L-type calcium channel activity regulates sodium channel levels in rat pituitary GH3 cells

J Physiol. 2000 Feb 15;523 Pt 1(Pt 1):45-55. doi: 10.1111/j.1469-7793.2000.00045.x.

Abstract

1. The effects of chronic pharmacological modulation of L-type Ca2+ channel activity on the cell surface expression of Na+ channels were examined in GH3 cells. 2. Prolonged inhibition (4-5 days) of L-channels with nimodipine caused a 50-60 % decrease in the peak amplitude of whole-cell Na+ currents recorded with the patch-clamp technique. On the contrary, prolonged exposure to the L-channel agonist Bay K 8644 induced an approximately 2.5-fold increase in peak Na+ current. In both cases, there were only minor changes in cell capacitance and no significant changes in Na+ channel gating properties. 3. Measurements of the specific binding of radiolabelled saxitoxin to intact cells showed that nimodipine treatment reduced the number of cell surface Na+ channels, whereas treatment with Bay K 8664 produced the opposite effect. The dual regulation of Na+ channel abundance explained the mentioned changes in Na+ current amplitude. 4. Plasma membrane Na+ channels had a half-life of approximately 17 h both in control cells and in cells treated with Bay K 8644, as estimated from the rate of decay of peak Na+ current after inhibition of protein synthesis with cycloheximide. Actinomycin D, an inhibitor of gene transcription, and also cycloheximide, occluded the stimulatory effect of Bay K 8644 on Na+ current density when measured over a 24 h period. 5. These findings indicate that the entry of Ca2+ through L-type channels influences in a positive way the number of functional Na+ channels in GH3 cells, and suggest that Ca2+ influx stimulates either Na+ channel gene expression or the expression of a regulatory protein that promotes translocation of pre-assembled Na+ channels into the plasma membrane.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester / pharmacology
  • Animals
  • Calcium Channel Agonists / pharmacology
  • Calcium Channel Blockers / pharmacology
  • Calcium Channels, L-Type / physiology*
  • Cell Line
  • Cycloheximide / pharmacology
  • Dactinomycin / pharmacology
  • Electrophysiology
  • Ion Channel Gating / drug effects
  • Nimodipine / pharmacology
  • Nucleic Acid Synthesis Inhibitors / pharmacology
  • Pituitary Gland / cytology
  • Pituitary Gland / metabolism*
  • Protein Synthesis Inhibitors / pharmacology
  • Rats
  • Saxitoxin / metabolism
  • Sodium Channels / metabolism*
  • Sodium Channels / physiology
  • Transcription, Genetic / drug effects

Substances

  • Calcium Channel Agonists
  • Calcium Channel Blockers
  • Calcium Channels, L-Type
  • Nucleic Acid Synthesis Inhibitors
  • Protein Synthesis Inhibitors
  • Sodium Channels
  • Dactinomycin
  • Saxitoxin
  • Nimodipine
  • 3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester
  • Cycloheximide