Laura Glück1, Anastasia Loktev1, Lionel Moulédous2, Catherine Mollereau2, Ping-Yee Law3, Stefan Schulz4. 1. Institute of Pharmacology and Toxicology, Jena University Hospital, Friedrich Schiller University Jena, Jena, Germany. 2. Institut de Pharmacologie et de Biologie Structurale, CNRS/Université de Toulouse, Toulouse Cedex, France. 3. Department of Pharmacology, University of Minnesota Medical School, Minneapolis, Minnesota. 4. Institute of Pharmacology and Toxicology, Jena University Hospital, Friedrich Schiller University Jena, Jena, Germany. Electronic address: Stefan.Schulz@med.uni-jena.de.
Abstract
BACKGROUND: The clinical benefits of opioid drugs are counteracted by the development of tolerance and addiction. We provide in vivo evidence for the involvement of G protein-coupled receptor kinases (GRKs) in opioid dependence in addition to their roles in agonist-selective mu-opioid receptor (MOR) phosphorylation. METHODS: In vivo MOR phosphorylation was examined by immunoprecipitation and nanoflow liquid chromatography-tandem mass spectrometry analysis. Using the hot-plate and conditioned place preference test, we investigated opioid-related antinociception and reward effects in mice lacking GRK3 or GRK5. RESULTS: Etonitazene and fentanyl stimulated the in vivo phosphorylation of multiple carboxyl-terminal phosphate acceptor sites, including threonine 370, serine 375, and threonine 379, which was predominantly mediated by GRK3. By contrast, morphine promoted a selective phosphorylation of serine 375 that was predominantly mediated by GRK5. In contrast to GRK3 knockout mice, GRK5 knockout mice exhibited reduced antinociceptive responses after morphine administration and developed morphine tolerance similar to wild-type mice but fewer signs of physical dependence. Also, morphine was ineffective in inducing conditioned place preference in GRK5 knockout mice, whereas cocaine conditioned place preference was retained. However, the reward properties of morphine were evident in knock-in mice expressing a phosphorylation-deficient S375A mutation of the MOR. CONCLUSIONS: These findings show for the first time that MOR phosphorylation is regulated by agonist-selective recruitment of distinct GRK isoforms that influence different opioid-related behaviors. Modulation of GRK5 function could serve as a new approach for preventing addiction to opioids, while maintaining the analgesic properties of opioid drugs at an effective level.
BACKGROUND: The clinical benefits of opioid drugs are counteracted by the development of tolerance and addiction. We provide in vivo evidence for the involvement of G protein-coupled receptor kinases (GRKs) in opioid dependence in addition to their roles in agonist-selective mu-opioid receptor (MOR) phosphorylation. METHODS: In vivo MOR phosphorylation was examined by immunoprecipitation and nanoflow liquid chromatography-tandem mass spectrometry analysis. Using the hot-plate and conditioned place preference test, we investigated opioid-related antinociception and reward effects in mice lacking GRK3 or GRK5. RESULTS:Etonitazene and fentanyl stimulated the in vivo phosphorylation of multiple carboxyl-terminal phosphate acceptor sites, including threonine 370, serine 375, and threonine 379, which was predominantly mediated by GRK3. By contrast, morphine promoted a selective phosphorylation of serine 375 that was predominantly mediated by GRK5. In contrast to GRK3 knockout mice, GRK5 knockout mice exhibited reduced antinociceptive responses after morphine administration and developed morphine tolerance similar to wild-type mice but fewer signs of physical dependence. Also, morphine was ineffective in inducing conditioned place preference in GRK5 knockout mice, whereas cocaine conditioned place preference was retained. However, the reward properties of morphine were evident in knock-in mice expressing a phosphorylation-deficient S375A mutation of the MOR. CONCLUSIONS: These findings show for the first time that MOR phosphorylation is regulated by agonist-selective recruitment of distinct GRK isoforms that influence different opioid-related behaviors. Modulation of GRK5 function could serve as a new approach for preventing addiction to opioids, while maintaining the analgesic properties of opioid drugs at an effective level.
Authors: Sudha K Shenoy; Matthew T Drake; Christopher D Nelson; Daniel A Houtz; Kunhong Xiao; Srinivasan Madabushi; Eric Reiter; Richard T Premont; Olivier Lichtarge; Robert J Lefkowitz Journal: J Biol Chem Date: 2005-11-09 Impact factor: 5.157
Authors: Paul C Scherer; Nicholas W Zaccor; Neil M Neumann; Chirag Vasavda; Roxanne Barrow; Andrew J Ewald; Feng Rao; Charlotte J Sumner; Solomon H Snyder Journal: Proc Natl Acad Sci U S A Date: 2017-12-04 Impact factor: 11.205
Authors: Anika Mann; Lionel Moulédous; Carine Froment; Patrick R O'Neill; Pooja Dasgupta; Thomas Günther; Gloria Brunori; Brigitte L Kieffer; Lawrence Toll; Michael R Bruchas; Nurulain T Zaveri; Stefan Schulz Journal: Sci Signal Date: 2019-03-26 Impact factor: 8.192
Authors: Lawrence Toll; Michael R Bruchas; Girolamo Calo'; Brian M Cox; Nurulain T Zaveri Journal: Pharmacol Rev Date: 2016-03-08 Impact factor: 25.468
Authors: Renee A Bouley; Zara Y Weinberg; Helen V Waldschmidt; Yu-Chen Yen; Scott D Larsen; Manojkumar A Puthenveedu; John J G Tesmer Journal: Mol Pharmacol Date: 2020-03-31 Impact factor: 4.436