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

Catherine H. Gill, John A. Peters, Jeremy J. Lambert

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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 μm), or ketanserin (1 μm), 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 – 2 mV 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 μm) 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 μm) 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. 1995 British Pharmacological Society

Original languageEnglish
Pages (from-to)1211-1221
Number of pages11
JournalBritish Journal of Pharmacology
Volume114
Issue number6
DOIs
Publication statusPublished - Mar 1995

Fingerprint

HEK293 Cells
Inhibitory Concentration 50
Zinc
Pharmacology
Methysergide
Ondansetron
Tubocurarine
Monovalent Cations
Ketanserin
Metoclopramide
Neuroblastoma
Magnesium
Complementary DNA
Ions
Calcium
Cell Line

Keywords

  • 5‐HT receptor
  • 5‐HT receptor antagonists
  • 5‐HT receptor‐evoked currents
  • 5‐HT single channels
  • recombinant 5‐HT receptor

Cite this

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title = "An electrophysiological investigation of the properties of a murine recombinant 5‐HT3 receptor stably expressed in HEK 293 cells",
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 μm), or ketanserin (1 μm), 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 – 2 mV 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 μm) 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 μm) 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. 1995 British Pharmacological Society",
keywords = "5‐HT receptor, 5‐HT receptor antagonists, 5‐HT receptor‐evoked currents, 5‐HT single channels, recombinant 5‐HT receptor",
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N2 - 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 μm), or ketanserin (1 μm), 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 – 2 mV 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 μm) 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 μm) 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. 1995 British Pharmacological Society

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