Laser nanodissection for intracellular actin stress fibers and analysis of the severed fiber retraction and regeneration behavior
The intracellular actin cytoskeleton binds to non-muscle myosin II to form stress fibers (SFs) with a diameter of several hundred nanometers, which generate intracellular tension and control cell shape and movement. SFs generate a contractile force and transmit it to focal adhesions, which is essent...
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| Format: | Article |
| Language: | Japanese |
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The Japan Society of Mechanical Engineers
2025-03-01
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| Series: | Nihon Kikai Gakkai ronbunshu |
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| Online Access: | https://www.jstage.jst.go.jp/article/transjsme/91/944/91_24-00271/_pdf/-char/en |
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| _version_ | 1850282892042698752 |
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| author | Chiho KAMBE Kazuaki NAGAYAMA |
| author_facet | Chiho KAMBE Kazuaki NAGAYAMA |
| author_sort | Chiho KAMBE |
| collection | DOAJ |
| description | The intracellular actin cytoskeleton binds to non-muscle myosin II to form stress fibers (SFs) with a diameter of several hundred nanometers, which generate intracellular tension and control cell shape and movement. SFs generate a contractile force and transmit it to focal adhesions, which is essential for regulation of cell adhesion to extracellular matrix and cellular sensing of extracellular mechanical environments. Although several studies have been reported on the mechanical and contractile properties of SFs, it has not been clear how completely severed SFs are repaired and regenerated. In this study, SFs located at the periphery of living myoblasts (C2C12) were severed using a pulsed laser and their contraction and repair process were analyzed by fluorescence observation. SFs immediately contracted after their severing. The contraction process of SFs was accurately fitted using the first order lag model, indicating viscoelastic behavior. We found that SFs have a potential to repair their structures: in approximately 80% of the cells, the structure of severed SFs was gradually repaired over several minutes with contraction behavior (Repaired group), but not in some cells (Unrepaired group). The contraction rate α and contraction time constant τ of the SFs in the unrepaired group tended to be larger than those of the repaired group. These results suggest that the repair process of the severed SFs is possibly affected by the intrinsic mechanical tension of the SFs and viscous resistance from the intracellular components around the SFs which is generated by the sliding motion caused by the fiber contraction. |
| format | Article |
| id | doaj-art-cafa82176b4a4937b634c6a1df42c647 |
| institution | OA Journals |
| issn | 2187-9761 |
| language | Japanese |
| publishDate | 2025-03-01 |
| publisher | The Japan Society of Mechanical Engineers |
| record_format | Article |
| series | Nihon Kikai Gakkai ronbunshu |
| spelling | doaj-art-cafa82176b4a4937b634c6a1df42c6472025-08-20T01:47:51ZjpnThe Japan Society of Mechanical EngineersNihon Kikai Gakkai ronbunshu2187-97612025-03-019194424-0027124-0027110.1299/transjsme.24-00271transjsmeLaser nanodissection for intracellular actin stress fibers and analysis of the severed fiber retraction and regeneration behaviorChiho KAMBE0Kazuaki NAGAYAMA1Micro-Nano Biomechanics Laboratory, Department of Mechanical Systems Engineering, Ibaraki UniversityMicro-Nano Biomechanics Laboratory, Department of Mechanical Systems Engineering, Ibaraki UniversityThe intracellular actin cytoskeleton binds to non-muscle myosin II to form stress fibers (SFs) with a diameter of several hundred nanometers, which generate intracellular tension and control cell shape and movement. SFs generate a contractile force and transmit it to focal adhesions, which is essential for regulation of cell adhesion to extracellular matrix and cellular sensing of extracellular mechanical environments. Although several studies have been reported on the mechanical and contractile properties of SFs, it has not been clear how completely severed SFs are repaired and regenerated. In this study, SFs located at the periphery of living myoblasts (C2C12) were severed using a pulsed laser and their contraction and repair process were analyzed by fluorescence observation. SFs immediately contracted after their severing. The contraction process of SFs was accurately fitted using the first order lag model, indicating viscoelastic behavior. We found that SFs have a potential to repair their structures: in approximately 80% of the cells, the structure of severed SFs was gradually repaired over several minutes with contraction behavior (Repaired group), but not in some cells (Unrepaired group). The contraction rate α and contraction time constant τ of the SFs in the unrepaired group tended to be larger than those of the repaired group. These results suggest that the repair process of the severed SFs is possibly affected by the intrinsic mechanical tension of the SFs and viscous resistance from the intracellular components around the SFs which is generated by the sliding motion caused by the fiber contraction.https://www.jstage.jst.go.jp/article/transjsme/91/944/91_24-00271/_pdf/-char/encell biomechanicsmechanobiologycytoskeletonlaser ablationviscoelastic contraction |
| spellingShingle | Chiho KAMBE Kazuaki NAGAYAMA Laser nanodissection for intracellular actin stress fibers and analysis of the severed fiber retraction and regeneration behavior Nihon Kikai Gakkai ronbunshu cell biomechanics mechanobiology cytoskeleton laser ablation viscoelastic contraction |
| title | Laser nanodissection for intracellular actin stress fibers and analysis of the severed fiber retraction and regeneration behavior |
| title_full | Laser nanodissection for intracellular actin stress fibers and analysis of the severed fiber retraction and regeneration behavior |
| title_fullStr | Laser nanodissection for intracellular actin stress fibers and analysis of the severed fiber retraction and regeneration behavior |
| title_full_unstemmed | Laser nanodissection for intracellular actin stress fibers and analysis of the severed fiber retraction and regeneration behavior |
| title_short | Laser nanodissection for intracellular actin stress fibers and analysis of the severed fiber retraction and regeneration behavior |
| title_sort | laser nanodissection for intracellular actin stress fibers and analysis of the severed fiber retraction and regeneration behavior |
| topic | cell biomechanics mechanobiology cytoskeleton laser ablation viscoelastic contraction |
| url | https://www.jstage.jst.go.jp/article/transjsme/91/944/91_24-00271/_pdf/-char/en |
| work_keys_str_mv | AT chihokambe lasernanodissectionforintracellularactinstressfibersandanalysisoftheseveredfiberretractionandregenerationbehavior AT kazuakinagayama lasernanodissectionforintracellularactinstressfibersandanalysisoftheseveredfiberretractionandregenerationbehavior |