Heterologous Regulation of Mu-Opioid (MOP) Receptor Mobility in the Membrane of SH-SY5Y Cells

Kévin Carayon,Lionel Moulédous,Anne Combedazou,Serge Mazères,Evert Haanappel,Laurence Salomé,Catherine Mollereau

doi:10.1074/jbc.m114.588558

Abstract

The dynamic organization of G protein-coupled receptors in the plasma membrane is suspected of playing a role in their function. The regulation of the diffusion mode of the mu-opioid (MOP) receptor was previously shown to be agonist-specific. Here we investigate the regulation of MOP receptor diffusion by heterologous activation of other G protein-coupled receptors and characterize the dynamic properties of the MOP receptor within the heterodimer MOP/neuropeptide FF (NPFF2) receptor. The data show that the dynamics and signaling of the MOP receptor in SH-SY5Y cells are modified by the activation of α2-adrenergic and NPFF2 receptors, but not by the activation of receptors not described to interact with the opioid receptor. By combining, for the first time, fluorescence recovery after photobleaching at variable radius experiments with bimolecular fluorescence complementation, we show that the MOP/NPFF2 heterodimer adopts a specific diffusion behavior that corresponds to a mix of the dynamic properties of both MOP and NPFF2 receptors. Altogether, the data suggest that heterologous regulation is accompanied by a specific organization of receptors in the membrane.

Highlights

MOP receptor function is presumably linked to a specific dynamic organization in the membrane
For the first time, fluorescence recovery after photobleaching at variable radius experiments with bimolecular fluorescence complementation, we show that the MOP/NPFF2 heterodimer adopts a specific diffusion behavior that corresponds to a mix of the dynamic properties of both MOP and NPFF2 receptors
The present study aimed to explore whether the functional heterologous regulation of G proteincoupled receptor (GPCR), in the case of heteromeric partners, is related to a specific dynamic behavior of receptors in the membrane, by considering the MOP receptor as an example

Summary

Introduction

MOP receptor function is presumably linked to a specific dynamic organization in the membrane. Results: Inhibition of MOP receptor signaling by NPFF2 and ␣2 receptors is accompanied by diffusion changes, with a particular behavior for heterodimers. Conclusion: MOP receptor function, diffusion, and confinement are subject to specific heterologous regulation by other GPCRs. Significance: Specific GPCR regulation is associated with particular dynamic organization in the membrane. We investigate the regulation of MOP receptor diffusion by heterologous activation of other G protein-coupled receptors and characterize the dynamic properties of the MOP receptor within the heterodimer MOP/neuropeptide FF (NPFF2) receptor. The data show that the dynamics and signaling of the MOP receptor in SH-SY5Y cells are modified by the activation of ␣2-adrenergic and NPFF2 receptors, but not by the activation of receptors not described to interact with the opioid receptor. The data suggest that heterologous regulation is accompanied by a specific organization of receptors in the membrane

Objectives

Results

Conclusion