Detail of fig1C_HeLa_AMPKAREV_2DG



Project
Title
FRET image of HeLa cells expressing AMPKAR-EV stimulated with 10mM 2-deoxyglucose
Description
NA
Release, Updated
2018-11-14
License
CC BY
Kind
Image data based on Experiment
File Formats
Data size
362.1 MB

Organism
H. sapiens ( NCBI:txid9606 )
Strain(s)
HeLa
Cell Line
-

Datatype
Cell dynamic
Molecular Function (MF)
Biological Process (BP)
AMP activated protein kinase activity ( GO:0004679 )
Cellular Component (CC)
nucleotide activated protein kinase complex ( GO:0031588 )
Biological Imaging Method
XYZ Scale
XY: 0.454 micrometer/pixel, Z: one slice
T scale
3 minutes for each time interval

Image Acquisition
Experiment type
TimeLapse
Microscope type
FluorescenceMicroscope
Acquisition mode
Other
Contrast method
Fluorescence
Microscope model
Olympus FV1000MVE or Olympus FV1200MPE-BX61WI
Detector model
two-channel GaAsP and two multialkali
Objective model
UplanSApo 30x/1.05 silicon or UplanSApo 25x/1.05 numerical aperture
Filter set

Summary of Methods
See details in Konagaya et al. (2017) Cell Rep, 21(9): 2628-2638.
Related paper(s)

Yumi Konagaya, Kenta Terai, Yusuke Hirao, Kanako Takakura, Masamichi Imajo, Yuji Kamioka, Norio Sasaoka, Akira Kakizuka, Kenta Sumiyama, Tomoichiro Asano, Michiyuki Matsuda (2017) A Highly Sensitive FRET Biosensor for AMPK Exhibits Heterogeneous AMPK Responses among Cells and Organs., Cell reports, Volume 21, Number 9, pp. 2628-2638

Published in 2017 Nov 28

(Abstract) AMP-activated protein kinase (AMPK), a master regulator of cellular metabolism, is a potential target for type 2 diabetes. Although extensive in vitro studies have revealed the complex regulation of AMPK, much remains unknown about the regulation in vivo. We therefore developed transgenic mice expressing a highly sensitive fluorescence resonance energy transfer (FRET)-based biosensor for AMPK, called AMPKAR-EV. AMPKAR-EV allowed us to readily examine the role of LKB1, a canonical stimulator of AMPK, in drug-induced activation and inactivation of AMPK in vitro. In transgenic mice expressing AMPKAR-EV, the AMP analog AICAR activated AMPK in muscle. In contrast, the antidiabetic drug metformin activated AMPK in liver, highlighting the organ-specific action of AMPK stimulators. Moreover, we found that AMPK was activated primarily in fast-twitch muscle fibers after tetanic contraction and exercise. These observations suggest that the AMPKAR-EV mouse will pave a way to understanding the heterogeneous responses of AMPK among cell types in vivo.
(MeSH Terms)

Contact
Kenta Terai , Kyoto University , Graduate School of Biostudies , Laboratory of Bioimaging and Cell Signaling
Contributors
Yumi Konagaya, Kenta Terai, Yusuke Hirao, Kanako Takakura, Masamichi Imajo, Yuji Kamioka, Norio Sasaoka, Akira Kakizuka, Kenta Sumiyama, Tomoichiro Asano, Michiyuki Matsuda

OMERO Dataset
OMERO Project
Source