Novel mechanism of modulation at a ligand-gated ion channel; action of 5-Cl-indole at the 5-HT3A Receptor

Research output: Contribution to journalArticlepeer-review

Authors

  • Andrew D Powell
  • Shannon Larkin
  • Nathanael O'Neill
  • Josephine Palandri
  • Reka Otvos
  • Chris Ulens

External organisations

  • School of Clinical and Experimental Medicine, College of Medical and Dental Sciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, UK.
  • Birmingham City University
  • Department of Molecular and Cellular Neurobiology, VU University Amsterdam, Amsterdam, The Netherlands.
  • Laboratory of Structural Neurobiology, KU Leuven, Leuven, Belgium.
  • Department of Pharmacology and Therapeutics, School of Biomedical Sciences, Faculty of Medicine, Dentistry and Health Sciences, The University of Melbourne, Parkville, Victoria, Australia.

Abstract

Background and purpose:
The 5-HT3 receptor is a prototypical member of the Cys-loop ligand-gated ion channel (LGIC) superfamily and an established therapeutic target. In addition to activation via the orthosteric site, receptor function can be modulated by allosteric ligands. We have investigated the pharmacological action of Cl-indole upon the 5- HT3A receptor and identified that this positive allosteric modulator possesses a novel mechanism of action for LGICs.

Experimental approach:
The impact of Cl-indole upon the 5-HT3 receptor was assessed using single cell electrophysiological recordings and [3 H]granisetron binding with HEK293 cells stably expressing the 5-HT3 receptor.

Key results:
Cl-indole failed to evoke 5-HT3A receptor mediated responses (up to 30 µM) or display affinity for the [3 H]granisetron binding site. However, in the presence of Clindole, termination of 5-HT application revealed tail currents mediated via the 5-HT3A receptor that were independent of the preceding 5-HT concentration but were antagonised by the 5-HT3 receptor antagonist, ondansetron. These tail currents were absent in the 5-HT3AB receptor. Furthermore, the presence of 5-HT revealed a concentration-dependent increase in the affinity of Cl-indole for the orthosteric binding site of the h5-HT3A receptor.

Conclusions and implications:
Cl-indole acts as both an orthosteric agonist and an allosteric modulator but the presence of an orthosteric agonist (e.g. 5-HT) is a prerequisite to reveal both actions. Precedent for ago-allosteric action is available yet the essential additional presence of an orthosteric agonist is now reported for the first time. This widening of the pharmacological mechanisms to modulate LGICs may offer further therapeutic opportunities.

Details

Original languageEnglish
Pages (from-to)3467-3479
Number of pages13
JournalBritish Journal of Pharmacology
Volume173
Issue number24
Early online date1 Nov 2016
Publication statusPublished - Dec 2016

Keywords

  • 5-hydroxytryptamine-3 receptor , 5-chloro-indole, ago-allosteric modulator , serotonin , pentameric ligand-gated ion channel , cryptic orthosteric modulator (COM)