Dataset Fig5c_TensionCaluculinA
Project
380-Wang-ActininTensionSensor
Title
Changes of tension in smooth muscle cell after treatment with water, calyculin A, or Y27632
Description
FRET-based sensors are utilized for real-time measurements of cellular tension. However, transfection of the sensor gene shows low efficacy and is only effective for a short period. Reporter mice expressing such sensors have been developed, but sensor fluorescence has not been measured successfully using conventional confocal microscopy. Therefore, methods for spatiotemporal measurement of cellular tension in vivo or ex vivo are still limited. The authors established a reporter mouse line expressing FRET-based actinin tension sensors consisting of EGFP as the donor and mCherry as the acceptor and whose FRET ratio change is observable with confocal microscopy. Tension-induced changes in FRET signals were monitored in the aorta and tail tendon fascicles, as well as aortic smooth muscle cells isolated from these mice. The pattern of FRET changes was distinctive, depending on tissue type. Indeed, aortic smooth muscle cells exhibit different sensitivity to macroscopic tensile strain in situ and in an isolated state. This mouse strain will enable novel types of biomechanical investigations of cell functions in important physiological events.
Funding
AMED-CREST from the Japan Agency for Medical Research and Development (JP19gm0810005), the NAKATANI Foundation, and JSPS KAKENHIs (Nos. 21H04533 and 21K19902)
Title
Time-lapse images of tension sensor in VSMCs isolated from the aorta before and after stimulation with calyculin A
Description
Time-lapse images of actinin tension sensor in VSMCs (Vascular smooth muscle cells) isolated from the aorta before and after stimulation with calyculin A. Decrease in energy transfer from EGFP to mCherry reflects the increase in cell tension. Calyculin A was added between 21-22 min.
Description
Vascular smooth muscle cells isolated from the aorta in R26-S2 mice expressing actinin tension sensor
Cells
vascular associated smooth muscle cell (
CL_0000359)
Intrinsic variables
The reporter mouse line, R26R-S2 expressing actinin tension sensor with which monitors a degree of cellular tension by FRET imaging
Extrinsic variables
calyculin A
Gene ontology: Biological processes
Gene ontology: Cellular components
Gene ontology: Molecular functions
Dimensions
512x512x10x2x63
X scales
0.264 micrometer/pixel
Y scales
0.264 micrometer/pixel
Z scales
0.5 micrometer/slice
T scales
1 minutes per time interval
Microscopy types
FRET (FBbi:00000367), confocal microscopy (Fbbi:00000251), time lapse microscopy (
FBbi:00000249)
Detection methods
wide-field detection
Visualization methods
EGFP, mCherry
Illumination methods
wide-field detection
Contrast enhancing methods
Resolution enhancing methods
Sample preparation methods
living tissue
Light source
Argon laser 488 nm
Objective
Plan-Apochromat 63x/1.4 Oil DIC M27