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Interaction of calcium with3H-morphine binding to synaptosomes of the guinea-pig ileum

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Summary

Specific binding sites for3H-morphine were assayed in subcellular fractions of a crude mitochondrial P2 pellet from the guinea-pig ileum longitudinal muscle-myenteric plexus, prepared by discontinuous sucrose-gradient fractionation. The highest specific binding (ca. 100 fmol/mg protein) was obtained in the fraction containing synaptosomes, as examined by electron microscopy. A synaptosomal fraction prepared from guinea-pig brain had comparable specific binding (ca. 130 fmol/mg) to that of the ileal synaptosomal fraction. Addition of calcium (10 mM) to the binding assay medium resulted in a marked decrease in particular of the specific3H-morphine binding. Detailed analysis of the specific3H-morphine binding in the synaptosomal fraction of the ileum revealed that 1) a saturable component of specific opiate binding was present between 0.34 and 21.74 nM of3H-morphine; 2) in the presence of 3 and 10 nM of calcium similar decreases of specific3H-morphine binding were obtained, indicating that this binding was maximally inhibited already by 3 mM of calcium; 3) both in the absence and presence of calcium theK D of specific3H-morphine binding was about 38 nM, indicating a non-competitive nature of the calcium inhibition; 4) addition of magnesium exhibited a similar effect as that of calcium, although magnesium appeared to be less potent than calcium in this respect. The data are discussed in the context of previously observed calcium effects on opioid actions in the electrically stimulated guinea-pig ileum bioassay and may contribute to the evidence that the interaction of calcium and opioids on the stimulus-release coupling mechanism at the neuromuscular junction of the guinea-pig ileum is occurring beyond opioid receptor activation as well.

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References

  • Abood LG, Takeda F, Salem N Jr (1977) Role of phosphatidyl serine in the opiate receptor. In: Blum K (ed) Alcohol and opiates: neurochemical and behavioral mechanisms. Academic Press, New York, pp 283–297

    Google Scholar 

  • Chapman DB, Way EL (1980) Metal ion interactions with opiates. Annu Rev Pharmacol Toxicol 20:553–579

    Google Scholar 

  • Creese I, Snyder SH (1975) Receptor binding and pharmacological activity of opiates in the guinea-pig intestine. J Pharmacol Exp Ther 194:205–219

    Google Scholar 

  • Ehrenpreis S (1975) Determination of actions of narcotic analgesics and their antagonists on electrically stimulated guinea-pig ileum. In: Ehrenpreis S, Neidle A (eds) Methods in narcotics research, vol. 5. Dekker, New York, pp 111–125

    Google Scholar 

  • Elde R, Hökfelt T, Johansson O, Terenius L (1976) Immuno-histochemical studies using antibodies to Leu-Enk: initial observations on the nervous system of the rat. Neuroscience 1:349–351

    Google Scholar 

  • Fischel SV, Medzihradsky F (1980) Scatchard analysis of opiate receptor binding. In: Way EL (ed) Endogenous and exogenous opiate agonists and antagonists. Pergamon Press, New York, pp 91–94

    Google Scholar 

  • Gabella G (1972) Fine structure of the myenteric plexus in the guinea-pig ileum. J Anat 111:69–97

    Google Scholar 

  • Giao JM, Rico T, Matos Da Costa PS (1978) A quantitative study of calcium-magnesium antagonism in isolated strips of guinea pig ileum. Eur J Pharmacol 48:411–419

    Google Scholar 

  • Guerrero-Munoz F, Guerrero ML, Way EL (1978) The effect of acute and chronic morphine administration on synaptosomal calcium uptake. In: van Ree JM, Terenius L (eds) Characteristics and function of opioids. Elsevier/North-Holland, Amsterdam, pp 311–321

    Google Scholar 

  • Harris RA, Loh HH, Way EL (1975) Effects of divalent cations, cation chelators and an ionophore on morphine analgesia and tolerance. J Pharmacol Exp Ther 195:488–498

    Google Scholar 

  • Hitzemann RJ, Hitzemann BA, Loh HH (1974) Binding of3H-naloxone in the mouse brain: effect of ions and tolerance development. Life Sci 14:2393–2404

