Characterization of microtubule buckling in living cells
Microtubules are filamentous biopolymers involved in essential biological processes. They form key structures in eukaryotic cells, and thus it is very important to determine the mechanisms involved in the formation and maintenance of the microtubule network. Microtubule bucklings are transient and l...
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2017
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| Acceso en línea: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01757571_v46_n6_p581_Pallavicini http://hdl.handle.net/20.500.12110/paper_01757571_v46_n6_p581_Pallavicini |
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paper:paper_01757571_v46_n6_p581_Pallavicini2023-06-08T15:18:56Z Characterization of microtubule buckling in living cells Wetzler, Diana E. Levi, Valeria Bruno, Luciana Active forces Buckling Filament tracking Fluorescence microscopy Living cells Microtubules molecular motor animal cell Article cell organelle confocal microscopy controlled study cytoskeleton endosome fluorescence analysis fluorescence microscopy intracellular space melanophore microfilament microtubule nonhuman polymerization priority journal simulation Xenopus laevis animal biological model biomechanics cell line cell survival mechanics metabolism microtubule movement (physiology) Animals Biomechanical Phenomena Cell Line Cell Survival Mechanical Phenomena Microtubules Models, Biological Movement Xenopus laevis Microtubules are filamentous biopolymers involved in essential biological processes. They form key structures in eukaryotic cells, and thus it is very important to determine the mechanisms involved in the formation and maintenance of the microtubule network. Microtubule bucklings are transient and localized events commonly observed in living cells and characterized by a fast bending and its posterior relaxation. Active forces provided by molecular motors have been indicated as responsible for most of these rapid deformations. However, the factors that control the shape amplitude and the time scales of the rising and release stages remain unexplored. In this work, we study microtubule buckling in living cells using Xenopus laevis melanophores as a model system. We tracked single fluorescent microtubules from high temporal resolution (0.3–2 s) confocal movies. We recovered the center coordinates of the filaments with 10-nm precision and analyzed the amplitude of the deformation as a function of time. Using numerical simulations, we explored different force mechanisms resulting in microtubule bending. The simulated events reproduce many features observed for microtubules, suggesting that a mechanistic model captures the essential processes underlying microtubule buckling. Also, we studied the interplay between actively transported vesicles and the microtubule network using a two-color technique. Our results suggest that microtubules may affect transport indirectly besides serving as tracks of motor-driven organelles. For example, they could obstruct organelles at microtubule intersections or push them during filament mechanical relaxation. © 2017, European Biophysical Societies' Association. Fil:Wetzler, D. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Levi, V. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Bruno, L. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2017 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01757571_v46_n6_p581_Pallavicini http://hdl.handle.net/20.500.12110/paper_01757571_v46_n6_p581_Pallavicini |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Active forces Buckling Filament tracking Fluorescence microscopy Living cells Microtubules molecular motor animal cell Article cell organelle confocal microscopy controlled study cytoskeleton endosome fluorescence analysis fluorescence microscopy intracellular space melanophore microfilament microtubule nonhuman polymerization priority journal simulation Xenopus laevis animal biological model biomechanics cell line cell survival mechanics metabolism microtubule movement (physiology) Animals Biomechanical Phenomena Cell Line Cell Survival Mechanical Phenomena Microtubules Models, Biological Movement Xenopus laevis |
| spellingShingle |
Active forces Buckling Filament tracking Fluorescence microscopy Living cells Microtubules molecular motor animal cell Article cell organelle confocal microscopy controlled study cytoskeleton endosome fluorescence analysis fluorescence microscopy intracellular space melanophore microfilament microtubule nonhuman polymerization priority journal simulation Xenopus laevis animal biological model biomechanics cell line cell survival mechanics metabolism microtubule movement (physiology) Animals Biomechanical Phenomena Cell Line Cell Survival Mechanical Phenomena Microtubules Models, Biological Movement Xenopus laevis Wetzler, Diana E. Levi, Valeria Bruno, Luciana Characterization of microtubule buckling in living cells |
| topic_facet |
Active forces Buckling Filament tracking Fluorescence microscopy Living cells Microtubules molecular motor animal cell Article cell organelle confocal microscopy controlled study cytoskeleton endosome fluorescence analysis fluorescence microscopy intracellular space melanophore microfilament microtubule nonhuman polymerization priority journal simulation Xenopus laevis animal biological model biomechanics cell line cell survival mechanics metabolism microtubule movement (physiology) Animals Biomechanical Phenomena Cell Line Cell Survival Mechanical Phenomena Microtubules Models, Biological Movement Xenopus laevis |
| description |
Microtubules are filamentous biopolymers involved in essential biological processes. They form key structures in eukaryotic cells, and thus it is very important to determine the mechanisms involved in the formation and maintenance of the microtubule network. Microtubule bucklings are transient and localized events commonly observed in living cells and characterized by a fast bending and its posterior relaxation. Active forces provided by molecular motors have been indicated as responsible for most of these rapid deformations. However, the factors that control the shape amplitude and the time scales of the rising and release stages remain unexplored. In this work, we study microtubule buckling in living cells using Xenopus laevis melanophores as a model system. We tracked single fluorescent microtubules from high temporal resolution (0.3–2 s) confocal movies. We recovered the center coordinates of the filaments with 10-nm precision and analyzed the amplitude of the deformation as a function of time. Using numerical simulations, we explored different force mechanisms resulting in microtubule bending. The simulated events reproduce many features observed for microtubules, suggesting that a mechanistic model captures the essential processes underlying microtubule buckling. Also, we studied the interplay between actively transported vesicles and the microtubule network using a two-color technique. Our results suggest that microtubules may affect transport indirectly besides serving as tracks of motor-driven organelles. For example, they could obstruct organelles at microtubule intersections or push them during filament mechanical relaxation. © 2017, European Biophysical Societies' Association. |
| author |
Wetzler, Diana E. Levi, Valeria Bruno, Luciana |
| author_facet |
Wetzler, Diana E. Levi, Valeria Bruno, Luciana |
| author_sort |
Wetzler, Diana E. |
| title |
Characterization of microtubule buckling in living cells |
| title_short |
Characterization of microtubule buckling in living cells |
| title_full |
Characterization of microtubule buckling in living cells |
| title_fullStr |
Characterization of microtubule buckling in living cells |
| title_full_unstemmed |
Characterization of microtubule buckling in living cells |
| title_sort |
characterization of microtubule buckling in living cells |
| publishDate |
2017 |
| url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01757571_v46_n6_p581_Pallavicini http://hdl.handle.net/20.500.12110/paper_01757571_v46_n6_p581_Pallavicini |
| work_keys_str_mv |
AT wetzlerdianae characterizationofmicrotubulebucklinginlivingcells AT levivaleria characterizationofmicrotubulebucklinginlivingcells AT brunoluciana characterizationofmicrotubulebucklinginlivingcells |
| _version_ |
1768544953781190656 |