Pharmacological and phosphoproteomic approaches to roles of protein kinase C in kappa opioid receptor-mediated effects in mice

Neuropharmacology. 2020 Dec 15:181:108324. doi: 10.1016/j.neuropharm.2020.108324. Epub 2020 Sep 22.

Abstract

Kappa opioid receptor (KOR) agonists possess adverse dysphoric and psychotomimetic effects, thus limiting their applications as non-addictive anti-pruritic and analgesic agents. Here, we showed that protein kinase C (PKC) inhibition preserved the beneficial antinociceptive and antipruritic effects of KOR agonists, but attenuated the adverse condition placed aversion (CPA), sedation, and motor incoordination in mice. Using a large-scale mass spectrometry-based phosphoproteomics of KOR-mediated signaling in the mouse brain, we observed PKC-dependent modulation of G protein-coupled receptor kinases and Wnt pathways at 5 min; stress signaling, cytoskeleton, mTOR signaling and receptor phosphorylation, including cannabinoid receptor CB1 at 30 min. We further demonstrated that inhibition of CB1 attenuated KOR-mediated CPA. Our results demonstrated the feasibility of in vivo biochemical dissection of signaling pathways that lead to side effects.

Keywords: CB1; Phosphoproteomics; Protein kinase C; Wnt; mTOR; κ opioid receptor.

Publication types

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

MeSH terms

  • 3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cyclohexyl)-benzeneacetamide, (trans)-Isomer / pharmacology
  • Animals
  • Avoidance Learning
  • G-Protein-Coupled Receptor Kinases
  • Male
  • Mice
  • Motor Activity / drug effects
  • Phosphoproteins
  • Phosphorylation
  • Protein Kinase C / drug effects
  • Protein Kinase C / genetics*
  • Protein Kinase C / radiation effects
  • Protein Kinase Inhibitors
  • Proteomics
  • Receptor, Cannabinoid, CB1 / drug effects
  • Receptors, G-Protein-Coupled / drug effects
  • Receptors, Opioid, kappa / drug effects
  • Receptors, Opioid, kappa / genetics*
  • Receptors, Opioid, kappa / radiation effects
  • Signal Transduction / drug effects*
  • TOR Serine-Threonine Kinases / drug effects
  • Wnt Signaling Pathway / drug effects

Substances

  • Phosphoproteins
  • Protein Kinase Inhibitors
  • Receptor, Cannabinoid, CB1
  • Receptors, G-Protein-Coupled
  • Receptors, Opioid, kappa
  • 3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cyclohexyl)-benzeneacetamide, (trans)-Isomer
  • mTOR protein, mouse
  • TOR Serine-Threonine Kinases
  • Protein Kinase C
  • G-Protein-Coupled Receptor Kinases