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PMID: 7620711 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

An electrophysiological investigation of the properties of a murine recombinant 5-HT3 receptor stably expressed in HEK 293 cells.

British journal of pharmacology ·Vol. 114 ·No. 6 ·1995-03-00 ·Pages 1211-21

Gill CH, Peters JA, Lambert JJ

Abstract

1. The pharmacological and biophysical properties of a recombinant 5-HT3 receptor have been studied by use of patch-clamp techniques applied to HEK 293 cells stably transfected with the murine 5-HT3 R-A cDNA. 2. At a holding potential of -60 mV, 77% of cells investigated responded to ionophoretically applied 5-HT with an inward current. Such currents were unaffected by methysergide (1 microM), or ketanserin (1 microM), but were antagonized in a concentration-dependent and reversible manner by the selective 5-HT3 receptor antagonist, ondansetron (IC50 = 440 pM) and the non-selective antagonists (+)-tubocurarine (IC50 = 1.8 nM) and metoclopramide (IC50 50 nM). 3. The 5-HT-induced current reversed in sign (E5-HT) at approximately -2mV and exhibited inward rectification. The influence of extra- and intracellular ion substitutions upon E5-HT indicates the 5-HT-evoked current to be mainly mediated by a mixed monovalent cation conductance. 4. Calcium and magnesium (0.1-10 nM) produced a concentration-dependent, voltage-independent, inhibition of the 5-HT-induced response. Zinc (0.3-300 microM) exerted a biphasic effect with low concentrations enhancing, and high concentrations depressing, the 5-HT-evoked current. 5. Fluctuation analysis of inward currents evoked by a low (1 microM) concentration of 5-HT suggests the current to be mediated by the opening of channels with a conductance of 420 fS. 6. The pharmacological and biophysical properties of the 5-HT3 R-A are similar to those previously described for 5-HT3 receptors native to murine neuroblastoma cell lines, with the exception that the function of the recombinant receptor was enhanced by low concentrations of zinc. This observation suggests that the properties of the native receptor are not completely represented by the 5-HT3 R-A subunit alone.

MeSH Terms
Animals Biophysical Phenomena Biophysics Calcium/pharmacology Cell Line Evoked Potentials/drug effects Magnesium/pharmacology Mice Patch-Clamp Techniques Receptors, Serotonin/physiology Recombinant Proteins/metabolism Transfection Zinc/pharmacology
Chemicals
Receptors, Serotonin Recombinant Proteins Magnesium Zinc Calcium
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Gill C H
Department of Pharmacology and Clinical Pharmacology, Ninewells Hospital and Medical School, University of Dundee.
Peters J A
Lambert J J
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Article Info
Journal
British journal of pharmacology
Abbr.
Br J Pharmacol
ISSN
0007-1188
DOI
10.1111/j.1476-5381.1995.tb13335.x
Published
1995-03-00
Pages
1211-21
Language
English
Region
England
NLM ID
7502536
PMCID
PMC1510359
Subset
IM
Grants
Wellcome Trust · United Kingdom
External Links
PubMed source DOI: 10.1111/j.1476-5381.1995.tb13335.x PMC Full Text View journal details
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