389-Watabe-PGE2discharge
Author reviewedVisual quantification of prostaglandin E2 discharge from a single cell
Project ID
389-Watabe-PGE2discharge
Project SID
389
Title
Visual quantification of prostaglandin E2 discharge from a single cell
Description
Calcium transients drive cells to discharge prostaglandin E2 (PGE2). The authors visualized PGE2-induced protein kinase A (PKA) activation and quantitated PGE2 secreted from a single cell by combining fluorescence microscopy and a simulation model. For this purpose, they first prepared PGE2-producer cells that express either an optogenetic or a chemogenetic calcium channel stimulator: OptoSTIM1 (CRY2clust) or Gq-DREADD, respectively. Second, they prepared reporter cells expressing the Gs-coupled PGE2 reporter EP2 and the PKA biosensor Booster-PKA, which is based on the principle of Forster resonance FRET. Upon the stimulation-induced triggering of calcium transients, a single producer cell discharges PGE2 to stimulate PKA in the surrounding reporter cells. Due to the flow of the medium, the PKA-activated area exhibited a comet-like smear when HeLa cells were used. In contrast, radial PKA activation was observed when confluent MDCK cells were used, indicating that PGE2 diffusion was restricted to the basolateral space. By fitting the radius of the PKA-activated area to a simulation model based on simple diffusion, the authors estimated that a single HeLa cell secretes 0.25 fmol PGE2 upon a single calcium transient to activate PKA in more than 1000 neighboring cells. This model also predicts that the PGE2 discharge rate is comparable to the diffusion rate. Thus, their method quantitatively envisions that a single calcium transient affects more than 1000 neighboring cells via PGE2.
Submission Date
2024-08-30
Opened Date
-
Release Date
2026-01-21
Update Date
-
Kind
Image datasets
License
Project URL
-
Project DOI
-
Metadata Version
-
Template Version
-
Funding Information
Financial
support was provided by JSPS KAKENHI grants (nos. 21K20773 to
T.W., 21H02715 and 21H05226 to K.T., and 19H00993, 20H05898,
22H04926 to M.M.), a JST Moonshot R&D grant (no. JPMJPS2022
to M.M.), a grant from Fugaku Foundation to M.M., and Research
Grants of Princess Takamatsu Cancer Research Fund to K.T.
Total
12063 files / 287.6 GB
Image datasets
zipped 146 files / 62.5 GB, raw image 689 files / 77.0 GB, total 835 files / 139.5 GB
Quantitative datasets
zipped 0 files / 0 bytes, raw bdml 0 files / 0 bytes, total 0 files / 0 bytes
Biosample
Organism
Homo sapiens
(
NCBITaxon:9606
)
Canis lupus familiaris
(
NCBITaxon:9615
)
Strain
-
Cell
-
Cell line
HeLa cell
(
CLO:0003684
)
MDCK cell
(
CLO:0007646
)
MeSH
HeLa Cells
(
D008856,
D059008,
D031541
)
Cells
Cell Communication,
Signal Transduction
Cytoplasm
Madin Darby Canine Kidney Cells
(
D008856,
D059008,
D031541
)
HeLa Cells
(
D008856,
D031541
)
Ontology
Anatomical Entity
-
UBERON
cell
(
CL:0000000
)
Biological Process
cell communication,
signal transduction
(
GO:0007165
)
Cellular Component
cytoplasm
(
GO:0005737
)
Molecular Function
-
Medical Subject Headings
HeLa Cells
(
D008856,
D059008,
D031541
)
Cells
Cell Communication,
Signal Transduction
Cytoplasm
Madin Darby Canine Kidney Cells
(
D008856,
D059008,
D031541
)
HeLa Cells
(
D008856,
D031541
)
Imaging Method
Method involved in biological imaging
fluorescence microscopy
(
FBbi:00000246
)
time lapse microscopy
(
FBbi:00000249
)
FRET
(
FBbi:00000367
)
Paper DOI / Paper URL
People
Contact
Kenta Terai
Kyoto University
Laboratory of Bioimaging and Cell Signaling, Graduate School of Biostudies
-
-
Imaging dataset contributor
Tetsuya Watabe (Kyoto University)
Shinya Yamahira (Kyoto University)
Kenta Terai (Kyoto University)
Quantitative dataset contributor
-
Image datasets
146
Quantitative datasets
0
Dataset List
Thumbnail
Dataset
Organism
Kind / Links
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-HeLa cells cocultured with producer HeLa cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-HeLa cells cocultured with producer HeLa cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
FRET images of Booster-PKA in reporter COX-DKO-HeLa cells treated with 0.1 nM PGE2
FRET images of Booster-PKA in reporter COX-DKO-HeLa cells treated with 0.3 nM PGE2
FRET images of Booster-PKA in reporter COX-DKO-HeLa cells treated with 0 nM PGE2
FRET images of Booster-PKA in reporter COX-DKO-HeLa cells treated with 100 nM PGE2
FRET images of Booster-PKA in reporter COX-DKO-HeLa cells treated with 10 nM PGE2
FRET images of Booster-PKA in reporter COX-DKO-HeLa cells treated with 1 nM PGE2
FRET images of Booster-PKA in reporter COX-DKO-HeLa cells treated with 3 nM PGE2
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-HeLa cells cocultured with producer HeLa cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-HeLa cells cocultured with producer HeLa cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-HeLa cells cocultured with producer HeLa cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-HeLa cells cocultured with producer HeLa cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-HeLa cells cocultured with producer HeLa cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-HeLa cells cocultured with producer HeLa cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-HeLa cells cocultured with producer HeLa cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-HeLa cells cocultured with producer HeLa cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells treated with EGF
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells treated with EGF
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells treated with EGF
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells treated with EGF
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells treated with EGF
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells treated with EGF
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells treated with EGF
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells treated with EGF
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells treated with EGF
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells treated with EGF
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells treated with EGF
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells treated with EGF
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells treated with EGF
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells treated with EGF
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells treated with MEK inhibitor
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells treated with MEK inhibitor
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells treated with MEK inhibitor
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells treated with MEK inhibitor
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells treated with MEK inhibitor
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells treated with MEK inhibitor
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells treated with MEK inhibitor
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells treated with MEK inhibitor
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells treated with MEK inhibitor
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells treated with MEK inhibitor
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells treated with MEK inhibitor
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells treated with MEK inhibitor
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells treated with MEK inhibitor
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells treated with MEK inhibitor
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells
Time-lapse FRET images of reporter HeLa cells and producer HeLa cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-HeLa cells expressing EP2 cocultured with producer HeLa cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-HeLa cells expressing EP2 cocultured with producer HeLa cells in the presence of EP2/EP4 inhibitor
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells cocultured with producer MDCK cells in the presence of EP2/EP4 inhibitor
Time-lapse FRET images of Booster-PKA in reporter COX-DKO-MDCK cells that were deficient in E-cadherin, and were cocultured with producer MDCK cells