Synergistic antinociceptive actions and tolerance development produced by morphine–fentanyl coadministration: Correlation with μ-opioid receptor internalization

2012 ◽  
Vol 674 (2-3) ◽  
pp. 239-247 ◽  
Author(s):  
Arturo Silva-Moreno ◽  
Claudia Gonzalez-Espinosa ◽  
Martha León-Olea ◽  
Silvia L. Cruz
Keyword(s):  
Opioid Receptor ◽  
Receptor Internalization ◽  
Tolerance Development ◽  
Μ Opioid Receptor

The μ opioid receptor: Cellular function and tolerance development

1994 ◽  
Vol 54 (1) ◽  
pp. 193-194 ◽  
Author(s):  
Anton Mestek ◽  
Yan Chen ◽  
Joyce H. Hurley ◽  
Leighan S. Bye ◽  
Andrew D. Campbell ◽  
...  
Keyword(s):  
Opioid Receptor ◽  
Cellular Function ◽  
Tolerance Development ◽  
Μ Opioid Receptor

scholarly journals Dissecting the roles of GRK2 and GRK3 in μ-opioid receptor internalization and β-arrestin2 recruitment using CRISPR/Cas9-edited HEK293 cells

2020 ◽  
Author(s):  
Thor C. Møller ◽  
Mie F. Pedersen ◽  
Jeffrey R. van Senten ◽  
Sofie D. Seiersen ◽  
Jesper M. Mathiesen ◽  
...  
Keyword(s):  
Opioid Receptor ◽  
G Protein Coupled Receptors ◽  
Receptor Internalization ◽  
Hek293 Cells ◽  
Molecule Inhibitor ◽  
Wide Range ◽  
Individual Contributions ◽  
Μ Opioid Receptor ◽  
G Protein Coupled ◽  
Target Effects

AbstractMost G protein-coupled receptors (GPCRs) recruit β-arrestins and internalize upon agonist stimulation. For the μ-opioid receptor (μ-OR), this process has been linked to development of opioid tolerance. GPCR kinases (GRKs), particularly GRK2 and GRK3, have been shown to be important for μ-OR recruitment of β-arrestin and internalization. However, the contribution of GRK2 and GRK3 to β-arrestin recruitment and receptor internalization, remain to be determined in their complete absence. Using CRISPR/Cas9-mediated genome editing we established HEK293 cells with knockout of GRK2, GRK3 or both to dissect their individual contributions in β-arrestin2 recruitment and μ-OR internalization upon stimulation with four different agonists. We showed that GRK2/3 removal reduced agonist-induced μ-OR internalization and β-arrestin2 recruitment substantially and we found GRK2 to be more important for these processes than GRK3. Furthermore, we observed a sustained and GRK2/3 independent component of β-arrestin2 recruitment to the plasma membrane upon μ-OR activation. Rescue expression experiments restored GRK2/3 functions. Inhibition of GRK2/3 using the small molecule inhibitor CMPD101 showed a high similarity between the genetic and pharmacological approaches, cross-validating the specificity of both. However, off-target effects were observed at high CMPD101 concentrations. These GRK2/3 KO cell lines should prove useful for a wide range of studies on GPCR function.

scholarly journals Dissecting the roles of GRK2 and GRK3 in μ-opioid receptor internalization and β-arrestin2 recruitment using CRISPR/Cas9-edited HEK293 cells

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Thor C. Møller ◽  
Mie F. Pedersen ◽  
Jeffrey R. van Senten ◽  
Sofie D. Seiersen ◽  
Jesper M. Mathiesen ◽  
...  
Keyword(s):  
Opioid Receptor ◽  
G Protein Coupled Receptors ◽  
Receptor Internalization ◽  
Hek293 Cells ◽  
Molecule Inhibitor ◽  
Wide Range ◽  
Individual Contributions ◽  
Μ Opioid Receptor ◽  
G Protein Coupled ◽  
Target Effects

