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Uptake of 14C-tyramine and release of extravesicular 3H-noradrenaline in isolated perfused rabbit hearts

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Summary

  1. 1)

    In isolated perfused hearts of reserpine (R)-and pargyline (P)-pretreated rabbits (i.e. RP-hearts) initial rates of intracellular uptake of 14C-tyramine and of 3H-amphetamine were measured as described by Graefe et al. (1978). The uptake of 14C-tyramine, but not that of 3H-amphetamine was inhibited by cocaine.

  2. 2)

    Saturation kinetics of the intracellular uptake of 14C-tyramine revealed a non-saturable (diffusional) and a saturable (carrier-mediated) component of uptake.

  3. 3)

    After 30 min of perfusion with 14C-tyramine the accumulated 14C-radioactivity in RP-hearts consisted of unchanged tyramine (about 60%), octopamine (about 30%) and deaminated metabolites (about 10%). In contrast, 14C-octopamine was the main radioactive substance when the perfusion with 14C-tyramine was followed by 100 min of wash-out.

  4. 4)

    IC50-Values of tyramine, amphetamine, amantadine and nomifensine for inhibition of neuronal uptake of 3H-noradrenaline were determined in RPU-hearts (i.e. in RP-hearts whose COMT was inhibited by U-0521).

  5. 5)

    About equieffective concentrations (with respect to inhibition of 3H-noradrenaline uptake) of tyramine, amphetamine, amantadine and noradrenaline (i.e. of substrates of the neuronal amine carrier) caused a pronounced (and comparable) release of 3H-noradrenaline from RPU-hearts, whereas cocaine and nomifensine (i.e. uptake inhibitors) caused only a very small release. Low sodium caused a release comparable to that induced by substrates of the amine carrier.

  6. 6)

    Increasing concentrations of tyramine (0.2–24 μmol/l) caused mobilization of 3H-noradrenaline from a small “bound fraction” and partial mobilization from a large compartment which was characterized by a rate constant for efflux of about 0.014 min−1 (compartment I). The peak-value of the tyramine-induced efflux of 3H-noradrenaline exhibited saturability with increasing concentrations of tyramine. Half-maximal release was observed at a tyramine concentration which corresponded to a) the IC50-value for inhibition of uptake of 3H-noradrenaline and b) the K m of the saturable component of uptake of 14C-tyramine.

  7. 7)

    That part of neuronally accumulated 3H-noradrenaline (mainly in compartment I) which was not further mobilized by high concentrations of tyramine was also hardly mobilized by veratridine (in the absence of Ca2+). However, in the presence of Ca2+, veratridine as well as nicotine induced a release of this “tyramine-resistant” 3H-noradrenaline.

  8. 8)

    It is concluded that in RPU-hearts the distribution of 3H-noradrenaline within the partially “tyramine-resistant” compartment I and within the “bound fraction” might represent 3H-noradrenaline “trapped” within the acid interior of “reserpinized” vesicles and within a small population of intact storage vesicles, respectively. The fast release of 3H-noradrenaline (from RPU-hearts) by tyramine, noradrenaline, amphetamine and amantadine might be caused by facilitation of the outward transport of axoplasmic noradrenaline; the extend of facilitation may be directly connected to the velocity of uptake of these substrates by the amine carrier.

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References

  • Andén NE, Corrodi H, Ettles M, Gustafson E, Persson H (1964) Selective uptake of some catecholamines by the isolated heart and its inhibition by cocaine and phenoxybenzamine. Acta Pharmacol Toxicol 21:247–258

    Google Scholar 

  • Bönisch H (1981) Evidence for a cocaine-sensitive and sodium-dependent uptake of amphetamine. Naunyn-Schmiedeberg's Arch Pharmacol (Suppl) 316:R54

    Google Scholar 

  • Bönisch H, Graefe KH, Keller B (1980) Reserpine-like, tetrodotoxin (TTX)-resistant effect of veratridine in the rat vas deferens. Naunyn-Schmiedeberg's Arch Pharmacol (Suppl) 311:R61

    Google Scholar 

  • Brandao F, Rodrigues-Pereira E, Guilherme Monteiro J, Davidson R (1981) A kinetic study of the release of noradrenaline by tyramine. Naunyn-Schmiedeberg's Arch Pharmacol 318:83–87

    Google Scholar 

  • Carlsson A, Waldeck B (1966) Effects of amphetamine, tyramine, and protryptyline on reserpine-resistant amine-concentrating mechanisms of adrenergic nerves. J Pharm Pharmacol 18:252–253

    Google Scholar 

  • Carlsson A, Fuxe K, Hamberger B, Lindqvist M (1966) Biochemical and histochemical studies on the effect of imipramine-like drugs and (+)amphetamine on central and peripheral catecholamine neurons. Acta Physiol Scand 67:481–492

    Google Scholar 

  • Enna SJ, Shore PA (1974) On the nature of the adrenergic neurone extragranular amine binding site. J Neural Transmiss 35:125–135

