In vivo determination of fluctuating forces during endosome trafficking using a combination of active and passive microrheology.
BACKGROUND: Regulation of intracellular trafficking is a central issue in cell biology. The forces acting on intracellular vesicles (endosomes) can be assessed in living cells by using a combination of active and passive microrheology. METHODOLOGY/PRINCIPAL FINDINGS: This dual approach is based on e...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2010-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC2850365?pdf=render |
| _version_ | 1811268630661300224 |
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| author | Damien Robert Thi-Hanh Nguyen François Gallet Claire Wilhelm |
| author_facet | Damien Robert Thi-Hanh Nguyen François Gallet Claire Wilhelm |
| author_sort | Damien Robert |
| collection | DOAJ |
| description | BACKGROUND: Regulation of intracellular trafficking is a central issue in cell biology. The forces acting on intracellular vesicles (endosomes) can be assessed in living cells by using a combination of active and passive microrheology. METHODOLOGY/PRINCIPAL FINDINGS: This dual approach is based on endosome labeling with magnetic nanoparticles. The resulting magnetic endosomes act both as probes that can be manipulated with external magnetic fields to infer the viscoelastic modulus of their surrounding microenvironment, and as biological vehicles that are trafficked along the microtubule network by means of forces generated by molecular motors. The intracellular viscoelastic modulus exhibits power law dependence with frequency, which is microtubule and actin-dependent. The mean square displacements of endosomes do not follow the predictions of the fluctuation-dissipation theorem, which offers evidence for active force generation. Microtubule disruption brings the intracellular medium closer to thermal equilibrium: active forces acting on the endosomes depend on microtubule-associated motors. The power spectra of these active forces, deduced through the use of a generalized Langevin equation, show a power law decrease with frequency and reveal an actin-dependent persistence of the force with time. Experimental spectra have been reproduced by a simple model consisting in a series of force steps power-law distributed in time. This model enlightens the role of the cytoskeleton dependent force exerted on endosomes to perform intracellular trafficking. CONCLUSIONS/SIGNIFICANCE: In this work, the influence of cytoskeleton components and molecular motors on intracellular viscoelasticity and transport is addressed. The use of an original probe, the magnetic endosome, allows retrieving the power spectrum of active forces on these organelles thanks to interrelated active and passive measures. Finally a computational model gives estimates of the force itself and hence of the number of the motors pulling on endosomes. |
| first_indexed | 2024-04-12T21:25:16Z |
| format | Article |
| id | doaj.art-f3492b82972940e99f3875552a989aab |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-04-12T21:25:16Z |
| publishDate | 2010-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-f3492b82972940e99f3875552a989aab2022-12-22T03:16:11ZengPublic Library of Science (PLoS)PLoS ONE1932-62032010-01-0154e1004610.1371/journal.pone.0010046In vivo determination of fluctuating forces during endosome trafficking using a combination of active and passive microrheology.Damien RobertThi-Hanh NguyenFrançois GalletClaire WilhelmBACKGROUND: Regulation of intracellular trafficking is a central issue in cell biology. The forces acting on intracellular vesicles (endosomes) can be assessed in living cells by using a combination of active and passive microrheology. METHODOLOGY/PRINCIPAL FINDINGS: This dual approach is based on endosome labeling with magnetic nanoparticles. The resulting magnetic endosomes act both as probes that can be manipulated with external magnetic fields to infer the viscoelastic modulus of their surrounding microenvironment, and as biological vehicles that are trafficked along the microtubule network by means of forces generated by molecular motors. The intracellular viscoelastic modulus exhibits power law dependence with frequency, which is microtubule and actin-dependent. The mean square displacements of endosomes do not follow the predictions of the fluctuation-dissipation theorem, which offers evidence for active force generation. Microtubule disruption brings the intracellular medium closer to thermal equilibrium: active forces acting on the endosomes depend on microtubule-associated motors. The power spectra of these active forces, deduced through the use of a generalized Langevin equation, show a power law decrease with frequency and reveal an actin-dependent persistence of the force with time. Experimental spectra have been reproduced by a simple model consisting in a series of force steps power-law distributed in time. This model enlightens the role of the cytoskeleton dependent force exerted on endosomes to perform intracellular trafficking. CONCLUSIONS/SIGNIFICANCE: In this work, the influence of cytoskeleton components and molecular motors on intracellular viscoelasticity and transport is addressed. The use of an original probe, the magnetic endosome, allows retrieving the power spectrum of active forces on these organelles thanks to interrelated active and passive measures. Finally a computational model gives estimates of the force itself and hence of the number of the motors pulling on endosomes.http://europepmc.org/articles/PMC2850365?pdf=render |
| spellingShingle | Damien Robert Thi-Hanh Nguyen François Gallet Claire Wilhelm In vivo determination of fluctuating forces during endosome trafficking using a combination of active and passive microrheology. PLoS ONE |
| title | In vivo determination of fluctuating forces during endosome trafficking using a combination of active and passive microrheology. |
| title_full | In vivo determination of fluctuating forces during endosome trafficking using a combination of active and passive microrheology. |
| title_fullStr | In vivo determination of fluctuating forces during endosome trafficking using a combination of active and passive microrheology. |
| title_full_unstemmed | In vivo determination of fluctuating forces during endosome trafficking using a combination of active and passive microrheology. |
| title_short | In vivo determination of fluctuating forces during endosome trafficking using a combination of active and passive microrheology. |
| title_sort | in vivo determination of fluctuating forces during endosome trafficking using a combination of active and passive microrheology |
| url | http://europepmc.org/articles/PMC2850365?pdf=render |
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