Naunyn-Schmiedeberg's Archives of Pharmacology

, Volume 348, Issue 5, pp 504–508 | Cite as

The interaction of the mu-opioid receptor and G protein is altered after chronic morphine treatment in rats

  • Pao-Luh Tao
  • Chia-Rong Lee
  • Ping-Yee Law
  • Horace H. Loh
Original Articles
Received:
Accepted:

Summary

The interaction of µ-opioid receptors and G proteins after chronic morphine treatment was investigated in rats. Male Sprague-Dawley rats (200–260 g) were rendered tolerant to morphine by i.p. injections of increasing doses of morphine twice daily for 4 or 6 days. During this period, there was a time-dependent increase in the AD50 values for morphine to inhibit the tail-flick response. In addition, in vitro µ-opioid receptor binding to midbrain P2 membranes from these animals revealed that the ability of 10 μmol/l Gpp(NH)p (guanyl-5′-yl imidodiphosphate) to decrease [3H]DAMGO (TyrD-Ala-Gly-McPhe-Gly-ol) binding affinity, i.e., the ratio Kd(+Gpp(NH)p)/Kd(−Gpp(NH)p), decreased significantly from the control value of 3.68±0.40 to 2.37±0.35 after 6 days of morphine treatment (P<0.05).

The ability of DAMGO to stimulate low Km GTPase activity was also investigated. The EC50 significantly increased from 2.7± 1.1 × 10−8 mol/l in the control group to 10.8±1.5 × 10−8 mol/l after 4 days of morphine treatment and was further increased to 13.5±2.1 × 10−8 mol/l after 6 days of morphine treatment. The maximal stimulation by DAMGO decreased significantly from 18.0±1.7% to 12.8± 1.6% after 6 days of morphine treatment.

These results indicate that the interaction between µ-opioid receptors and G proteins had been altered after chronic morphine treatment.

Key words

Tolerance Morphine Opioid receptors G proteins GTPase Rat 
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References

