Morphine-induced mu-opioid receptor rapid desensitization is independent of receptor phosphorylation and beta-arrestins.
Receptor desensitization involving receptor phosphorylation and subsequent betaArrestin (betaArr) recruitment has been implicated in the tolerance development mediated by mu-opioid receptor (OPRM1). However, the roles of receptor phosphorylation and betaArr on morphine-induced OPRM1 desensitization remain to be demonstrated. Using OPRM1-induced intracellular Ca(2+) ([Ca(2+)](i))release to monitor receptor activation, as predicted, [D-Ala(2), N-Me-Phe(4), Gly(5)-ol]-enkephalin (DAMGO), induced OPRM1 desensitization in a receptor phosphorylation- and betaArr-dependent manner. The DAMGO-induced OPRM1 desensitization was attenuated significantly when phosphorylation deficient OPRM1 mutants or Mouse Embryonic Fibroblast (MEF) cells from betaArr1 and 2 knockout mice were used in the studies. Specifically, DAMGO-induced desensitization was blunted in HEK293 cells expressing the OPRM1S375A mutant and was eliminated in MEF cells isolated from betaArr2 knockout mice expressing the wild type OPRM1. However, although morphine also could induce a rapid desensitization on [Ca(2+)](i) release to a greater extent than that of DAMGO and could induce the phosphorylation of Ser(375) residue, morphine-induced desensitization was not influenced by mutating the phosphorylation sites or in MEF cells lacking betaArr1 and 2. Hence, morphine could induce OPRM1 desensitization via pathway independent of betaArr, thus suggesting the in vivo tolerance development to morphine can occur in the absence of betaArr.
Department of Pharmacology, Medical School, University of Minnesota, Minneapolis, MN 55455-0217, USA. chux0086@umn.edu
Ji
J
Chu
Hui
H
Zheng
Horace H
HH
Loh
Ping-Yee
PY
Law
D000818:Animals; D019390:Arrestins / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D020875:Enkephalin, Ala(2)-MePhe(4)-Gly(5)- / Q000494:pharmacology; D006801:Humans; D042541:Intracellular Space / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D009020:Morphine / Q000494:pharmacology*; D050505:Mutant Proteins / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D017450:Receptors, Opioid, mu / Q000819:agonists / Q000378:metabolism*; D018048:Receptors, Purinergic P2 / Q000378:metabolism; D000071557:beta-Arrestins
D019390:Arrestins; D050505:Mutant Proteins; D017450:Receptors, Opioid, mu; D018048:Receptors, Purinergic P2; D000071557:beta-Arrestins; D020875:Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; D009020:Morphine; D002118:Calcium