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. 1970 Mar;206(3):563–590. doi: 10.1113/jphysiol.1970.sp009031

The extraneuronal uptake and localization of noradrenaline in the cat spleen and the effect on this of some drugs, of cold and of denervation

J S Gillespie, D N H Hamilton, R Jeanette A Hosie
PMCID: PMC1348666  PMID: 5498506

Abstract

1. Extraneuronal uptake of noradrenaline (NA) was examined in the cat spleen first by perfusing with NA for 10 min, followed by a 2-min wash to clear the extracellular fluid, then measuring the amount retained, its subcellular distribution and the tissue components involved as revealed by the development of the characteristic fluorescence. Secondly, thin spleen slices were exposed to NA in vitro and the development of fluorescence in various structures, particularly arterial smooth muscle, measured.

2. The cat spleen accumulated large quantities of NA and this, like the development of fluorescence, was concentration-dependent. After particle separation most of the retained amine appeared in the high-speed supernatant, with a lesser amount in the coarse granule fraction. There was little amine in either the mitochondrial or microsomal fraction. The microsomal fraction from unperfused spleens was rich in NA, presumably from storage granules from the adrenergic nerves. On an intermittent sucrose density gradient the NA-rich particles sedimented between 1·0 and 1·5 M sucrose, corresponding to the recently described dense granules from bovine splenic nerves.

3. Fluorescence histochemistry revealed several tissues accumulating NA. At an NA concentration of 10-5 g/ml., arterial smooth muscle and endothelium showed intracellular fluorescence; at 10-4 g/ml., collagen, the perimeter of the smooth muscle cells of the capsule-trabecula-vein system and the reticular cells forming the framework of the spleen developed fluorescence. In the reticular cells the fluorescence was intracellular. The fluorescence pattern on the perimeter of non-arterial smooth muscle corresponded to the pattern of basement membrane as shown by PAS staining. The red pulp, lymphoid tissue and the phagocytic cells of the ellipsoids did not fluoresce.

4. Cooling the tissue to 15° C or less, phenoxybenzamine in a concentration of 5 × 10-5 g/ml. or normetanephrine in a concentration of 10-4 g/ml. prevented both uptake and loss of NA in arterial smooth muscle but had no effect on collagen.

