Electrophysiological and pharmacological properties of the human brain type IIA Na+ channel expressed in a stable mammalian cell line

Pflugers Arch. 2001 Jan;441(4):425-33. doi: 10.1007/s004240000448.

Abstract

The human brain voltage-gated Na+ channel type IIA alpha subunit was cloned and stably expressed in Chinese hamster ovary cells and its biophysical and pharmacological properties were studied using whole-cell voltage-clamp. Fast, transient inward currents of up to -8,000 pA were elicited by membrane depolarization of the recombinant cells. Channels activated at -50 mV and reached maximal activation at -10 mV to 0 mV. The reversal potential was 62 +/- 2 mV which is close to the Na+ equilibrium potential. The half-maximal activation and inactivation voltages were -24 +/- 2 mV and -63 +/- 1 mV, respectively. Currents were reversibly blocked by tetrodotoxin with a half-maximal inhibition of 13 nM. The effects of four commonly used anti-convulsant drugs were examined for the first time on the cloned human type IIA channel. Lamotrigine and phenytoin produced concentration- and voltage-dependent inhibition of the type IIA currents, whereas, sodium valproate and gabapentin (up to 1 mM) had no effect. These results indicate that recombinant human type IIA Na+ channels conduct tetrodotoxin-sensitive Na+ currents with similar properties to those observed in recombinant rat brain type IIA and native rat brain Na+ channels. This stable cell line should provide a useful tool for more detailed characterization of therapeutic modulators of human Na+ channels.

Publication types

  • Comparative Study

MeSH terms

  • Animals
  • Brain Chemistry*
  • CHO Cells
  • Cell Line
  • Cloning, Molecular
  • Cricetinae
  • Electric Conductivity
  • Electrophysiology
  • Gene Expression*
  • Humans
  • Lamotrigine
  • Patch-Clamp Techniques
  • Phenytoin / pharmacology
  • Rats
  • Recombinant Proteins / metabolism
  • Sodium Channels / genetics*
  • Sodium Channels / physiology*
  • Tetrodotoxin / pharmacology
  • Transfection
  • Triazines / pharmacology

Substances

  • Recombinant Proteins
  • Sodium Channels
  • Triazines
  • Tetrodotoxin
  • Phenytoin
  • Lamotrigine