Summary
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1.
Pairs of smooth muscles isolated from the nictitating membrane of the cat were incubated with 1.2 ml of Krebs' solution containing 10 ng/ml of 3H-(±)-noradrenaline for 7.5 min (in the presence of U-0521 to inhibit COMT). Removal of the amine from the bath as well as the appearance of deaminated 3H-catechols in the bath were measured.
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2.
Pretreatment with reserpine did not affect the rate of removal, while increasing the rate of deamination. The ability of the muscles to retain exogenous amine for one hour was reduced to 12% of normal.
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3.
A certain fraction of the total production of deaminated 3H-catechols escaped into the medium. For any given duration of incubation this fraction was independent of the concentration of noradrenaline in the medium. On repeated incubation the fraction remained constant. Therefore, reliable estimates of the rate of deamination were obtained with repeated incubations of the same muscle.
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4.
Sympathetic denervation and/or cocaine revealed that 60% of removal (of which 10% are due to dilution) and 25% of deamination are extraneuronal.
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5.
For incubations of 7.5 min measured rates of deamination represent initial rates, measured rates of removal do not.
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6.
Unlabelled (−)- and (+)-noradrenaline were equipotent (ID50=about 1 μM) in inhibiting the deamination of 10 ng/ml of 3H-(±)-noradrenaline. This inhibitory effect must be exerted on neuronal deamination, since extraneuronal deamination (in denervated muscles) was not affected by the addition of unlabelled isomers.
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7.
It is proposed that, under these experimental conditions, neuronal unptake is the rate limiting step for neuronal deamination, and that neuronal uptake in the cat's nictitating membrane lacks stereoselectivity.
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References
Blaschko, H., Richter, D., Schlossmann, H.: The oxidation of adrenaline and other amines. Biochem. J. 31, 2187–2196 (1937).
Burn, J. H.: Practical Pharmacology. Oxford: Blackwell 1952.
Cervoni, P.: Monoamine oxidase activity of the cat nictitating membrane and superior cervical ganglion under various experimental conditions. Biochem. Pharmacol., 18, 1427–1433 (1969).
Coyle, J. T., Snyder, S. H.: Catecholamine uptake by synaptosomes in homogenates of rat brain: stereospecificity in different areas. J. Pharmacol. exp. Ther. 170, 221–231 (1969).
Draskóczy, P. R., Trendelenburg, U.: The uptake of l- and d-norepinephrine by the isolated perfused rabbit heart in relation to the stereospecificity of the sensitizing action of cocaine. J. Pharmacol. exp. Ther. 159, 66–73 (1968).
Draskóczy, P. R., Trendelenburg, U.: Intraneuronal and extraneuronal accumulation of sympathomimetic amines in the isolated nictitating membrane of the cat. J. Pharmacol. exp. Ther. 174, 290–306 (1970).
Esterhuizen, A. C., Graham, J. D. P., Lever, J. D., Spriggs, T. L. B.: Catecholamine and acetylcholinesterase distribution in relation to noradrenaline release. An enzyme histochemical and autoradiographic study on the innervation of the cat nictitating membrane. Brit. J. Pharmacol. 32, 46–56 (1968).
Euler, U. S. v., Lishajko, F.: Reuptake and net uptake of noradrenaline in adrenergic nerve granules with a note on the affinity for l- and d-isomers. Acta physiol. scand. 71, 151–162 (1967).
Finney, D. J.: Probit Analysis, 2nd ed. Cambridge: University Press 1964.
Giachetti, A., Shore, P. A.: Optical specificity of monoamine oxidase. Life Sci. 5, 1373–1378 (1966).
Graefe, K. H., Bönisch, H., Trendelenburg, U.: Time-dependent changes in neuronal uptake of noradrenaline after pretreatment with pargyline and/or reserpine. Naunyn-Schmiedebergs Arch. Pharmak. 271, 1–28 (1971).
Graefe, K. H., Trendelenburg, U.: The effect of cocaine on uptake of and sensitivity to noradrenaline in isolated nictitating membranes before and after storage in the cold. Naunyn-Schmiedebergs Arch. Pharmak. 267, 383–398 (1970).
Häggendal, J.: On the use of strong exchange resins for determinations of small amounts of catechol amines. Scandinav. J. clin. Lab. Invest. 14, 537–544 (1962).
