Blockade of ion channel expression in Xenopus oocytes with complementary DNA probes to Na+ and K+ channel mRNAs

Neuron. 1988 Dec;1(10):963-71. doi: 10.1016/0896-6273(88)90153-5.

Abstract

Ionic currents were recorded from Xenopus oocytes injected with RNA isolated from chick or mouse brain. Three currents were studied: a rapid tetrodotoxin-sensitive Na+ current (Ina), an early outward K+ current sensitive to 4-aminopyridine (IA), and an inward current activated by the excitatory amino acid receptor agonist kainate. Oligonucleotides (60-80 bases long) complementary to rat brain Na+ channel sequences were prehybridized to chick brain RNA. These DNA sequences, upon injection into oocytes, specifically inhibited expression of INa relative to IA and the kainate-induced current in a dose-dependent manner. By contrast, prehybridization of oligonucleotides complementary to sequences either from the Drosophila Shaker locus (which codes for an early K+ current in Drosophila muscle) or from a homologous clone from mouse brain did not block the expression of the early outward K+ current induced in the oocytes by mRNA from chick or mouse brain. This method provides a convenient means for testing the functional role of cloned DNA species.

MeSH terms

  • Animals
  • Base Sequence
  • Cell Membrane / metabolism
  • Cell Membrane / physiology
  • Cell Membrane Permeability / drug effects
  • Cell Membrane Permeability / physiology
  • DNA / genetics
  • DNA / pharmacology
  • DNA Probes*
  • Drosophila / genetics
  • Electric Conductivity
  • Female
  • Genes, Regulator / physiology
  • Kainic Acid / pharmacology
  • Molecular Sequence Data
  • Oocytes / drug effects
  • Oocytes / physiology*
  • Oocytes / ultrastructure
  • Potassium / metabolism
  • Potassium Channels / drug effects*
  • Potassium Channels / physiology
  • Protein Biosynthesis
  • RNA, Messenger / genetics*
  • RNA, Messenger / pharmacology
  • Sodium / metabolism
  • Sodium Channels / drug effects*
  • Sodium Channels / physiology
  • Xenopus / physiology*

Substances

  • DNA Probes
  • Potassium Channels
  • RNA, Messenger
  • Sodium Channels
  • DNA
  • Sodium
  • Potassium
  • Kainic Acid