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Studies on the nature of the releasable pool of dopamine in synaptosomes from rat corpus striatum: Depolarization-induced release of 3H-dopamine from superfused synaptosomes labelled under various conditions

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Summary

Synaptosomal preparations, obtained from rat corpus striatum, were superfused with Krebs-Ringer medium. During superfusion the synaptosomes were exposed to various concentrations of 3H-dopamine for a few minutes. Depolarization was effected by superfusion with medium containing elevated K+-concentrations. Both the accumulation and the subsequent K+-induced release of 3H-dopamine were quantified in the same preparation. This method was used to detect the presence of 3H-dopamine in different pools in isolated dopaminergic nerve terminals. The data obtained can be summarized as follows:

  1. 1.

    The percentage of 3H-dopamine released and, hence, the distribution of 3H-dopamine into intrasynaptosomal transmitter pools, was independent of the amount of 3H-dopamine accumulated in the synaptosomes.

  2. 2.

    Upon repeated depolarization, 3H-dopamine release was markedly depressed. Supplying the synaptosomes with 3H-dopamine between two stimulations fully restored the release in response to depolarization. Therefore, the depression of 3H-dopamine release upon repeated depolarization is probably due to the selective depletion of a releasable pool and not to an impairment of stimulus-secretion coupling processes.

  3. 3.

    When synaptosomes were subjected twice to K+-stimulation and exposed to 3H-dopamine either before the first stimulation or between both stimulations, the most recently accumulated 3H-dopamine was released (by the second stimulation) in preference to 3H-dopamine accumulated previously.

  4. 4.

    14C-Dopamine synthesized in the synaptosomes from 14C-tyrosine was not released in preference to 3H-dopamine that entered synaptosomal pools through the high affinity uptake mechanism.

  5. 5.

    Our results strongly support the view that dopamine is stored heterogeneously in the varicosities of nigro-striatal neurones in the rat. The remarkably slow redistribution of stored 3H-dopamine into synaptosomal pools indicates that differences in vesicle localization with respect to the nerve terminal membrane might explain the occurrence of different dopamine pools.

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References

  • Akert, K., Pfenniger, K., Sandri, C., Moor, H.: Freeze etching and cytochemistry of vesicles and membrane complexes in synapses of the central nervous system. In: Structure and function of Synapses (G. D. Pappas and D. G. Purpura, eds.), pp. 67–86. New York: Raven Press 1972

    Google Scholar 

  • Bennett, M. R.: An electrophysiological analysis of the storage and release of noradrenaline at sympathetic nerve terminals. J. Physiol. (Lond.) 229, 515–531 (1973)

    Google Scholar 

  • Besson, M. J., Cheramy, A., Feltz, P., Glowinski, J.: Release of newly-synthesized dopamine from dopamine-containing terminals in the striatum of the rat. Proc. Natl. Acad. Sci. USA 62, 741–748 (1969)

    Google Scholar 

  • Besson, M. J., Cheramy, A., Gauchy, C., Glowinski, J.: In vivo continuous estimation of 3H-DA synthesis and release in the cat caudate nucleus: Effects of α-MPT and of transection of the nigro-striatal pathway. Naunyn-Schmiedeberg's Arch. Pharmacol. 278, 101–105 (1973)

    Google Scholar 

  • Costa, E., Carenzi, A., Cheney, D., Guidotti, A., Racagni, G., Zivkovic, B.: Compartmentation of striatal dopamine: problems in assessing the dynamics of functional and storage pools of transmitters. In: Metabolic compartmentation and neurotransmission (S. Berl, D. D. Clarke and D. Schneider, eds.) pp. 167–186. New York, London: Plenum Press 1975

    Google Scholar 

  • Farnebo, L. O., Hamberger, B., Jonsson, G.: Release of 3H-noradrenaline and 3H-dopamine from field stimulated cerebral cortex slices. Effect of tyrosine hydroxylase and dopamine-β-hydroxylase inhibition. J. Neurochem. 18, 2491–2500 (1971)

    Google Scholar 

  • Glowinski, J.: Properties and functions of intraneuronal monoamine compartments in central aminergic neurons. In: Handbook of psychopharmacology (L. L. Iversen, S. D. Iversen and S. H. Snyder, eds.), Vol. 3, pp. 139–167. New York, London: Plenum Press 1975

    Google Scholar 

  • Gray, E. G.: Problems of understanding the substructure of synapses. Progr. Brain Res. 45, 207–234 (1976)

    Google Scholar 

  • Gugten, J. van der, Palkovits, M., Wijnen, H. L. J. M., Versteeg, D. H. G.: Regional distribution of adrenaline in rat brain. Brain Res. 107, 171–175 (1976)

