Evidence of an exocytotic-like release of [3H]5-hydroxytryptamine induced by d-fenfluramine in rat hippocampal synaptosomes

Marco Gobbi, Emanuela Frittoli, Angela Uslenghi, Tiziana Mennini

Research output: Contribution to journalArticlepeer-review

Abstract

The monoamine releasing activity of d-fenfluramine was investigated with an in vitro model consisting of synaptosomes preloaded with the 3H-neurotransmitter and extensively washed in a superfusion apparatus before a 3-min exposure to d-fenfluramine. With this model, the drug-induced release is real and is not confused by inhibition of reuptake by the drug. d-Fenfluramine (0.5 μM) induced only [3H]5-hydroxytryptamine ([3H]5-HT) release from hippocampal synaptosomes whereas 10 μM also induced some overflow from hippocampal synaptosomes preloaded with [3H]noradrenaline or from striatal synaptosomes preloaded with [3H]dopamine, although the overflow was much lower than from 5-HTergic synaptosomes. We then focused on the [3H]5-HT release induced by 0.5 μM d-fenfluramine, which was previously shown to be Ca2+ dependent. The same finding was confirmed in the present study with other experimental protocols, indicating the requirement for extracellular Ca2+ ions. By measuring [3H]5-HT uptake into rat hippocampal synaptosomes we confirmed that Ca2+-ions are not required for the function of the 5-HT uptake carrier or for its interaction with d-fenfluramine. d-Fenfluramine-induced [3H]5-HT release was not altered by 1 μM nitrendipine (blocking the L-type Ca2+ channels) but was slightly decreased (20%)_by 0.5 μM ω-conotoxin (blocking the N-type Ca2+ channels). It was also inhibited by 0.5 μM clonidine, interacting with α2-adrenergic heteroreceptors, and by 10 nM tetanus toxin, known to affect the exocytosis of different neurotransmitters including 5-HT. These compounds had very similar effects on the Ca2+-dependent, exocytotic release of [3H]5-HT induced by depolarization, i.e. by 15 mMK+. These data suggest that d-fenfluramine, after its carrier-mediated entry into the synaptosomes, induces an influx of extracellular Ca2+, triggering an exocytotic-like release of 5-HT.

Original languageEnglish
Pages (from-to)9-17
Number of pages9
JournalEuropean Journal of Pharmacology
Volume238
Issue number1
DOIs
Publication statusPublished - Jul 6 1993

Keywords

  • (5-hydroxytryptamine, serotonin)
  • 5-HT
  • Cachannels
  • d-Fenfluramine
  • release
  • Synaptosomes

ASJC Scopus subject areas

  • Cellular and Molecular Neuroscience
  • Pharmacology

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