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Cell Rep. 2017 Nov 28;21(9):2628-2638. doi: 10.1016/j.celrep.2017.10.113.

A Highly Sensitive FRET Biosensor for AMPK Exhibits Heterogeneous AMPK Responses among Cells and Organs.

Author information

1
Laboratory of Bioimaging and Cell Signaling, Graduate School of Biostudies, Kyoto University, Kyoto 606-8501, Japan.
2
Laboratory of Bioimaging and Cell Signaling, Graduate School of Biostudies, Kyoto University, Kyoto 606-8501, Japan. Electronic address: terai.kenta.5m@kyoto-u.ac.jp.
3
Department of Medical Science, Graduate School of Medicine, University of Hiroshima, Hiroshima 734-8553, Japan.
4
Imaging Platform for Spatio-Temporal Regulation, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan.
5
Department of Molecular Genetics, Institute of Biomedical Science, Kansai Medical University, Osaka 570-8506, Japan.
6
Laboratory of Functional Biology, Graduate School of Biostudies, Kyoto University, Kyoto 606-8501, Japan.
7
Laboratory for Mouse Genetic Engineering, Quantitative Biology Center, RIKEN, Osaka 565-0874, Japan.
8
Laboratory of Bioimaging and Cell Signaling, Graduate School of Biostudies, Kyoto University, Kyoto 606-8501, Japan; Department of Pathology and Biology of Diseases, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan.

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.

KEYWORDS:

AMP-activated protein kinase; AMPK; FRET; LKB1; Pin1; cell metabolism; fluorescence resonance energy transfer; live imaging; liver kinase B1; peptidyl-prolyl cis/trans isomerase NIMA-interacting 1

PMID:
29186696
DOI:
10.1016/j.celrep.2017.10.113
[Indexed for MEDLINE]
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