Protein Kinase C-Mediated Inhibition of m-Opioid Receptor Internalization and Its Involvement in the Development of Acute Tolerance to Peripheral m-Agonist Analgesia

We investigated the role of protein kinase C (PKC) in cell m-opioid receptor (MOR) internalization and MOR-mediated acute tolerance in vivo. When Chinese hamster ovary cells expressing MOR were exposed to [D-Ala,MePhe,Gly-ol]enkephalin (DAMGO), receptor internalization was observed at 30 min. Incubation with morphine failed to induce receptor internalization. When calphostin C, a PKC inhibitor, was added, receptor internalization was observed as early as 10 min after morphine stimulation. The MOR internalization induced by DAMGO or morphine in the presence of calphostin C was dynamin dependent, because it was abolished 2 d after pretreatment with recombinant adenovirus to express a dominant interfering dynamin mutant (K44A/dynamin adenovirus). On the other hand, in a peripheral nociception test in mice, the nociceptive flexor response after intraplantar injection (i.pl.) of bradykinin was markedly inhibited by DAMGO (i.pl.). DAMGO analgesia was not affected by 2 hr prior injection (i.pl.) of DAMGO. Marked acute tolerance was observed after pretreatment with dynamin antisense oligodeoxynucleotide or K44A/dynamin adenovirus. The DAMGO-induced acute tolerance under such pretreatments was inhibited by calphostin C. Together, these findings suggest that PKC desensitizes MOR or has a role in the development of acute tolerance through MOR by inhibiting internalization mechanisms as a resensitization process.

[1]  H. Ueda,et al.  Protein kinase C-mediated acute tolerance to peripheral mu-opioid analgesia in the bradykinin-nociception test in mice. , 2000, The Journal of pharmacology and experimental therapeutics.

[2]  G. King,et al.  Identification of PKC-isoform-specific biological actions using pharmacological approaches. , 2000, Trends in pharmacological sciences.

[3]  L. Jan,et al.  Functional Dissociation of μ Opioid Receptor Signaling and Endocytosis Implications for the Biology of Opiate Tolerance and Addiction , 1999, Neuron.

[4]  C. Sternini,et al.  Activation and internalization of the μ-opioid receptor by the newly discovered endogenous agonists, endomorphin-1 and endomorphin-2 †† These authors contributed equally to this work. †† , 1999, Neuroscience.

[5]  M. Bünemann,et al.  G‐protein coupled receptor kinases as modulators of G‐protein signalling , 1999, The Journal of physiology.

[6]  H. Loh,et al.  The Absence of a Direct Correlation between the Loss of [d-Ala2,MePhe4,Gly5-ol]Enkephalin Inhibition of Adenylyl Cyclase Activity and Agonist-induced μ-Opioid Receptor Phosphorylation* , 1999, The Journal of Biological Chemistry.

[7]  H Weinstein,et al.  Functional role of the spatial proximity of Asp114(2.50) in TMH 2 and Asn332(7.49) in TMH 7 of the μ opioid receptor , 1999, FEBS letters.

[8]  H. Ueda.  In vivo molecular signal transduction of peripheral mechanisms of pain. , 1999, Japanese journal of pharmacology.

[9]  C. Chavkin,et al.  Agonist induced homologous desensitization of mu-opioid receptors mediated by G protein-coupled receptor kinases is dependent on agonist efficacy. , 1998, Molecular pharmacology.

[10]  J. Pessin,et al.  Expression of a Dominant Interfering Dynamin Mutant in 3T3L1 Adipocytes Inhibits GLUT4 Endocytosis without Affecting Insulin Signaling* , 1998, The Journal of Biological Chemistry.

[11]  H. Ueda,et al.  Nociceptin/orphanin FQ-induced nociceptive responses through substance P release from peripheral nerve endings in mice. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[12]  M. Caron,et al.  Role for G protein-coupled receptor kinase in agonist-specific regulation of mu-opioid receptor responsiveness. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[13]  M. Comb,et al.  A Mitogen-activated Protein Kinase Pathway Is Required for μ-Opioid Receptor Desensitization* , 1998, The Journal of Biological Chemistry.

[14]  D. Massotte,et al.  Desensitization of the δ‐Opioid Receptor Correlates with Its Phosphorylation in SK‐N‐BE Cells: Involvement of a G Protein‐Coupled Receptor Kinase , 1998, Journal of neurochemistry.

[15]  D. Lissin,et al.  mu-Opioid receptor internalization: opiate drugs have differential effects on a conserved endocytic mechanism in vitro and in the mammalian brain. , 1998, Molecular pharmacology.

[16]  H. Loh,et al.  Role of carboxyl terminus of μ- and δ-opioid receptor in agonist -induced down-regulation , 1998 .

[17]  W. Sadee,et al.  Specific G protein activation and mu-opioid receptor internalization caused by morphine, DAMGO and endomorphin I. , 1998, European journal of pharmacology.

[18]  M. Caron,et al.  A central role for beta-arrestins and clathrin-coated vesicle-mediated endocytosis in beta2-adrenergic receptor resensitization. Differential regulation of receptor resensitization in two distinct cell types. , 1997, The Journal of biological chemistry.

[19]  S. Schulz,et al.  Immunolocalization of two mu-opioid receptor isoforms (MOR1 and MOR1B) in the rat central nervous system , 1997, Neuroscience.