5. Chronic post-ganglionic denervation or reserpine had no effect on the development of fluorescence in any extraneuronal tissue.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. ANDEN N. E., CARLSSON A., WALDECK B. RESERPINE-RESISTENT UPTAKE MECHANISMS OF NORADRENALINE IN TISSUES. Life Sci. 1963 Dec;12:889–894. doi: 10.1016/0024-3205(63)90056-0. [DOI] [PubMed] [Google Scholar]
  2. Avakian O. V., Gillespie J. S. Uptake of noradrenaline by adrenergic nerves, smooth muscle and connective tissue in isolated perfused arteries and its correlation with the vasoconstrictor response. Br J Pharmacol Chemother. 1968 Jan;32(1):168–184. doi: 10.1111/j.1476-5381.1968.tb00441.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brodie B. B., Costa E., Groppetti A., Matsumoto C. Interaction between desipramine, tyramine, and amphetamine at adrenergic neurones. Br J Pharmacol. 1968 Nov;34(3):648–658. doi: 10.1111/j.1476-5381.1968.tb08494.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Ehinger B., Sporrong B. Neuronal and extraneuronal localization of nonadrenaline in the rat heart after perfusion at high concentration. Experientia. 1968 Mar 15;24(3):265–266. doi: 10.1007/BF02152811. [DOI] [PubMed] [Google Scholar]
  5. Eisenfeld A. J., Axelrod J., Krakoff L. Inhibition of the extraneuronal accumulation and metabolism of norepinephrine by adrenergic blocking agents. J Pharmacol Exp Ther. 1967 Apr;156(1):107–113. [PubMed] [Google Scholar]
  6. Eisenfeld A. J., Landsberg L., Axelrod J. Effect of drugs on the accumulation and metabolism of extraneuronal norepinephrine in the rat heart. J Pharmacol Exp Ther. 1967 Dec;158(3):378–385. [PubMed] [Google Scholar]
  7. FISCHER J. E., KOPIN I. J., AXELROD J. EVIDENCE FOR EXTRANEURONAL BINDING OF NOREPINEPHRINE. J Pharmacol Exp Ther. 1965 Feb;147:181–185. [PubMed] [Google Scholar]
  8. Foo J. W., Jowett A., Stafford A. The effects of some beta-adrenoreceptor blocking drugs on the uptake and release of noradrenaline by the heart. Br J Pharmacol. 1968 Sep;34(1):141–147. doi: 10.1111/j.1476-5381.1968.tb07957.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. GILLESPIE J. S., KIRPEKAR S. M. THE INACTIVATION OF INFUSED NORADRENALINE BY THE CAT SPLEEN. J Physiol. 1965 Jan;176:205–227. doi: 10.1113/jphysiol.1965.sp007545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. GILLIS C. N. CHARACTERISTICS OF NOREPINEPHRINE RETENTION BY A SUBCELLULAR FRACTION OF RABBIT HEART. J Pharmacol Exp Ther. 1964 Oct;146:54–60. [PubMed] [Google Scholar]
  11. Gillespie J. S., Hamilton D. N. Binding of noradrenaline to smooth muscle cells in the spleen. Nature. 1966 Oct 29;212(5061):524–525. doi: 10.1038/212524a0. [DOI] [PubMed] [Google Scholar]
  12. Gillespie J. S., Kirpekar S. M. The histological localization of noradrenaline in the cat spleen. J Physiol. 1966 Nov;187(1):69–79. doi: 10.1113/jphysiol.1966.sp008076. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. HERTTING G., AXELROD J. Fate of tritiated noradrenaline at the sympathetic nerve-endings. Nature. 1961 Oct 14;192:172–173. doi: 10.1038/192172a0. [DOI] [PubMed] [Google Scholar]
  14. Hamberger B., Malmfors T. Uptake and release of alpha-methyl-noradrenaline in vitro after reserpine pretreatment. A histochemical study. Acta Physiol Scand. 1967 Jul-Aug;70(3):412–418. doi: 10.1111/j.1748-1716.1967.tb03639.x. [DOI] [PubMed] [Google Scholar]
  15. Hamberger B., Norberg K. A., Olson L. Extraneuronal binding of catecholamines and 3,4-dihydroxyphenylalanine (dopa) in salivary glands. Acta Physiol Scand. 1967 Jan-Feb;69(1):1–12. doi: 10.1111/j.1748-1716.1967.tb03485.x. [DOI] [PubMed] [Google Scholar]
  16. Hedqvist P., Oliverio A., Stjärne L. Inhibition by phenoxybenzamine of the noradrenaline releasing effect of tyramine. Acta Physiol Scand. 1968 Apr;72(4):385–391. doi: 10.1111/j.1748-1716.1968.tb03862.x. [DOI] [PubMed] [Google Scholar]
  17. Horst W. D., Kopin I. J., Ramey E. R. Influence of sodium and calcium on norepinephrine uptake by isolated perfused rat hearts. Am J Physiol. 1968 Oct;215(4):817–822. doi: 10.1152/ajplegacy.1968.215.4.817. [DOI] [PubMed] [Google Scholar]
  18. Häggendal J., Hamberger B. Quantitative in vitro studies on noradrenaline uptake and its inhibition by amphetamine, desipramine and chlorpromazine. Acta Physiol Scand. 1967 Jul-Aug;70(3):277–280. doi: 10.1111/j.1748-1716.1967.tb03626.x. [DOI] [PubMed] [Google Scholar]