Abstract Most G protein-coupled receptors (GPCRs) recruit β-arrestins and internalize upon agonist stimulation. For the μ-opioid receptor (μ-OR), this process has been linked to development of opioid tolerance. GPCR kinases (GRKs), particularly GRK2 and GRK3, have been shown to be important for μ-OR recruitment of β-arrestin and internalization. However, the contribution of GRK2 and GRK3 to β-arrestin recruitment and receptor internalization, remain to be determined in their complete absence. Using CRISPR/Cas9-mediated genome editing we established HEK293 cells with knockout of GRK2, GRK3 or both to dissect their individual contributions in β-arrestin2 recruitment and μ-OR internalization upon stimulation with four different agonists. We showed that GRK2/3 removal reduced agonist-induced μ-OR internalization and β-arrestin2 recruitment substantially and we found GRK2 to be more important for these processes than GRK3. Furthermore, we observed a sustained and GRK2/3 independent component of β-arrestin2 recruitment to the plasma membrane upon μ-OR activation. Rescue expression experiments restored GRK2/3 functions. Inhibition of GRK2/3 using the small molecule inhibitor CMPD101 showed a high similarity between the genetic and pharmacological approaches, cross-validating the specificity of both. However, off-target effects were observed at high CMPD101 concentrations. These GRK2/3 KO cell lines should prove useful for a wide range of studies on GPCR function.

μ-Opioid Receptor Internalization: Opiate Drugs Have Differential Effects on a Conserved Endocytic Mechanism In Vitro and in the Mammalian Brain

1998 ◽  
Vol 53 (3) ◽  
pp. 377-384 ◽  
Author(s):  
Duane E. Keith ◽  
Benito Anton ◽  
Stephen R. Murray ◽  
Paulette A. Zaki ◽  
Peter C. Chu ◽  
...  
Keyword(s):  
Opioid Receptor ◽  
Receptor Internalization ◽  
Mammalian Brain ◽  
Differential Effects ◽  
Opiate Drugs ◽  
Μ Opioid Receptor

Enhancement of Morphine Analgesic Effect with Induction of μ-Opioid Receptor Endocytosis in Rats

2006 ◽  
Vol 105 (3) ◽  
pp. 574-580 ◽  
Author(s):  
Tatsuya Hashimoto ◽  
Yoji Saito ◽  
Kazuo Yamada ◽  
Nobumasa Hara ◽  
Yumiko Kirihara ◽  
...  
Keyword(s):  
Opioid Receptor ◽  
Analgesic Effect ◽  
Antinociceptive Effect ◽  
Relative Activity ◽  
Receptor Internalization ◽  
Hot Plate Test ◽  
Analgesic Effects ◽  
Μ Opioid Receptor ◽  
Opiate Drug ◽  
Satisfactory Analgesia

Background Morphine can desensitize mu-opioid receptor (MOR), but it does not cause internalization of the receptor after binding. Acute desensitization of MOR impairs the efficiency of signaling, whereas the receptor internalization restores the cell responsiveness to the agonists. Thereby, the property of morphine may limit the analgesic effects of this opiate drug. It has been shown that [D-Ala2,MePhe4,Gly-ol5]enkephalin (DAMGO), a potent MOR agonist inducing the internalization, facilitates morphine to internalize MOR, suggesting that MOR agonists with low relative activity versus endocytosis (RAVE) values such as DAMGO can potentiate analgesic effects of morphine through stimulating MOR internalization. The authors examined whether the acute analgesic effect of morphine can be potentiated by low relative activity versus endocytosis agonists DAMGO and fentanyl. Methods Rats injected intrathecally with opioids were subjected to a hot plate test for antinociceptive effect. Immunostained spinal dorsal horn was analyzed by confocal microscopy. Results Fentanyl induced MOR internalization to a lesser extent than DAMGO at equianalgesic doses. Coadministration of fentanyl promoted morphine-induced MOR internalization. The analgesic effect of morphine was greatly potentiated together with decrease in the relative activity versus endocytosis value when MOR internalization was induced by coadministration of a subanalgesic dose of DAMGO or fentanyl. In contrast, the combination of DAMGO and fentanyl increased neither the analgesic effect nor the internalization of MOR. Conclusions The results suggest that the coadministration of morphine with MOR-internalizing agonist is clinically applicable to develop successful pain-management regimens to achieve satisfactory analgesia using less morphine.

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