Abstract
Morphine is a powerful pain reliever, but also a potent inducer of tolerance and dependence. The development of opiate tolerance occurs on continued use of the drug such that the amount of drug required to elicit pain relief must be increased to compensate for diminished responsiveness1,2,3. In many systems, decreased responsiveness to agonists has been correlated with the desensitization of G-protein-coupled receptors. In vitro evidence indicates that this process involves phosphorylation of G-protein-coupled receptors and subsequent binding of regulatory proteins called β-arrestins4,5. Using a knockout mouse lacking β-arrestin-2 (βarr2-/-), we have assessed the contribution of desensitization of the μ-opioid receptor to the development of morphine antinociceptive tolerance and the subsequent onset of physical dependence. Here we show that in mice lacking β-arrestin-2, desensitization of the μ-opioid receptor does not occur after chronic morphine treatment, and that these animals fail to develop antinociceptive tolerance. However, the deletion of β-arrestin-2 does not prevent the chronic morphine-induced upregulation of adenylyl cyclase activity, a cellular marker of dependence, and the mutant mice still become physically dependent on the drug.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 51 print issues and online access
$199.00 per year
only $3.90 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
References
Nestler, E. J. Under siege: The brain on opiates. Neuron 16, 897–900 (1996).
Nestler, E. J. & Aghajanian, G. K. Molecular and cellular basis of addiction. Science 278, 58–63 (1997).
Koob, G. F., Sanna, P. P. & Bloom, F. E. Neuroscience of addiction. Neuron 21, 467–476 (1998).
Ferguson, S. S., Barak, L. S., Zhang, J. & Caron, M. G. G-protein-coupled receptor regulation: role of G-protein-coupled receptor kinases and arrestins. Can. J. Physiol. Pharmacol. 74, 1095–1110 (1996).
Pitcher, J. A., Freedman, N. J. & Lefkowitz, R. J. G protein-coupled receptor kinases. Annu. Rev. Biochem. 67, 653–692 (1998).
Bohn, L. M. et al. Enhanced morphine analgesia in mice lacking β-arrestin 2. Science 286, 2495–2498 (1999).
Matthes, H. W. et al. Loss of morphine-induced analgesia, reward effect and withdrawal symptoms in mice lacking the µ-opioid-receptor gene. Nature 383, 819–823 (1996).
Sora, I. et al. Opiate receptor knockout mice define µ receptor roles in endogenous nociceptive responses and morphine-induced analgesia. Proc. Natl Acad. Sci. USA 94, 1544–1549 (1997).
Kieffer, B. L. Opioids: first lessons from knockout mice. Trends Pharmacol. Sci. 20, 19–26 (1999).
Way, E. L., Loh, H. H. & Shen, F. H. Simultaneous quantitative assessment of morphine tolerance and physical dependence. J. Pharmacol. Exp. Ther. 167, 1–8 (1969).
Brase, D. A., Loh, H. H. & Way, E. L. Comparison of the effects of morphine on locomotor activity, analgesia and primary and protracted physical dependence in six mouse strains. J. Pharmacol. Exp. Ther. 201, 368–374 (1977).
Schulteis, G., Markou, A., Gold, L. H., Stinus, L. & Koob, G. F. Relative sensitivity to naloxone of multiple indices of opiate withdrawal: a quantitative dose–response analysis. J. Pharmacol. Exp. Ther. 271, 1391–1398 (1994).
Jolas, T., Nestler, E. J. & Aghajanian, G. K. Chronic morphine increases GABA tone on serotonergic neurons of the dorsal raphe nucleus: association with an up-regulation of the cyclic AMP pathway. Neuroscience 95, 433–443 (2000).
Cox, B. M. in Opioids in Pain Control: Basic and Clinical Aspects (ed. C. Stein) 109–130 (Cambridge Univ. Press, New York, 1999).
Collier, H. O. Cellular site of opiate dependence. Nature 283, 625–629 (1980).
Avidor-Reiss, T. et al. Adenylylcyclase supersensitization in µ-opioid receptor-transfected Chinese hamster ovary cells following chronic opioid treatment. J. Biol. Chem. 270, 29732–29738 (1995).
Puttfarcken, P. S., Werling, L. L. & Cox, B. M. Effects of chronic morphine exposure on opioid inhibition of adenylyl cyclase in 7315c cell membranes: a useful model for the study of tolerance at µ opioid receptors. Mol. Pharmacol. 33, 520–527 (1988).
