Skip to main content
Log in

Critical assessment of noradrenaline uptake in synaptosomal preparations

  • Published:
Naunyn-Schmiedeberg's Archives of Pharmacology Aims and scope Submit manuscript

Summary

Synaptosomes and other subcellular organelles were prepared from rat brain using a vertical rotor. The preparation time was reduced by up to 60% compared to conventional techniques. Uptake of [3H]-(−)-noradrenaline into subcellular fractions was characterised. The characteristics of this uptake were dependent on the subcellular composition and anatomical origin. Various methods of correction for energy independent processes were compared, but only sodium ion removal from the medium selectively inhibited the energy dependent uptake mechanism. Kinetic analysis of data revealed that high and low affinity uptake systems existed. The relative contributions of these systems were dependent on the fraction under analysis. Noradrenaline uptake was not exclusively localised in noradrenergic terminals. Selective inhibitors of the noradrenaline uptake process (tricyclic antidepressants) inhibited energy dependent uptake completely only in purified synaptosomes prepared from cortex. In whole brain synaptosomal fractions, noradrenaline was partially accumulated into dopaminergic neurones; this uptake process was not inhibited by tricyclic antidepressants.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

Abbreviations

(−)-NA:

(−)-noradrenaline

LiCl:

lithium chloride

NaCl:

sodium chloride

References

  • Bogdanski DF (1976) Mechanisms of transport for the uptake and release of biogenic amines in nerve endings. Adv Exp Med Biol 69:291–305

    Google Scholar 

  • Cleaver GJ, Kotas AM, Prince AK, Reynolds R, Wong PT-H (1980) The uptake of choline into synaptosomes; non-Michaelis-Menten kinetics demonstrated by a grouped least squares analysis. Br J Pharmacol 69:337P-339P

    Google Scholar 

  • Cotman CW, Haycock JW, White WF (1976) Stimulus-secretion coupling process in brain: analysis of noradrenaline and gamma-aminobutyric acid release. J Physiol (Lond) 254:475–505

    Google Scholar 

  • De Belleroche JS, Bradford HF (1973) The synaptosome: an isolated, working, neuronal compartment. In: Kerbut GA, Phillis JW (eds) Progress in neurobiology. Pergamon Press, Oxford, pp 275–298

    Google Scholar 

  • De Lean A, Munson PJ, Rodbard D (1978) Simultaneous analysis of families of sigmoidal curves: application to bioassay, radioligand assay and physiological dose-response curves. Am J Physiol 235:E97-E102

    Google Scholar 

  • Eckhardt JB, Maxwell RA (1980) Norepinephrine uptake in cortical and hypothalamic synaptosomes; differences in response to tricyclic anti-depressants. Fed Proc 64th Annual Meeting, part II

  • Gilbert JC, Wyllie MG (1976) Effects of anticonvulsant and convulsant drugs on ATPase activities of synaptosomes and their components. Br J Pharmacol 56:49–57

    Google Scholar 

  • Gilbert JC, Wyllie MG (1981) The relationship between nerve terminal adenosine triphosphatases and neurotransmitter release: as determined by the use of antidepressant and other CNS active drugs. Br J Pharmacol 69:215–225

    Google Scholar 

  • Glowinski J, Iversen LL (1966) Regional studies of catecholamine in the rat brain. I. The disposition of [3H]norepinephrine, [3H]dopamine and [3H]DOPA in various regions of the brain. J Neurochem 13: 655–671

    Google Scholar 

  • Graefe KH (1976) Methodology of catecholamine transport studies: definition of terms. In: Paton DM (ed) Mechanism of neuronal and extraneuronal transport of catecholamines. Raven Press, New York, pp 7–35

    Google Scholar 

  • Gray EG, Whittaker VP (1962) The isolation of nerve endings from brain: an electronmicroscopic study of cell fragments derived by homogenisation and centrifugation. J Anat 96:79–88

    Google Scholar 

  • Hokin LE (1974) Purification and properties of the (sodium + potassium) activated adenosinetriphosphatase and reconstitution of sodium transport. Ann NY Acad Sci 242:12–23

    Google Scholar 

  • Iversen LL (1971) Role of transmitter uptake in synaptic neurotransmission. Br J Pharmacol 41:571–591

    Google Scholar 

  • Iversen LL (1973) Neuronal and extraneuronal catecholamine uptake mechanisms. In: Usdin E, Snyder SH (eds) Frontiers in catecholamine research. Pergamon Press, Oxford, pp 403–408

    Google Scholar 

  • Iversen LL (1975) Uptake processes for biogenic amines. In: Iversen LL, Iversen SD, Snyder SH (eds) Handbook of psychopharmacology, vol. 3: Biochemistry of biogenic amines. Plenum Press, New York, pp 381–442

