalpropertiesduringpolarizedmigration |