Organized cannabinoid receptor distribution in neurons revealed by super-resolution fluorescence imaging

G-protein-coupled receptors (GPCRs) play important roles in cellular functions. However, their intracellular organization is largely unknown. Through investigation of the cannabinoid receptor 1 (CB₁), we discovered periodically repeating clusters of CB₁ hotspots within the axons of neurons. We observed these CB₁ hotspots interact with the membrane-associated periodic skeleton (MPS) forming a complex crucial in the regulation of CB₁ signaling. Furthermore, we found that CB₁ hotspot periodicity increased upon CB₁ agonist application, and these activated CB₁ displayed less dynamic movement compared to non-activated CB₁. Our results suggest that CB₁ forms periodic hotspots organized by the MPS as a mechanism to increase signaling efficacy upon activation.