  19. IVERSEN L. L. THE UPTAKE OF ADRENALINE BY THE RAT ISOLATED HEART. Br J Pharmacol Chemother. 1965 Apr;24:387–394. doi: 10.1111/j.1476-5381.1965.tb01726.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. IVERSEN L. L. THE UPTAKE OF NORADRENALINE BY THE ISOLATED PERFUSED RAT HEART. Br J Pharmacol Chemother. 1963 Dec;21:523–537. doi: 10.1111/j.1476-5381.1963.tb02020.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Iversen L. L., Glowinski J., Axelrod J. The uptake and storage of H3-norepinephrine in the reserpine-pretreated rat heart. J Pharmacol Exp Ther. 1965 Nov;150(2):173–183. [PubMed] [Google Scholar]
  22. KIRSHNER N. Uptake of catecholamines by a particulate fraction of the adrenal medulla. J Biol Chem. 1962 Jul;237:2311–2317. [PubMed] [Google Scholar]
  23. KOPIN I. J., GORDON E. K. Metabolism of administered and drug-released norepinephrine-7-H3 in the rat. J Pharmacol Exp Ther. 1963 May;140:207–216. [PubMed] [Google Scholar]
  24. Kalsner S., Nickerson M. Effects of a haloalkylamine on responses to and disposition of sympathomimetic amines. Br J Pharmacol. 1969 Mar;35(3):440–455. doi: 10.1111/j.1476-5381.1969.tb08285.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Kalsner S., Nickerson M. Effects of reserpine on the disposition of sympathomimetic amines in vascular tissue. Br J Pharmacol. 1969 Mar;35(3):394–405. doi: 10.1111/j.1476-5381.1969.tb08281.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. LINDMAR R., MUSCHOLL E. DIE WIRKUNG VON PHARMAKA AUF DIE ELIMINATION VON NORADRENALIN AUS DER PERFUSIONSFLUESSIGKEIT UND DIE NORADRENALINEUFNAHME IN DAS ISOLIERTE HERZ. Naunyn Schmiedebergs Arch Exp Pathol Pharmakol. 1964 Jul 28;247:469–492. doi: 10.1007/BF00329896. [DOI] [PubMed] [Google Scholar]
  27. POTTER L. T., AXELROD J. SUBCELLULAR LOCALIZATION OF CATECHOLAMINES IN TISSUES OF THE RAT. J Pharmacol Exp Ther. 1963 Dec;142:291–298. [PubMed] [Google Scholar]
  28. POTTER L. T., COOPER T., WILLMAN V. L., WOLFE D. E. SYNTHESIS, BINDING, RELEASE, AND METABOLISM OF NOREPINEPHRINE IN NORMAL AND TRANSPLANTED DOG HEARTS. Circ Res. 1965 May;16:468–481. doi: 10.1161/01.res.16.5.468. [DOI] [PubMed] [Google Scholar]
  29. Roth R. H., Stjärne L., Bloom F. E., Giarman N. J. Light and heavy norepinephrine storage particles in the rat heart and in bovine splenic nerve. J Pharmacol Exp Ther. 1968 Aug;162(2):203–212. [PubMed] [Google Scholar]
  30. SCHUEMANN H. J., WEIGMANN E. [On the point of attack of the indirect action of sympathomimetic amines]. Naunyn Schmiedebergs Arch Exp Pathol Pharmakol. 1960;240:275–284. [PubMed] [Google Scholar]
  31. SNYDER S. H., MICHAELSON I. A., MUSACCHIO J. PURIFICATION OF NOREPINEPHRINE STORAGE GRANULES FROM RAT HEART. Life Sci. 1964 Sep;3:965–970. doi: 10.1016/0024-3205(64)90106-7. [DOI] [PubMed] [Google Scholar]
  32. STROMBLAD B. C., NICKERSON M. Accumulation of epinephrine and norepinephrine by some rat tissues. J Pharmacol Exp Ther. 1961 Nov;134:154–159. [PubMed] [Google Scholar]
  33. Sjöqvist F., Taylor P. W., Jr, Titus E. The effect of immunosympathectomy on the retention and metabolism of noradrenaline. Acta Physiol Scand. 1967 Jan-Feb;69(1):13–22. doi: 10.1111/j.1748-1716.1967.tb03486.x. [DOI] [PubMed] [Google Scholar]
  34. WEGMANN A., KAKO K. Particle-bound and free catecholamines in dog hearts and the uptake of injected norepinephrine. Nature. 1961 Dec 9;192:978–978. doi: 10.1038/192978a0. [DOI] [PubMed] [Google Scholar]
  35. WOLFE D. E., POTTER L. T., RICHARDSON K. C., AXELROD J. Localizing tritiated norepinephrine in sympathetic axons by electron microscopic autoradiography. Science. 1962 Oct 19;138(3538):440–442. doi: 10.1126/science.138.3538.440. [DOI] [PubMed] [Google Scholar]
  36. von EULER U., LISHAJKO F. Improved technique for the fluorimetric estimation of catecholamines. Acta Physiol Scand. 1961 Apr;51:348–355. doi: 10.1111/j.1748-1716.1961.tb02128.x. [DOI] [PubMed] [Google Scholar]
  37. von Euler U. S., Lishajko F. Inhibitory action of adrenergic blocking agents on reuptake and net uptake of noradrenaline in nerve granules. Acta Physiol Scand. 1968 Nov;74(3):501–506. doi: 10.1111/j.1748-1716.1968.tb04258.x. [DOI] [PubMed] [Google Scholar]
  38. von Euler U. S. Release and uptake of noradrenaline in adrenergic nerve granules. Acta Physiol Scand. 1966 Jul-Aug;67(3):430–440. doi: 10.1111/j.1748-1716.1966.tb03330.x. [DOI] [PubMed] [Google Scholar]

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