    Google Scholar 

  • Evoniuk G, Slotkin TA (1981) pH Dependence of (3H)norepinephrine uptake into catecholamine storage vesicles isolated from rat brain, heart and adrenal medulla. Biochem Pharmacol 30:2872–2873

    Google Scholar 

  • Farnebo LO (1971) Effects of reserpine on release of 3H-noradrenaline, 3H-dopamine, and 3H-metaraminol from field stimulated rat iris. Biochem Pharmacol 20:2715–2726

    Google Scholar 

  • Famebo LO, Fuxe K, Goldstein M, Hamberger B, Ungerstedt U (1971) Dopamine and noradrenaline releasing action of amantadine in the central and peripheral nervous system: A possible mode of action in Parkinson's disease. Eur J Pharmacol 16:27–38

    Google Scholar 

  • Fischer JF, Cho AK (1979) Chemical release of dopamine from striatal homogenates: evidence for an exchange diffusional model. J Pharmacol Exp Ther 208:203–209

    Google Scholar 

  • Giachetti A, Hollenbeck RA (1976) Extra-vesicular binding of noradrenaline and guanethidine in the adrenergic neurones of the rat heart: a proposed site of adrenergic neurone blocking agents. Br J Pharmacol 58:497–504

    Google Scholar 

  • Giles RE, Miller JW (1967) Studies on the potentiation of the inotropic actions of certain catecholamines by U-0521 (3′,4′-dihydroxymethyl propiophenone). J Pharmacol Exp Ther 157:55–61

    Google Scholar 

  • Golko DS, Paton DM (1976) Characteristics of accumulation of ephedrine in rabbit atria. Can J Physiol Pharmacol 54:93–100

    Google Scholar 

  • Graefe KH (1981) The disposition of 3H-(−)noradrenaline in the perfused cat and rabbit heart. Naunyn-Schmiedeberg's Arch Pharmacol 318:71–82

    Google Scholar 

  • Graefe KH, Trendelenburg U (1974) The effect of hydrocortisone on the sensitivity of the isolated nictitating membrane to catecholamines: relationship to extraneuronal uptake and metabolism. Naunyn-Schmiedeberg's Arch Pharmacol 286:7–48

    Google Scholar 

  • Graefe KH, Stefano FJE, Langer SZ (1973) Preferential metabolism of 3H-(−)norepinephrine through the deaminated glycol in the rat vas deferens. Biochem Pharmacol 22:1147–1160

    Google Scholar 

  • Graefe KH, Stefano FJE, Langer SZ (1977) Stereoselectivity in the metabolism of 3H-noradrenaline during uptake into and efflux from the rat vas deferens. Naunyn-Schmiedeberg's Arch Pharmacol 299:225–238

    Google Scholar 

  • Graefe KH, Bönisch H, Keller B (1978) Saturation of the adrenergic neurone uptake system in the perfused rabbit heart. A new method for determination of initial rates of amine uptake. Naunyn-Schmiedeberg's Arch Pharmacol 302:263–273

    Google Scholar 

  • Henseling M, Eckert E, Trendelenburg U (1976) The distribution of 3H-(±)-noradrenaline in rabbit aortic strips after inhibition of the noradrenaline-metabolizing enzymes. Naunyn-Schmiedeberg's Arch Pharmacol 292:205–217

    Google Scholar 

  • Holbach HJ, Lindmar R, Löffelholz K (1977) DMPP and the adrenergic nerve terminal: mechanisms of noradrenaline release from vesicular and extravesicular compartments. Naunyn-Schmiedeberg's Arch Pharmacol 300:131–138

    Google Scholar 

  • Holz RW (1978) Evidence that catecholamine transport into chromaffine vesicles is coupled to vesicle membrane potential. Proc Natl Acad Sci USA 75:5190–5194

    Google Scholar 

  • Johnson RG, Scarpa A (1976) Internal pH of isolated chromaffin vesicles. J Biol Chem 251:2189–2191

    Google Scholar 

  • Lindmar R, Löffelholz K (1974) Neuronal and extraneuronal uptake and efflux of catecholamines in the isolated rabbit heart. Naunyn-Schmiedeberg's Arch Pharmacol 284:63–92

    Google Scholar 

  • Lundberg P, Stitzel R (1967) Uptake of biogenic amines by two different mechanisms present in adrenergic granules. Br J Pharmacol 29:342–349

    Google Scholar 

  • Lundberg P, Waldeck B (1971) On the mechanism of amphetamine induced release of reserpine-resistant 3H-noradrenaline and 3H-methylnoradrenaline. Acta Pharmacol Toxicol 30:339–347

    Google Scholar 

  • Mack F, Bönisch H (1979) Dissociation constants and lipophilicity of catecholamines and related compounds. Naunyn-Schmiedeberg's Arch Pharmacol 310:1–9

    Google Scholar 

  • Mekanontchai R, Trendelenburg U (1979) The neuronal and extra-neuronal distribution of 3H(−)noradrenaline in the perfused rat heart. Naunyn-Schmiedeberg's Arch Pharmacol 308:199–210