    Google Scholar 

  • Kosterlitz HW, Waterfield AA (1975) In vitro models in the study of structure-activity relationships of narcotic analgetics. Annu Rev Pharmacol 15:29–47

    Google Scholar 

  • Kosterlitz HW, Leslie FM (1978) Comparison of the receptor binding characteristics of opiate agonists interacting with μ and ϰ-receptors. Brit J Pharmacol 64:607–614

    Google Scholar 

  • Labella F, Havlicek V, Pinsky C (1978) Opiates, steroids, Ca2+ antagonists: Comparison of central excitatory effects. In: van Ree JM, Terenius L (eds) Characteristics and function of opioids. Elsevier/North-Holland, Amsterdam, pp 361–362

    Google Scholar 

  • Law PY, Houghton RA, Loh HH, Li CH (1980) Characterization of a high affinity3H-βh-endorphin receptor in rat brain crude synaptosomal fraction. In: Way EL (ed) Endogenous and exogenous opiate agonists and antagonists. Pergamon Press, New York, pp 225–228

    Google Scholar 

  • Lord JAH, Waterfield AA, Hughes J, Kosterlitz HW (1977) Endogenous opioid peptides: multiple agonists and receptors. Nature 267:495–497

    Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275

    Google Scholar 

  • Opmeer FA, van Ree JM (1978) Quantification of the in vitro induced tolerance to morphine of the isolated guinea pig ileum. Neuropharmacology 17:887–890

    Google Scholar 

  • Opmeer FA, van Ree JM (1979) Competitive antagonism of morphine action in vitro by calcium. Eur J Pharmacol 53:395–397

    Google Scholar 

  • Opmeer FA, van Ree JM (1980) Differential involvement of calcium in acute and chronic opioid action in the guinea-pig ileum in vitro. J Pharmacol Exp Ther 213:188–195

    Google Scholar 

  • Pasternak GW, Snowman AM, Snyder SH (1975) Selective enhancement of [3H]-opiate agonist binding by divalent cations. Mol Pharmacol 11:735–744

    Google Scholar 

  • Paton WDM (1957) The action of morphine and related substances on contraction and on acetylcholine output of coaxially stimulated guinea-pig ileum. Brit J Pharmacol Chemother 12:119–127

    Google Scholar 

  • Pert CB, Snyder SH (1973) Opiate receptor: Demonstration in nervous tissue. Science 179:1011–1014

    Google Scholar 

  • Ross DH, Cardenas HL (1977) Levorphanol inhibition of Ca2+ binding to synaptic membranes in vitro. Life Sci 20:1455–1462

    Google Scholar 

  • Snyder SH, Childers SR (1979) Opiate receptors and opioid peptides. Annu Rev Neurosci 2:35–64

    Google Scholar 

  • Terenius L (1973) Stereospecific uptake of narcotic analgesics by a subcellular fraction of the guinea-pig ileum. Uppsala J Med Sci 78:150–152

    Google Scholar 

  • Waterfield AA, Kosterlitz HW (1975) Stereospecific increase by narcotic antagonist of evoked acetylcholine output in guinea-pig ileum. Life Sci 16:1787–1792

    Google Scholar 

  • Way EL (1978) Common and selective mechanisms in drug dependence. In: Fishman J (ded) The bases of addiction. Dahlem Konferenzen, Berlin, pp 333–352

  • Whittaker VP (1965) The application of subcellular fractionation techniques to the study of brain function. Prog Biophys Mol Biol 15:39–96

    Google Scholar 

  • Whittaker VP, Barker LA (1972) The subcellular fractionation of brain tissue with special reference to the preparation of synaptosomes and their component organelles. In: Fried R (ed). Methods of neurochemistry, vol 2. Dekker, New York, pp 1–52

    Google Scholar 

  • Yamamoto H, Harris RA, Loh HH, Way EL (1978) Effects of acute and chronic morphine treatments on calcium localization and binding in brain. J Pharmacol Exp Ther 205:255–264

    Google Scholar 

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Opmeer, F.A., Witter, A. & van Ree, J.M. Interaction of calcium with3H-morphine binding to synaptosomes of the guinea-pig ileum. Naunyn-Schmiedeberg's Arch. Pharmacol. 321, 1–6 (1982). https://doi.org/10.1007/BF00586340

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  • DOI: https://doi.org/10.1007/BF00586340

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