[20]  B. O'dowd,et al.  Agonist-induced Desensitization of the μ Opioid Receptor Is Determined by Threonine 394 Preceded by Acidic Amino Acids in the COOH-terminal Tail* , 1997, The Journal of Biological Chemistry.

[21]  T. Kroslak,et al.  Site Mutation in the Rat μ‐Opioid Receptor Demonstrates the Involvement of Calcium/Calmodulin‐Dependent Protein Kinase II in Agonist‐Mediated Desensitization , 1997, Journal of neurochemistry.

[22]  A. Hornia,et al.  Tumor promotion by depleting cells of protein kinase C delta , 1997, Molecular and cellular biology.

[23]  Julie A. Pitcher,et al.  The Role of Sequestration in G Protein-coupled Receptor Resensitization , 1997, The Journal of Biological Chemistry.

[24]  M. von Zastrow,et al.  Agonist-selective endocytosis of mu opioid receptor by neurons in vivo. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[25]  Christopher J. Evans,et al.  Morphine Activates Opioid Receptors without Causing Their Rapid Internalization* , 1996, The Journal of Biological Chemistry.

[26]  M. Caron,et al.  Dynamin and β-Arrestin Reveal Distinct Mechanisms for G Protein-coupled Receptor Internalization* , 1996, The Journal of Biological Chemistry.

[27]  D. Lambert,et al.  The stimulatory effects of opioids and their possible role in the development of tolerance. , 1996, Trends in pharmacological sciences.

[28]  F. F. Weight,et al.  Differential Opiate Receptor Phosphorylation and Desensitization Induced by Agonists and Phorbol Esters (*) , 1996, The Journal of Biological Chemistry.

[29]  M. Lohse,et al.  Protein kinase cross-talk: membrane targeting of the beta-adrenergic receptor kinase by protein kinase C. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[30]  H. Ueda,et al.  Protein kinase C involvement in homologous desensitization of delta- opioid receptor coupled to Gi1-phospholipase C activation in Xenopus oocytes , 1995, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[31]  H. Ueda,et al.  Protein kinase C inhibitor potentiates the agonist-induced GTPase activity in COS cell membranes expressing δ-opioid receptor , 1995 .

[32]  M. Narita,et al.  Inhibition of protein kinase C, but not of protein kinase A, blocks the development of acute antinociceptive tolerance to an intrathecally administered mu-opioid receptor agonist in the mouse. , 1995, European journal of pharmacology.

[33]  L. Toll.  Intact cell binding and the relation to opioid activities in SH-SY5Y cells. , 1995, The Journal of pharmacology and experimental therapeutics.

[34]  H. Schulman,et al.  The human mu opioid receptor: modulation of functional desensitization by calcium/calmodulin-dependent protein kinase and protein kinase C , 1995, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[35]  T. Nakagawa,et al.  DAMGO, a μ‐opioid receptor selective agonist, distinguishes between μ‐ and δ‐opioid receptors around their first extracellular loops , 1995 .

[36]  H. Ueda,et al.  Species and age-dependent differences of functional coupling between opioid δ-receptor and G-proteins and possible involvement of protein kinase C in striatal membranes , 1994, Neuroscience Letters.

[37]  K. Wenzel-Seifert,et al.  N-protein kinase C isoenzymes may be involved in the regulation of various neutrophil functions. , 1994, Biochemical and biophysical research communications.

[38]  Y. Chen,et al.  Differential regulation by cAMP-dependent protein kinase and protein kinase C of the mu opioid receptor coupling to a G protein-activated K+ channel. , 1994, The Journal of biological chemistry.

[39]  W. Janzen,et al.  New hexahydroxybiphenyl derivatives as inhibitors of protein kinase C. , 1994, Journal of medicinal chemistry.

[40]  M. Nishi,et al.  Primary structures and expression from cDNAs of rat opioid receptor δ‐and μ‐subtypes , 1993 .

[41]  M. Hagiwara,et al.  The newly synthesized selective Ca2+/calmodulin dependent protein kinase II inhibitor KN-93 reduces dopamine contents in PC12h cells. , 1991, Biochemical and biophysical research communications.

[42]  T. Katada,et al.  Phosphorylated μ-opioid receptor purified from rat brains lacks functional coupling with Gi1, a GTP-binding protein in reconstituted lipid vesicles , 1990, Neuroscience Letters.

[43]  C. Murakata,et al.  K-252 compounds, novel and potent inhibitors of protein kinase C and cyclic nucleotide-dependent protein kinases. , 1987, Biochemical and biophysical research communications.

[44]  M. Zimmermann,et al.  Ethical guidelines for investigations of experimental pain in conscious animals , 1983, Pain.

[45]  G. Wilcox,et al.  Intrathecal morphine in mice: a new technique. , 1980, European journal of pharmacology.

[46]  H. Loh,et al.  Molecular mechanisms and regulation of opioid receptor signaling. , 2000, Annual review of pharmacology and toxicology.

[47]  C. Chavkin,et al.  Mu and delta opioid receptors are differentially desensitized by the coexpression of beta-adrenergic receptor kinase 2 and beta-arrestin 2 in xenopus oocytes. , 1997, The Journal of biological chemistry.

[48]  T. Nakagawa,et al.  DAMGO, a mu-opioid receptor selective agonist, distinguishes between mu- and delta-opioid receptors around their first extracellular loops. , 1995, FEBS letters.

[49]  C. Newgard,et al.  Use of recombinant adenovirus for metabolic engineering of mammalian cells. , 1994, Methods in cell biology.