    Google Scholar 

  • Paton DM (1973) Mechanism of efflux of noradrenaline from adrenergic nerves in rabbit atria. Br J Pharmacol 49:614–627

    Google Scholar 

  • Paton DM (1976) Characteristics of efflux of noradrenaline from adrenergic neurons. In: Paton DM (ed) The mechanism of neuronal and extraneuronal transport of catecholamines. Raven Press, New York, pp 155–174

    Google Scholar 

  • Pereira ER, Bönisch H (1978) The uptake of tyramine and the release by tyramine of extravesicular noradrenaline in the isolated perfused hearts of reserpine pretreated rabbits. In: Proceedings of the 3rd Meeting on Adrenergic Mechanisms, Porto, pp 70–71

  • Raiteri M, Bertolini A, Angelini F, Levi G (1975) d-Amphetamine as a releaser or reuptake inhibitor of biogenic amines in synaptosomes. Eur J Pharmacol 34:189–195

    Google Scholar 

  • Ross SB, Kelder D (1976) Effect of veratridine on the fluxes of 3H-noradrenaline and 3H-bretylium in the rat vas deferens in vitro. Naunyn-Schmiedeberg's Arch Pharmacol 295:183–189

    Google Scholar 

  • Ross SB, Kelder D (1977) Evoked efflux of 3H-bretylium by sympathomimetic amines from the rat vas deferens in vitro. Acta Pharmacol Toxicol 40:87–96

    Google Scholar 

  • Ross SB, Renyi AL (1966) Uptake of tritiated tyramine and (+)amphetamine by mouse heart slices. J Pharm Pharmacol 18:756–757

    Google Scholar 

  • Ross SB, Renyi AL (1978) Effect of (+)amphetamine on the retention of 3H-catecholamines in slices of normal and reserpinized rat brain and heart. Acta Pharmacol Toxicol 42:328–336

    Google Scholar 

  • Rutledge CO, Vollmer S (1979) Evidence for carrier mediated efflux of noradrenaline displaced by amphetamine. In: Usdin E, Kopin IJ, Barchas J (eds) Catecholamines; Basic and clinical frontiers, vol 1. Pergamon Press, New York, pp 304–306

    Google Scholar 

  • Sammet S, Graefe KH (1979) Kinetic analysis of the interaction between noradrenaline and Na+ in neuronal uptake: kinetic evidence for cotransport. Naunyn-Schmiedeberg's Arch Pharmacol 309:99–107

    Google Scholar 

  • Snedecor GW, Cochran WG (1976) Statistical methods (6th edtion). Iowa State University Press, Ames

    Google Scholar 

  • Steinberg MI, Smith CB (1971) Uptake, retention and metabolism of 3H-tyramine in rat atria. J Pharmacol Exp Ther 176:139–148

    Google Scholar 

  • Thoenen H, Hürlimann A, Haefely W (1968) Mechanism of amphetamine accumulation in the isolated perfused heart of the rat. J Pharm Pharmacol 20:1–11

    Google Scholar 

  • Trendelenburg U (1978) Release induced by phenethylamines. In: Paton DM (ed) The release of catecholamines from adrenergic neurons. Pergamon Press, New York: 333–354

    Google Scholar 

  • Trendelenburg U, Bönisch H, Graefe KH, Henseling M (1980) The rate constants for the efflux of metabolites of catecholamines and phenethylamines. Pharmacol Rev. 31:179–203

    Google Scholar 

  • Van Orden LD, Schaefer JM, Antonaccio MJ, Smith CB (1974) Fine structural identification of a reserpine-resistant norepinephrine store in adrenergic nerve terminals in guinea pig atria. J. Pharmacol Exp Ther 188:668–675

    Google Scholar 

  • Wilkinson GN (1961) Statistical estimation in enzyme kinetics. Biochem. J 80:324–332

    Google Scholar 

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Author notes

  1. E. Rodrigues-Pereira (E.R.-P. obtained a fellowship from the Deutscher Akademischer Austauschdienst (DAAD)

    Present address: Laboratorio de Farmacologia, Faculdade de Medicina, Porto, Portugal

Authors and Affiliations

  1. Institut für Pharmakologie und Toxikologie der Universität Würzburg, Versbacher Strasse 9, D-8700, Würzburg, Federal Republic of Germany

    H. Bönisch & E. Rodrigues-Pereira (E.R.-P. obtained a fellowship from the Deutscher Akademischer Austauschdienst (DAAD)

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  1. H. Bönisch
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  2. E. Rodrigues-Pereira
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Bönisch, H., Rodrigues-Pereira, E. Uptake of 14C-tyramine and release of extravesicular 3H-noradrenaline in isolated perfused rabbit hearts. Naunyn-Schmiedeberg's Arch. Pharmacol. 323, 233–244 (1983). https://doi.org/10.1007/BF00497669

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  • DOI: https://doi.org/10.1007/BF00497669

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