    Google Scholar 

  • Hedqvist, P., Stjärne, L.: The relative role of recapture and of de novo synthesis for the maintenance of neurotransmitter homeostasis in noradrenergic nerves. Acta Physiol. Scand. 76, 270–283 (1969)

    Google Scholar 

  • Heikkila, R. E., Orlanski, H., Cohen, G.: Studies on the distinction between uptake inhibition and release of 3H-dopamine in rat brain tissue slices. Biochem. Pharmacol. 24, 847–852 (1975)

    Google Scholar 

  • Heuser, J. E., Reese, T. S.: Evidence for recycling of synaptic vesicle membrane during transmitter release at the frog neuromuscular junction. J. Cell Biol. 57, 315–344 (1973)

    Google Scholar 

  • Hughes, J.: Differential labelling of intraneuronal noradrenaline stores with different concentrations of (−)3H-noradrenaline. Br. J. Pharmacol. 47, 428–430 (1973)

    Google Scholar 

  • Javoy, F., Glowinski, J.: Dynamic characteristics of the “functional compartment” of dopamine in dopaminergic terminals of the rat striatum. J. Neurochem. 18, 1305–1311 (1971)

    Google Scholar 

  • Kehr, W.: A method for the isolation and determination of 3-methoxy-tyramine in brain tissue. Naunyn-Schmiedeberg's Arch. Pharmacol. 284, 149–158 (1974)

    Google Scholar 

  • Kopin, I. J., Breese, G. R., Krauss, K. R., Weise, V. K.: Selective release of newly synthesized norepinephrine from the cat spleen during sympathetic nerve stimulation. J. Pharmacol. Exp. Ther. 161, 271–278 (1968)

    Google Scholar 

  • Langen, C. D. J. de, Mulder, A. H.: Compartmental analysis of the accumulation of 3H-dopamine in synaptosomes from rat corpus striatum. Naunyn-Schmiedeberg's Arch. Pharmacol. 308, 31–39 (1979)

    Google Scholar 

  • Levy, W. B., Haycock, J. W., Cotman, C. W.: Stimulation-dependent depression of readily releasable neurotransmitter pools in brain. Brain Res. 115, 243–256 (1976)

    Google Scholar 

  • Lowry, O. H., Rosebrough, N. J., Lewis Farr, A., Randall, A. J.: Protein measurements with the folin phenol reagent. J. Biol. Chem. 193, 265–267 (1951)

    Google Scholar 

  • McKenzie, G. M., Szerb, J. C.: The effect of dihydroxyphenylalanine, pheniprazine and dextro-amphetamine on the in vivo release of dopamine from the caudate nucleus. J. Pharmacol. Exp. Ther. 162, 302–308 (1968)

    Google Scholar 

  • Mulder, A. H., Van den Berg, W. B., Stoof, J. C.: Calcium-dependent release of radiolabeled catecholamines and serotonin from rat brain synaptosomes in a superfusion system. Brain Res. 99, 419–424 (1975)

    Google Scholar 

  • Thierry, A. M., Blanc, G., Glowinski, J.: Further evidence for the heterogeneous storage of noradrenaline in central noradrenergic terminals. Naunyn-Schmiedeberg's Arch. Pharmacol. 279, 255–266 (1973)

    Google Scholar 

  • Trendelenburg, U.: Classification of sympathomimetic amines. In: Handbook of experimental pharmacology, Vol. 33 (H. Blaschko and E. Muscholl, eds.), pp. 336–362. Berlin, Heidelberg, New York: Springer 1972

    Google Scholar 

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Authors and Affiliations

  1. Department of Pharmacology, Free University, Medical Faculty, Van der Boechorststraat 7, 1081 BT, Amsterdam, The Netherlands

    Cees D. J. de Langen & Arie H. Mulder

  2. Department of Neurology, Free University, Medical Faculty, Van der Boechorststraat 7, 1081 BT, Amsterdam, The Netherlands

    Johannes C. Stoof

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  1. Cees D. J. de Langen
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  2. Johannes C. Stoof
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  3. Arie H. Mulder
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de Langen, C.D.J., Stoof, J.C. & Mulder, A.H. Studies on the nature of the releasable pool of dopamine in synaptosomes from rat corpus striatum: Depolarization-induced release of 3H-dopamine from superfused synaptosomes labelled under various conditions. Naunyn-Schmiedeberg's Arch. Pharmacol. 308, 41–49 (1979). https://doi.org/10.1007/BF00499717

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  • DOI: https://doi.org/10.1007/BF00499717

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