A Genetically Encoded Biosensor Reveals Location Bias of Opioid Drug Action

Neuron. 2018 Jun 6;98(5):963-976.e5. doi: 10.1016/j.neuron.2018.04.021. Epub 2018 May 10.

Abstract

Opioid receptors (ORs) precisely modulate behavior when activated by native peptide ligands but distort behaviors to produce pathology when activated by non-peptide drugs. A fundamental question is how drugs differ from peptides in their actions on target neurons. Here, we show that drugs differ in the subcellular location at which they activate ORs. We develop a genetically encoded biosensor that directly detects ligand-induced activation of ORs and uncover a real-time map of the spatiotemporal organization of OR activation in living neurons. Peptide agonists produce a characteristic activation pattern initiated in the plasma membrane and propagating to endosomes after receptor internalization. Drugs produce a different activation pattern by additionally driving OR activation in the somatic Golgi apparatus and Golgi elements extending throughout the dendritic arbor. These results establish an approach to probe the cellular basis of neuromodulation and reveal that drugs distort the spatiotemporal landscape of neuronal OR activation.

Keywords: GPCR; Golgi; biosensor; endosome; ligand bias; ligand-induced activation; opioid drug; opioid receptor; signaling; subcellular location.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Video-Audio Media

MeSH terms

  • Analgesics, Opioid / metabolism*
  • Animals
  • Biosensing Techniques
  • Cell Membrane / metabolism*
  • Dendrites / metabolism*
  • Endosomes / metabolism*
  • Enkephalin, Ala(2)-MePhe(4)-Gly(5)- / metabolism
  • Enkephalin, D-Penicillamine (2,5)- / metabolism
  • Enkephalin, Leucine-2-Alanine / metabolism
  • Golgi Apparatus / metabolism*
  • HEK293 Cells
  • HeLa Cells
  • Humans
  • Intracellular Space
  • Microscopy, Fluorescence
  • Morphine / metabolism
  • Naloxone
  • Narcotic Antagonists
  • Neurons / metabolism*
  • Peptides / metabolism*
  • Rats
  • Receptors, Opioid / metabolism*
  • Spatio-Temporal Analysis

Substances

  • Analgesics, Opioid
  • Narcotic Antagonists
  • Peptides
  • Receptors, Opioid
  • Enkephalin, Ala(2)-MePhe(4)-Gly(5)-
  • Naloxone
  • Enkephalin, Leucine-2-Alanine
  • Morphine
  • Enkephalin, D-Penicillamine (2,5)-