Utility of organotypic raphe slice cultures to investigate the effects of sustained exposure to selective 5-HT reuptake inhibitors on 5-HT release

Br J Pharmacol. 2010 Dec;161(7):1527-41. doi: 10.1111/j.1476-5381.2010.00978.x.

Abstract

Background and purpose: Selective 5-hydroxytryptamine (5-HT, serotonin) reuptake inhibitors (SSRIs) are widely used antidepressants and their therapeutic effect requires several weeks of drug administration. The delayed onset of SSRI efficacy is due to the slow neuroadaptive changes of the 5-hydroxytryptaminergic (5-HTergic) system. In this study, we examined the acute and chronic effects of SSRIs on the 5-HTergic system using rat raphe slice cultures.

Experimental approach: For organotypic raphe slice cultures, mesencephalic coronal sections containing dorsal and median raphe nuclei were prepared from neonatal Wistar rats and cultured for 14-16 days.

Key results: Acute treatment with citalopram, paroxetine or fluoxetine (0.1-10 µM) in the slice cultures slightly increased extracellular 5-HT levels, while sustained exposure for 4 days augmented the elevation of 5-HT level in a time-dependent manner. Sustained exposure to citalopram had no effect on tissue contents of 5-HT and its metabolite, expression of tryptophan hydroxylase or the membrane expression of 5-HT transporters. The augmented 5-HT release was attenuated by Ca(2+) -free incubation medium or treatment with tetrodotoxin. Experiments with 5-HT(1A/B) receptor agonists and antagonists revealed that desensitization of 5-HT(1) autoreceptors was not involved in the augmentation of 5-HT release. Finally, co-treatment with an α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)/kainate, but not an N-methyl-d-aspartate, receptor antagonist, suppressed this augmentation.

Conclusion and implications: These results suggest that sustained exposure to SSRIs induces the augmentation of exocytotic 5-HT release, which is caused, at least in part, by the activation of AMPA/kainate receptors in the raphe slice cultures.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Action Potentials / drug effects
  • Animals
  • Antidepressive Agents / pharmacology*
  • Autoreceptors / metabolism
  • Brain / drug effects*
  • Brain / metabolism
  • Calcium / metabolism
  • Citalopram / pharmacology
  • Extracellular Space / metabolism
  • Fluoxetine / pharmacology
  • Hydroxyindoleacetic Acid / metabolism
  • Organ Culture Techniques
  • Paroxetine / pharmacology
  • Raphe Nuclei / drug effects*
  • Raphe Nuclei / metabolism
  • Rats
  • Rats, Wistar
  • Selective Serotonin Reuptake Inhibitors / pharmacology*
  • Serotonin / metabolism*
  • Serotonin Plasma Membrane Transport Proteins / metabolism
  • Time Factors
  • Tryptophan Hydroxylase / metabolism

Substances

  • Antidepressive Agents
  • Autoreceptors
  • Serotonin Plasma Membrane Transport Proteins
  • Serotonin Uptake Inhibitors
  • Fluoxetine
  • Citalopram
  • Serotonin
  • Paroxetine
  • Hydroxyindoleacetic Acid
  • Tryptophan Hydroxylase
  • Calcium