SSBD:database

Key Organism Strain Cell Line Gene Symbol Protein name Protein tag Reporter Project ID Dataset ID Title Description Data Type Gene Ontology Biological Imaging Methods Method Summary Paper Information Medical Subject Headlines Person

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389-Watabe-PGE2discharge

https://ssbd.riken.jp/database/389-Watabe-PGE2discharge/

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ssbd-repos-000389

https://ssbd.riken.jp/repository/389/

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Visual quantification of prostaglandin E2 discharge from a single cell

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.

2026-01-21

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CC BY 4.0

Image data based on Experiment

146 ( Image datasets: 146, Quantitative data datasets: 0 )

77.0 GB ( Image datasets: 77.0 GB, Quantitative data datasets: 0 bytes )

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Homo sapiens, Canis lupus familiaris

HeLa cell, MDCK cell

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signal transduction, cell communication

cytoplasm

time lapse microscopy, FRET, fluorescence microscopy

NA, 1 minute

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Tetsuya Watabe, Shinya Yamahira, Michiyuki Matsuda, Kenta Terai (2023) Visual quantification of prostaglandin E(2) discharge from a single cell., Cell structure and function

Published in 2023 Oct 7 (Electronic publication in Oct. 7, 2023, midnight )

• doi: 10.1247/csf.23047
• pmid: 37813623
  

(Abstract) Calcium transients drive cells to discharge prostaglandin E(2) (PGE(2)). We visualized PGE(2)-induced protein kinase A (PKA) activation and quantitated PGE(2) secreted from a single cell by combining fluorescence microscopy and a simulation model. For this purpose, we first prepared PGE(2)-producer cells that express either an optogenetic or a chemogenetic calcium channel stimulator: OptoSTIM1 or Gq-DREADD, respectively. Second, we prepared reporter cells expressing the Gs-coupled PGE(2) reporter EP2 and the PKA biosensor Booster-PKA, which is based on the principle of Forster resonance energy transfer. Upon the stimulation-induced triggering of calcium transients, a single producer cell discharges PGE(2) 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 PGE(2) diffusion was restricted to the basolateral space. By fitting the radius of the PKA-activated area to a simulation model based on simple diffusion, we estimated that a single HeLa cell secretes 0.25 fmol PGE(2) upon a single calcium transient to activate PKA in more than 1000 neighboring cells. This model also predicts that the PGE(2) discharge rate is comparable to the diffusion rate. Thus, our method quantitatively envisions that a single calcium transient affects more than 1000 neighboring cells via PGE(2).Keywords: prostaglandin E(2), imaging, intercellular communication, biosensor, quantification.

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

Tetsuya Watabe, Shinya Yamahira

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