Iversen, L. L.: The uptake of noradrenaline by the isolated perfused rat heart. Brit. J. Pharmacol. 21, 523–537 (1963).
Iversen, L. L.: Inhibition of noradrenaline uptake by sympathomimetic amines. J. Pharm. Pharmacol. 16, 435–436 (1964).
Iversen, L. L.: The uptake of adrenaline by the rat isolated heart. Brit. J. Pharmacol. 24, 387–394 (1965).
Iversen, L. L.: Characteristics of noradrenaline uptake in the iris/ciliary body and other peripheral tissues of the rat. Naunyn-Schmiedebergs Arch. Pharmak. exp. Path. 259, 179 (1968).
Iversen, L. L., Glowinski, J., Axelrod, J.: The uptake and storage of H3-norepinephrine in the reserpine-pretreated rat heart. J. Pharmacol. exp. Ther. 150, 173–183 (1965).
Iversen, L. L., Jarrott, B., Simmonds, M. A.: Differences in the uptake and metabolism of (+)- and (−)-noradrenaline. Brit. J. Pharmacol. 43, 845–855 (1971).
Jarrott, B.: Uptake and metabolism of catecholamines in the perfused hearts of different species. Brit. J. Pharmacol. 38, 810–821 (1970).
Jarrott, B., Langer, S. Z.: Changes in monoamine oxidase and catechol-O-methyl transferase activities after denervation of the nictitating membrane of the cat. J. Physiol. (Lond.) 212, 549–559 (1971).
Kopin, I. J., Hertting, G., Gordon, E. K.: Fate of norepinephrine-H3 in the isolated perfused rat heart. J. Pharmacol. exp. Ther. 138, 34–40 (1962).
Langer, S. Z., Trendelenburg, U.: The effect of a saturable uptake mechanism on the slopes of dose-response curves for sympathomimetic amines and on the shifts of dose-response curves produced by a competitive antagonist. J. Pharmacol. exp. Ther. 167, 117–142 (1969).
Lindmar, R., Muscholl, E.: Die Wirkung von Pharmaka auf die Elimination von Noradrenalin aus der Perfusionsflüssigkeit und die Noradrenalinaufnahme in das isolierte Herz. Naunyn-Schmiedebergs Arch. exp. Path. Pharmak. 247, 469–492 (1964).
Maickel, R. P., Beaven, M. A., Brodie, B. B.: Implications of uptake and storage of norepinephrine by sympathetic nerve endings. Life Sci. 2, 953–958 (1963).
Sachs, Ch.: Noradrenaline uptake mechanism in the mouse atrium. A biochemical and histochemical study. Acta physiol. scand. 79, suppl. 341, 1–67 (1970).
Trendelenburg, U.: Supersensitivity by cocaine to dextrorotatory isomers of norepinephrine and epinephrine. J. Pharmacol. exp. Ther. 148, 329–338 (1965).
Trendelenburg, U., Höhn, D., Graefe, K. H., Pluchino, S.: The influence of block of catechol-O-methyl transferase on the sensitivity of isolated organs to catecholamines. Naunyn-Schmiedebergs Arch. Pharmak. 271, 59–92 (1971).
Trendelenburg, U., Graefe, K. H., Eckert, E.: The prejunctional effect of cocaine on the isolated nictitating membrane of the cat. Naunyn-Schmiedebergs Arch. Pharmacol. 275, 69–82 (1972).
Van Orden, L. S. III, Bensch, K. G., Langer, S. Z., Trendelenburg, U.: Histochemical and fine structural aspects of the onset of denervation supersensitivity in the nictitating membrane of the spinal cat. J. Pharmacol. exp. Ther. 157, 274–283 (1967).
Wilkinson, G. N.: Statistical estimations in enzyme kinetics. Biochem. J. 80, 324–332 (1961).
Winne, D.: Zur Auswertung von Versuchsergebnissen: Die Prüfung, ob Kurven sich in ihrem Verlauf unterscheiden. Naunyn-Schmiedebergs Arch. exp. Path. Pharmak. 250, 383–396 (1965).
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Graefe, K.H., Eckert, E. On the stereoselectivity of the neuronal uptake in the cat's nictitating membrane. Naunyn-Schmiedeberg's Arch. Pharmacol. 275, 45–68 (1972). https://doi.org/10.1007/BF00505067
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DOI: https://doi.org/10.1007/BF00505067