Visual quantification of prostaglandin E₂ discharge from a single cell.

ABSTRACT: Calcium transients drive cells to discharge prostaglandin E₂ (PGE₂). We 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, we first prepared PGE₂-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 PGE₂ reporter EP2 and the PKA biosensor Booster-PKA, which is based on the principle of Förster resonance energy transfer (FRET). Upon the stimulation-induced triggering of calcium transients, a single producer cell discharges PGE₂ to stimulate PK

SUBMITTER: Watabe T 

PROVIDER: S-EPMC11496778 | biostudies-literature | 2023 Dec

REPOSITORIES: biostudies-literature