Endocytosis and Compartmentalized Intracellular Signaling of the Prostaglandin Receptor EP4 Mediate Pain
- 1New York University
Abstract
Prostaglandin E2 (PGE2) is recognized as a major mediator of inflammatory pain. However, the intracellular signaling mechanisms by which PGE2 mediates pain remain unclear. Here, we show that pain-like responses evoked by PGE2 in dorsal root ganglion (DRG) nociceptors are mediated by internalization and compartmentalized signaling of the EP4 receptor. The EP4-selective agonist L-902,688 induced immediate nocifensive behavior and prolonged mechanical allodynia in mice and sensitized isolated DRG nociceptors. Pharmacological inhibition of clathrin- and dynamin-mediated endocytosis (using pitstop2 and dyngo4a) and siRNA knockdown of Dnm1 prevented EP4-induced nociception and sensitization, implicating receptor trafficking in pain signaling. Using genetically-encoded biosensors, we monitored EP4 trafficking and downstream signaling within subcellular compartments of HEK293 cells. PGE2 stimulated dynamin- and {beta}-arrestin-dependent EP4 trafficking from the plasma membrane to early, late and recycling endosomes and Golgi apparatus, and mobilized intracellular EP4 pools from the endoplasmic reticulum. PGE2 induced the assembly of EP4, G proteins and {beta}-arrestin signaling complexes in endosomes and the Golgi apparatus. Inhibition of EP4 endocytosis suppressed intracellular cAMP production and ERK activation. Our findings reveal that EP4-mediated nociceptive signaling originates from intracellular compartments and suggest that modulating EP4 internalization and subcellular signaling dynamics could offer a novel strategy for inflammatory pain management.
Related
- A stochastic model of compartmentalized intracellular signalling
- Anti-HER2 immunotoxin scFv-PE35KDEL internalizes via receptor-mediated endocytosis and induces apoptosis in ovarian cancer cells
- Metabotropic glutamate receptor internalization and synaptic AMPA receptor endocytosis by dopamine
- From First Bond to Stable Adhesion: Multivalent Bond Stabilization in Receptor-Mediated Endocytosis
- Endocytosis and Endocytic Neurotrophin Signaling in Developing Spiral Ganglion Neurons.
BibTeX
@misc{sokrat2026endocytosis,
title = {{Endocytosis and Compartmentalized Intracellular Signaling of the Prostaglandin Receptor EP4 Mediate Pain}},
author = {B. Sokrat and R. Tonello and P. Duran and N. Barrett and Y. Mulpuri and B. L. Schmidt and D. D. Jensen and F. De Logu and R. Nassini and P. Geppetti and N. W. Bunnett},
howpublished = {bioRxiv},
year = {2026},
month = sep,
doi = {10.64898/2026.09.18.752773},
url = {https://www.biorxiv.org/content/10.64898/2026.09.18.752773v1},
note = {CC BY},
}
RIS
TY - UNPB
TI - Endocytosis and Compartmentalized Intracellular Signaling of the Prostaglandin Receptor EP4 Mediate Pain
AU - B. Sokrat
AU - R. Tonello
AU - P. Duran
AU - N. Barrett
AU - Y. Mulpuri
AU - B. L. Schmidt
AU - D. D. Jensen
AU - F. De Logu
AU - R. Nassini
AU - P. Geppetti
AU - N. W. Bunnett
PY - 2026
DA - 2026/09/18
PB - bioRxiv
DO - 10.64898/2026.09.18.752773
UR - https://www.biorxiv.org/content/10.64898/2026.09.18.752773v1
AB - Prostaglandin E2 (PGE2) is recognized as a major mediator of inflammatory pain. However, the intracellular signaling mechanisms by which PGE2 mediates pain remain unclear. Here, we show that pain-like responses evoked by PGE2 in dorsal root ganglion (DRG) nociceptors are mediated by internalization and compartmentalized signaling of the EP4 receptor. The EP4-selective agonist L-902,688 induced immediate nocifensive behavior and prolonged mechanical allodynia in mice and sensitized isolated DRG nociceptors. Pharmacological inhibition of clathrin- and dynamin-mediated endocytosis (using pitstop2 and dyngo4a) and siRNA knockdown of Dnm1 prevented EP4-induced nociception and sensitization, implicating receptor trafficking in pain signaling. Using genetically-encoded biosensors, we monitored EP4 trafficking and downstream signaling within subcellular compartments of HEK293 cells. PGE2 stimulated dynamin- and {beta}-arrestin-dependent EP4 trafficking from the plasma membrane to early, late and recycling endosomes and Golgi apparatus, and mobilized intracellular EP4 pools from the endoplasmic reticulum. PGE2 induced the assembly of EP4, G proteins and {beta}-arrestin signaling complexes in endosomes and the Golgi apparatus. Inhibition of EP4 endocytosis suppressed intracellular cAMP production and ERK activation. Our findings reveal that EP4-mediated nociceptive signaling originates from intracellular compartments and suggest that modulating EP4 internalization and subcellular signaling dynamics could offer a novel strategy for inflammatory pain management.
ER -