Detail of Figure3G_invitro_N1_DP



Project
Title
Fluorescence images of the ERK activity of isolated DP (CD4/CD8 double-positive) thymocytes in vitro
Description
Fluorescence images of the ERK activity of isolated DP (CD4/CD8 double-positive) thymocytes in vitro
Release, Updated
2021-09-30
License
CC BY
Kind
Image data
File Formats
Data size
2.0 MB

Organism
Mus musculus ( NCBI:txid10090 )
Strain(s)
-
Cell Line
-
Protein names
ERK

Datatype
-
Molecular Function (MF)
-
Biological Process (BP)
activation of MAPERK kinase ( GO:0000186 ) cell motility ( GO:0048870 )
Cellular Component (CC)
T cell receptor complex ( GO:0042101 )
Biological Imaging Method
fluorescence microscopy ( Fbbi:00000246 )
X scale
0.795 micrometer/pixel
Y scale
0.795 micrometer/pixel
Z scale
1 micrometer/slice
T scale
-

Image Acquisition
Experiment type
-
Microscope type
-
Acquisition mode
-
Contrast method
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Microscope model
-
Detector model
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Objective model
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Filter set
-

Summary of Methods
See details in Konishi Y, et. al. (2018) iScience, 10: 98-113.
Related paper(s)

Yoshinobu Konishi, Kenta Terai, Yasuhide Furuta, Hiroshi Kiyonari, Takaya Abe, Yoshihiro Ueda, Tatsuo Kinashi, Yoko Hamazaki, Akifumi Takaori-Kondo, Michiyuki Matsuda (2018) Live-Cell FRET Imaging Reveals a Role of Extracellular Signal-Regulated Kinase Activity Dynamics in Thymocyte Motility., iScience, Volume 10, pp. 98-113

Published in 2018 Dec 21 (Electronic publication in Nov. 20, 2018, midnight )

(Abstract) Extracellular signal-regulated kinase (ERK) plays critical roles in T cell development in the thymus. Nevertheless, the dynamics of ERK activity and the role of ERK in regulating thymocyte motility remain largely unknown due to technical limitations. To visualize ERK activity in thymocytes, we here developed knockin reporter mice expressing a Forster/fluorescence resonance energy transfer (FRET)-based biosensor for ERK from the ROSA26 locus. Live imaging of thymocytes isolated from the reporter mice revealed that ERK regulates thymocyte motility in a subtype-specific manner. Negative correlation between ERK activity and motility was observed in CD4/CD8 double-positive thymocytes and CD8 single-positive thymocytes, but not in CD4 single-positive thymocytes. Interestingly, however, the temporal deviations of ERK activity from the average correlate with the motility of CD4 single-positive thymocytes. Thus, live-cell FRET imaging will open a window to understanding the dynamic nature and the diverse functions of ERK signaling in T cell biology.

Contact
Kenta Terai , Kyoto University , Graduate School of Biostudies , Laboratory of Bioimaging and Cell Signaling
Contributors

OMERO Dataset
OMERO Project
Source