Breivogel, C. S., Selley, D. E. & Childers, S. R. Acute and chronic effects of opioids on δ and µ receptor activation of G proteins in NG108-15 and SK-N-SH cell membranes. J. Neurochem. 68, 1462–1472 (1997).
Elliott, J., Guo, L. & Traynor, J. R. Tolerance to µ-opioid agonists in human neuroblastoma SH-SY5Y cells as determined by changes in guanosine-5′-O-(3-[35S]-thio)triphosphate binding. Br. J. Pharmacol. 121, 1422–1428 (1997).
Sim, L. J., Hampson, R. E., Deadwyler, S. A. & Childers, S. R. Effects of chronic treatment with Δ9-tetrahydrocannabinol on cannabinoid-stimulated [35S]GTPγS autoradiography in rat brain. J. Neurosci. 16, 8057–8066 (1996).
Noble, F. & Cox, B. M. Differential desensitization of µ- and δ-opioid receptors in selected neural pathways following chronic morphine treatment. Br. J. Pharmacol. 117, 161–169 (1996).
Kovoor, A., Nappey, V., Kieffer, B. L. & Chavkin, C. µ and δ opioid receptors are differentially desensitized by the coexpression of β-adrenergic receptor kinase 2 and β-arrestin 2 in xenopus oocytes. J. Biol. Chem. 272, 27605–27611 (1997).
Zhang, J. et al. Role for G protein-coupled receptor kinase in agonist-specific regulation of µ-opioid receptor responsiveness. Proc. Natl Acad. Sci. USA 95, 7157–7162 (1998).
Whistler, J. L. & von Zastrow, M. Morphine-activated opioid receptors elude desensitization by β-arrestin. Proc. Natl Acad. Sci. USA 95, 9914–9919 (1998).
Lane-Ladd, S. B. et al. CREB (cAMP response element-binding protein) in the locus coeruleus: biochemical, physiological, and behavioral evidence for a role in opiate dependence. J. Neurosci. 17, 7890–7901 (1997).
Fairbanks, C. A. & Wilcox, G. L. Spinal antinociceptive synergism between morphine and clonidine persists in mice made acutely or chronically tolerant to morphine. J. Pharmacol. Exp. Ther. 288, 1107–1116 (1999).
Zhu, Y. et al. Retention of supraspinal delta-like analgesia and loss of morphine tolerance in delta opioid receptor knockout mice. Neuron 24, 243–252 (1999).
Gainetdinov, R. R. et al. Muscarinic supersensitivity and impaired receptor desensitization in G protein-coupled receptor kinase 5-deficient mice. Neuron 24, 1029–1036 (1999).
Hausdorff, W. P., Hnatowich, M., O'Dowd, B. F., Caron, M. G. & Lefkowitz, R. J. A mutation of the β 2-adrenergic receptor impairs agonist activation of adenylyl cyclase without affecting high affinity agonist binding. Distinct molecular determinants of the receptor are involved in physical coupling to and functional activation of Gs. J. Biol. Chem. 265, 1388–1393 (1990).
Zhang, J., Barak, L. S., Winkler, K. E., Caron, M. G. & Ferguson, S. S. A central role for β-arrestins and clathrin-coated vesicle-mediated endocytosis in β2-adrenergic receptor resensitization. Differential regulation of receptor resensitization in two distinct cell types. J. Biol. Chem. 272, 27005–27014 (1997).
Acknowledgements
We thank S. Suter for care and genotyping of animals. R.R.G. is a visiting scientist from the Institute of Pharmacology, Russian Academy of Medical Sciences, Baltiyskaya 8, 125315 Moscow, Russia. M.G.C. and R.J.L. are Investigators of the Howard Hughes Medical Institute.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Bohn, L., Gainetdinov, R., Lin, FT. et al. μ-Opioid receptor desensitization by β-arrestin-2 determines morphine tolerance but not dependence. Nature 408, 720–723 (2000). https://doi.org/10.1038/35047086
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/35047086
This article is cited by
-
The multifaceted functions of β-arrestins and their therapeutic potential in neurodegenerative diseases
Experimental & Molecular Medicine (2024)
-
Genetic mouse models in opioid research: current status and future directions
Journal of Neural Transmission (2024)
-
Heroin and its metabolites: relevance to heroin use disorder
Translational Psychiatry (2023)
-
Essential role of P-glycoprotein in the mechanism of action of oliceridine
Neuropsychopharmacology (2023)
-
Molecular mechanisms of morphine tolerance and dependence; novel insights and future perspectives
Molecular and Cellular Biochemistry (2023)
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.