Detail of Fig5D_Video5_ctrl
Project
Title
Time-lapse images of actin-GFP FRAP on an stress fibers in non-target C2C12 cells
Description
Fluorescence recovery after photobleaching (FRAP) experiments of non-target C2C12 cells. Stress fibers in C2C12 cells expressing GFP-tagged actin were locally photobleached with 405 nm laser for 0.5 sec with 100 percent of the laser power (the size of bleached area was 1.6 x 3.6 mm), and fluo- rescence images were acquired every 0.5 sec for 1 min with 3-10 percent of the laser power.
Release, Updated
2025-04-10
License
CC BY
Kind
Image data
File Formats
.nd2
Data size
16.0 MB
Datatype
-
Molecular Function (MF)
actin filament binding
Biological Process (BP)
actin nucleation
Cellular Component (CC)
cytoskeleton
stress fiber
Biological Imaging Method
X scale
0.224 micrometer/pixel
Y scale
0.224 micrometer/pixel
Z scale
-
T scale
0.5 seconds
Image Acquisition
Experiment type
-
Microscope type
-
Acquisition mode
-
Contrast method
-
Microscope model
-
Detector model
-
Objective model
-
Filter set
-
Summary of Methods
Katsuta H, Okuda S, Nagayama K, Machiyama H, Kidoaki S, Kato M, Sokabe M, Miyata T, Hirata H. Actin crosslinking by alpha-actinin averts viscous dissipation of myosin force transmission in stress fibers. iScience. 2023 Feb1;26(3):106090.
Related paper(s)
Hiroki Katsuta, Satoru Okuda, Kazuaki Nagayama, Hiroaki Machiyama, Satoru Kidoaki, Masashi Kato, Masahiro Sokabe, Takaki Miyata, Hiroaki Hirata (2023) Actin crosslinking by alpha-actinin averts viscous dissipation of myosin force transmission in stress fibers., iScience, Volume 26, Number 3, pp. 106090
Published in 2023 Mar 17 (Electronic publication in Feb. 1, 2023, midnight )
- doi: 10.1016/j.isci.2023.106090
-
pmid: 36852278
(Abstract) Contractile force generated in actomyosin stress fibers (SFs) is transmitted along SFs to the extracellular matrix (ECM), which contributes to cell migration and sensing of ECM rigidity. In this study, we show that efficient force transmission along SFs relies on actin crosslinking by alpha-actinin. Upon reduction of alpha-actinin-mediated crosslinks, the myosin II activity induced flows of actin filaments and myosin II along SFs, leading to a decrease in traction force exertion to ECM. The fluidized SFs maintained their cable integrity probably through enhanced actin polymerization throughout SFs. A computational modeling analysis suggested that lowering the density of actin crosslinks caused viscous slippage of actin filaments in SFs and, thereby, dissipated myosin-generated force transmitting along SFs. As a cellular scale outcome, alpha-actinin depletion attenuated the ECM-rigidity-dependent difference in cell migration speed, which suggested that alpha-actinin-modulated SF mechanics is involved in the cellular response to ECM rigidity.
Contact
Hiroaki Hirata
, Kanazawwa Institute of Technology
Contributors
Hiroki Katsuta, Satoru Okuda, Kazuaki Nagayama, Hiroaki Machiyama, Hiroaki Hirata
OMERO Project
Source