  1. Barchfeld CC, Medzihradsky F (1984) Receptor-mediated stimulation of brain GTPase by opiates in normal and dependent rats. Biochem Biophys Res Commun 121:641–648Google Scholar
  2. Benovic JL, Pike LJ, Cerione RA, Staniszewski C, Yoshimasa T, Codina J, Caron MG, Lefkowitz RJ (1985) Phosphorylation of the mammalian β-adrenergic receptor by cyclic AMP-dependent protein kinase. J Biol Chem 260:7094–7101Google Scholar
  3. Benovic JL, Kuhn H, Weyand I, Codina J, Caron MT, Lefkowitz RJ (1987) Functional desensitization of the isolated β-adrenergic receptor by the β-adrenergic receptor kinase: potential role of an analog of the retinal protein arrestin (48-kDa protein). Proc Natl Acad Sci USA 84:8879–8882Google Scholar
  4. Blume AJ (1978) Interaction of ligands with the opiate receptors of brain membranes: regulation by ions and nucleotides. Proc Natl Acad Sci USA 75:1713–1717Google Scholar
  5. Burns DL, Hewlett EL, Moss J, Vaughan M (1983) Pertussis toxin inhibits enkephalin stimulation of GTPase of NG108–15 cells. J Biol Chem 258:1435–1438Google Scholar
  6. Cassel D, Selinger Z (1976) Catecholamine-stimulated GTPase activity in turkey erythrocyte membranes. Biochim Biophys Acta 452:538–551Google Scholar
  7. Chang K-J, Hazum E, Killian A, Cuatrecasas P (1981) Interactions of ligands with morphine and enkephalin receptors are differentially affected by guanine nucleotide. Mol Pharmacol 20:1–7Google Scholar
  8. Childers SR, Snyder SH (1980) Differential regulation by guanine nucleotides of opiate agonist and antagonist receptor interactions. J Neurochem 34:583–593Google Scholar
  9. D'Amour FE, Smith DL (1941) A method for determining loss of pain sensation. J Pharmacol Exp Ther 72:74–79Google Scholar
  10. Danks JA, Tortella FC, Bykov V, Jacobson AE, Rice KC, Holaday JW, Rothman RB (1988) Chronic administration of morphine and naltrexone up-regulate [3H][d-Ala2, d-Leu5]enkephalin binding sites by different mechanisms. Neuropharmacol 27:965–974Google Scholar
  11. Dixon WJ (1965) The up and down method for small samples. Am Stat Assoc J:967–978Google Scholar
  12. Frances B, Moisand C, Meunier J-C (1985) Na+ ions and Gpp(NH)p selectively inhibit agonist interactions at µ-and κ-opioid receptor sites in rabbit and guinea-pig cerebellum membranes. Eur J Pharmacol 117:223–232Google Scholar
  13. Hitzemann RJ, Hitzemann BA, Loh HH (1974) Binding of [3H] naloxone in the mouse brain: Effect of ions and tolerance development. Life Sci 14:2393–2404Google Scholar
  14. Holaday JW, Hitzemann RJ, Curell J, Tortella FC, Belenky GL (1982) Repeated electroconvulsive shock or chronic morphine treatment increase the number of [3H]-d-Ala2, d-Leu5-enkephalin in binding sites in rat brain membranes. Life Sci 31:2359–2362Google Scholar
  15. Hollt V, Dum J, Blasig J, Schubert P, Herz A (1975). Comparison of in vivo and in vitro parameters of opiate receptor binding in naive and tolerant/dependent rats. Life Sci 16:1823–1828Google Scholar
  16. Klee WA, Streaty RA (1974) Narcotic receptor sites in morphine-dependent rats. Nature 248:61–63Google Scholar
  17. Koski G, Klee WA (1981) Opiates inhibit adenylate cyclase by stimulating GTP hydrolysis. Proc Natl Acad Sci USA 78:4185–4189Google Scholar
  18. Law PY, Hom DS, Loh HH (1982) Loss of opiate receptor activity in Neuroblastoma x Glioma NG108–15 hybrid cells after chronic opiate treatment: a multiple step process. Mol Pharmacol 22:1–4Google Scholar
  19. Law PY, Hom DS, Loh HH (1983) Opiate receptor down-regulation and desensitization in Neuroblastoma x Glioma NG108–15 hybrid cells are two separate cellular adaptation processes. Mol Pharmacol 24:413–424Google Scholar
  20. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with Folin phenol reagent. J Biol Chem 193:265–275PubMedGoogle Scholar
  21. McPherson GA (1983) A practical computer-based approach to the analysis of radioligand binding experiments. Comput Programs Biomed 17:107–114Google Scholar
  22. Munson PJ, Rodbard D (1980) LIGAND: A versatile computerized approach for characterization of ligand binding systems. Anal Biochem 107:220–239Google Scholar
  23. Parenti M, Gazzotti G, Tirone F, Groppetti A (1983) Opiate tolerance and dependence is associated with a decreased activity of GTPase in rat striatal membranes. Life Sci 33, Sup I:345–348Google Scholar
  24. Pert CB, Snyder SH (1976) Opiate receptor binding enhancement by opiate administration in vivo. Biochem Pharmacol 25:847–853Google Scholar
  25. Puttfarcken PS, Werling LL, Cox BM (1988) 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 receptor. Mol Pharmacol 33:520–527Google Scholar
  26. Rogers NF, el-Fakahany E (1986) Morphine-induced opioid receptor down-regulation detected in intact adult rat brain cells. Eur J Pharmacol 124:221–230Google Scholar
  27. Rothman RB, Danks JA, Jacobson AE, Burke TR, Burke TR Jr, Rice KC, Tortella FC, Holaday JW (1986) Morphine tolerance increases mu-noncompetitive delta binding sites. Eur J Pharmacol 124: 113–119Google Scholar
  28. Rothman RB, Bykov V, Long JB, Brady LS, Jacobson AE, Rice KC, Holaday JW (1989) Chronic administration of morphine and naltrexone up-regulate mu-opioid binding sites labeled by [3H](d-Ala2, McPhe4, Gly-ol5)enkephalin: further evidence for two mu-binding sites. Eur J Pharmacol 160:71–82Google Scholar
  29. Simon EJ, Hiller JB (1978) In vitro studies on opiate receptors and their ligands. Fed Proc 37:141–146Google Scholar
  30. Sumner BE, Coombes JE, Pumford KM, Russell JA (1990) Opioid receptor subtypes in the supraoptic nucleus and posterior pituitary gland of morphine-tolerant rats. Neuroscience 37:635–645Google Scholar
  31. Tao P-L, Law PY, Loh HH (1987) Decrease in σ- and µ-opioid receptor binding capacity in rat brain after chronic etorphine treatment. J Pharmacol Exp Ther 240:809–816Google Scholar
  32. Tao P-L, Chang L-R, Law PY, Loh HH (1988) Decrease in σ-opioid receptor density in rat brain after chronic d-Ala2, d-Leu5-enkephalin treatment. Brain Res 462:313–320Google Scholar
  33. Tao P-L, Lee HY, Chang L-R, Loh HH (1990) Decrease in mu-opioid receptor binding capacity in rat brain after chronic PLO17 treatment. Brain Res 526:270–275Google Scholar
  34. Tao P-L, Tsai C-L, Chang L-R, Loh HH (1991) Chronic effect of (d-Pen2, d-Pen5)enkephalin on rat brain opioid receptors. Eur J Pharmacol 201:209–214Google Scholar
  35. Tempel A, Habas JE, Paredes W, Barr GA (1988) Morphine-induced downregulation of µ-opioid receptors in neonatal rat brain. Brain Res 469:129–133Google Scholar
  36. Vachon L, Costa T, Herz A (1985) Desensitization of opioid-stimulated GTPase in neuroblastoma x glioma hybrid cells. Biochem Biophys Res Commun 128:1342–1349Google Scholar
  37. Vachon L, Costa T, Herz A (1987) GTPase and adenylate cyclase desensitize at different rates in NG108–15 cells. Mol Pharmacol 31:159–168Google Scholar
  38. Werling LL, McMahon PN, Cox BM (1989) Selective changes in mu opioid receptor properties induced by chronic morphine exposure. Proc Natl Acad Sci USA 86:6393–6397Google Scholar
  39. Wilden U, Hall SW, Kuhn H (1986) Phosphodiesterase activation by photoexcited rhodopsin is quenched when rhodopsin is phosphorylated and binds the intrinsic 48-kDa protein of rod outer segments. Proc Natl Acad Sci USA 83:1174–1178Google Scholar
  40. Williams J, North RA (1983) Tolerance to opiates in locus coerulcus actions? Abstr Int Narc Res Conf p 49Google Scholar
  41. Wong C-S, Su Y-F, Watkins WD, Chang K-J (1992) Continuous intrathecal opioid treatment abolishes the regulatory effects of magnesium and guanine nucleotides on mu opioid receptor binding in rat spinal membranes. J Pharmacol Exp Ther 262:317–326Google Scholar

Copyright information

© Springer-Verlag 1993

Authors and Affiliations

  • Pao-Luh Tao
    • 1
  • Chia-Rong Lee
    • 1
  • Ping-Yee Law
    • 2
  • Horace H. Loh
    • 2
  1. 1.Department of PharmacologyNational Defense Medical CenterTaipei, TaiwanR.O.C.
  2. 2.Department of PharmacologyUniversity of MinnesotaMinneapolisUSA

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