title: Chronic morphine treatment inhibits opioid receptor desensitization and internalization.
abstract: Chronic opioid receptor (OR) activation by morphine causes distinct cellular adaptations responsible for the development of tolerance. The present study examines the effect of chronic morphine exposure on the ability of high-efficacy agonists to mediate delta-OR (DOR) and mu-OR (MOR) uncoupling and internalization, two regulatory mechanisms contributing to rapid desensitization of OR function. Chronic morphine treatment (1 microm; 72 hr) of DOR carrying neuroblastoma x glioma (NG108-15) hybrid cells, a prototypical model system frequently used to study cellular aspects of opioid tolerance, completely blocked the capacity of [d-Ala2, d-Leu5]enkephalin (DADLE) and etorphine to desensitize opioid-stimulated [35S]GTPgammaS binding and to mediate DOR internalization. Similar findings were obtained on stably DOR- and MOR-transfected human embryonic kidney (HEK) 293 cells. Chronic morphine treatment also heterologously impaired agonist regulation of non-opioid G-protein-coupled receptors, such as the m(4)-muscarinic acetylcholine receptor and the brain-type cannabinoid receptor. As a possible underlying mechanism, we found that chronic morphine treatment completely blocked agonist-induced redistribution of beta-arrestin1 in both NG108-15 and stably MOR-transfected HEK293 cells. Moreover, attenuation of beta-arrestin1 function appears to depend on persistent stimulation of MAP kinase activity during the course of chronic morphine treatment, because coincubation of the cells together with the MAP kinase blocker PD98059 fully restored beta-arrestin1 translocation and receptor internalization. These results demonstrate that chronic morphine treatment produces adaptational changes at the beta-arrestin1 level, which in turn attenuates agonist-mediated desensitization and internalization of G-protein-coupled receptors.
authors.affiliation: Institute of Pharmacology, Toxicology and Pharmacy, University of Munich, D-80539 Munich, Germany. eisinger@pharmtox.vetmed.uni-muenchen.de
authors.forename: Daniela A
authors.initials: DA
authors.lastname: Eisinger
authors.forename: Hermann
authors.initials: H
authors.lastname: Ammer
authors.forename: Rüdiger
authors.initials: R
authors.lastname: Schulz
mesh_terms: D000818:Animals; D019390:Arrestins / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D016308:Enkephalin, Leucine-2-Alanine / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D019204:GTP-Binding Proteins / Q000378:metabolism; D006801:Humans; D006822:Hybrid Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D007476:Ionophores / Q000494:pharmacology; D007668:Kidney / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D020928:Mitogen-Activated Protein Kinases / Q000187:drug effects / Q000378:metabolism; D009020:Morphine / Q000494:pharmacology*; D009294:Narcotics / Q000494:pharmacology; D021381:Protein Transport / Q000187:drug effects; D043588:Receptor, Muscarinic M4; D043882:Receptors, Cannabinoid; D011956:Receptors, Cell Surface / Q000187:drug effects / Q000378:metabolism; D011955:Receptors, Drug / Q000235:genetics / Q000378:metabolism; D011976:Receptors, Muscarinic / Q000187:drug effects / Q000378:metabolism; D011957:Receptors, Opioid / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism*; D017465:Receptors, Opioid, delta / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D017450:Receptors, Opioid, mu / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D014162:Transfection; D000071557:beta-Arrestins
chemical_list: D019390:Arrestins; D004791:Enzyme Inhibitors; D007476:Ionophores; D009294:Narcotics; D043588:Receptor, Muscarinic M4; D043882:Receptors, Cannabinoid; D011956:Receptors, Cell Surface; D011955:Receptors, Drug; D011976:Receptors, Muscarinic; D011957:Receptors, Opioid; D017465:Receptors, Opioid, delta; D017450:Receptors, Opioid, mu; D000071557:beta-Arrestins; D016308:Enkephalin, Leucine-2-Alanine; D009020:Morphine; D020928:Mitogen-Activated Protein Kinases; D019204:GTP-Binding Proteins