Detail of Figure4A_2pabPAC_20200127_1st_z41_2P



Project
Title
FRET images of the hepatocytes in the livers of Booster-PKAchu mice with 2pabPAC expression and activated by 2P excitation.
Description
FRET images of the hepatocytes in the livers of Booster-PKAchu mice with 2pabPAC expression and activated by 2P excitation.
Release, Updated
2022-03-31
License
CC-BY
Kind
Image data
File Formats
.oib
Data size
44.2 MB

Organism
Mus musculus ( NCBITaxon:10090 )
Strain(s)
-
Cell Line
-

Datatype
-
Molecular Function (MF)
-
Biological Process (BP)
cAMP-mediated signaling ( GO:0019933 )
Cellular Component (CC)
cAMP-dependent protein kinase complex ( GO:0005952 )
Biological Imaging Method
X scale
0.795 micrometer/pixel
Y scale
0.795 micrometer/pixel
Z scale
2 micrometer/pixel
T scale
30 sec per time interval

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

Summary of Methods
See details in Kinjo T, et. al. (2020) ACS Chem Biol., 15(11):2848-2853.
Related paper(s)

Tomoaki Kinjo, Tetsuya Watabe, Kenju Kobachi, Kenta Terai, Michiyuki Matsuda (2020) Single-Cell Activation of the cAMP-Signaling Pathway in 3D Tissues with FRET-Assisted Two-Photon Activation of bPAC., ACS chemical biology, Volume 15, Number 11, pp. 2848-2853

Published in 2020 Nov 20 (Electronic publication in Oct. 19, 2020, midnight )

(Abstract) Bacterial photoactivated adenylyl cyclase (bPAC) has been widely used in signal transduction research. However, due to its low two-photon absorption, bPAC cannot be efficiently activated by two-photon (2P) excitation. Taking advantage of the high two-photon absorption of monomeric teal fluorescent protein 1 (mTFP1), we herein developed 2P-activatable bPAC (2pabPAC), a fusion protein consisting of bPAC and mTFP1. In 2pabPAC, the energy absorbed by mTFP1 excites bPAC by Furster resonance energy transfer (FRET) at ca. 43% efficiency. The light-induced increase in cAMP was monitored by a red-shifted FRET biosensor for PKA. In 3D MDCK cells and mouse liver, PKA was activated at single-cell resolution under a 2P microscope. We found that PKA activation in a single hepatocyte caused PKA activation in neighboring cells, indicating the propagation of PKA activation. Thus, 2pabPAC will provide a versatile platform for controlling the cAMP signaling pathway and investigating cell-to-cell communication in vivo.
(MeSH Terms)

Contact
Kenta Terai , Graduate School of Medicine, Kyoto University , Department of Pathology and Biology of Diseases
Contributors

OMERO Dataset
OMERO Project
Source