    Google Scholar 

  • Jones DG (1975) Synapses and synaptosomes, morphological aspects. Chapman and Hall, London

    Google Scholar 

  • Koe BK (1976) Molecular geometry of inhibitors of the uptake of catecholamines and serotonin in synaptosomal preparations of rat brain. J Pharmacol Exp Ther 199:649–661

    Google Scholar 

  • Kolber AR, Bagnell CR, Kringman MR, Hayward J, Morell P (1979) Transport of sugars into microvessels isolated from the rat brain: a model for the blood brain barrier. J Neurochem 33:419–431

    Google Scholar 

  • Kuriyama K, Weinstein H, Roberts E (1969) Uptake of γ-aminobutyric acid by mitochondrial and synaptosomal fractions from mouse brain. Brain Res 16:479–492

    Google Scholar 

  • Levi G, Raiteri M (1973) GABA and glutamate uptake by subcellular fractions enriched in synaptosomes: critical evaluation of some methological aspects. Brain Res 57:165–186

    Google Scholar 

  • Levi G, Raiteri M (1976) Synaptosomal transport processes, Int Rev Neurobiol 19:51–74

    Google Scholar 

  • Minchin MC (1979) Uptake of [14C]-nipecotic acid into rat dorsal root ganglia. J Neurochem 32:1519–1524

    Google Scholar 

  • Neal JL (1972) Analysis of Michaelis kinetics for two independent saturable membrane transport functions. J Theor Biol 35:113–118

    Google Scholar 

  • O'Fallon JV, Brosemar RW, Harding JW (1981) The Na+, K+-ATPase: a plausible trigger for voltage-independent release of cytoplasmic neurotransmitter. J Neurochem 36:369–378

    Google Scholar 

  • O'Hanlon JF, Campuzano HC, Horvath SM (1970) A fluorimetric assay for subnanogram concentrations of adrenaline and noradrenaline in plasma. Anal Biochem 34:568–581

    Google Scholar 

  • Paton DM (1976) Characteristics of uptake of noradrenaline by adrenergic neurones. In: Paton DM (ed) The mechanisms of neuronal and extraneuronal transport of catecholamines. Raven Press, New York, pp 49–65

    Google Scholar 

  • Post C, Lewis DH (1979) Displacement of nortriptyline and uptake of 14C-lidocaine in the lung after administration of 14C-lidocaine to nortriptyline intoxicated pigs. Acta Pharmacol Toxicol 45:218–224

    Google Scholar 

  • Romer J, Bickel MH (1979) Interaction of chlorpromazine and imipramine with artificial membranes investigated by equilibrium dialysis, dual wavelength photometry and fluorimetry. Biochem Pharmacol 28:799–805

    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 

  • Shaskan EG, Snyder SH (1975) Kinetics of serotonin accumulation into slices from rat brain: relationship to catecholamine uptake. J Pharmacol Exp Ther 175:404–418

    Google Scholar 

  • Simon JR, Atweh S, Kuhar MJ (1976) Sodium-dependent high affinity choline uptake: a regulatory step in the synthesis of acetylcholine. J Neurochem 26:909–922

    Google Scholar 

  • Vosmer G, De Met E, Halaris AE (1980) Action of the antidepressant pridefine (AHR-118) on biogenic amines in the rat brain. Biochem Pharmacol 29:2557–2562

    Google Scholar 

  • Wheeler DD (1978) Some problems inherent in transport studies in synaptosomes. J Neurochem 30:109–120

    Google Scholar 

  • Wheeler DD (1979) A model of high affinity glutamic acid transport by rat cortical synaptosomes-a refinement of the originally proposed model. J Neurochem 33:883–894

    Google Scholar 

  • Wheeler DD (1980) Sodium dependence of GABA transport in rat hypothalamic synaptosomes. J Neurosci Res 5:323–337

    Google Scholar 

  • Wong DT, Bymaster FP, Reid LR (1980) Competitive inhibition of catecholamine uptake in synaptosomes of ratbrain by rigid bicyclooctanes. J Neurochem 34:1453–1458

    Google Scholar 

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

    Google Scholar 

  • Wood MD, Wyllie MG (1981) The rapid preparation of synaptosomes using a vertical rotor. J Neurochem 37:795–797

    Google Scholar 

  • Wyllie MG, Gilbert JC (1980) Exocytotic release of noradrenaline from synaptosomes. Biochem Pharmacol 29:1302–1303

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wood, M.D., Wyllie, M.G. Critical assessment of noradrenaline uptake in synaptosomal preparations. Naunyn-Schmiedeberg's Arch. Pharmacol. 322, 129–135 (1983). https://doi.org/10.1007/BF00512385

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00512385

Key words

Navigation