Monitoring of Ca(2+)-transients in electrically stimulated A7r5 vascular smooth muscle cells fills the experimental gap between KCL-induced depolarization and patch-clamp studies

J Pharmacol Toxicol Methods. 1993 Nov;30(3):143-7. doi: 10.1016/1056-8719(93)90037-f.

Abstract

The effects of electrical field stimulation on [Ca2+]i in the A7r5 vascular smooth muscle cell line have been monitored with the Ca(2+)-sensitive dye fura-2. The experimental set-up allowed high-temporal resolution of the [Ca2+]i-measurements and fast application of test solutions. Electrical field stimulation of A7r5 cells in the confluent growth state resulted in a transient increase in [Ca2+]i from resting values below 100 nM to values in the range of some hundred nM. For a given cell, the electrically induced Ca(2+)-transients were highly reproducible. The requirement for the presence of extracellular Ca2+ and the sensitivity to the Ca(2+)-antagonist nifedipine and the Ca(2+)-agonist BAY K 8644 suggest that the Ca(/+)-transients reflect [Ca2+]i-changes based on Ca(2+)-influx through voltage-dependent L-type Ca(2+)-channels. Therefore, electrical field stimulation of confluent A7r5 cells provides an easy-to-establish and highly reproducible method for the investigation of the physiology and pharmacology of voltage-dependent Ca(2+)-channels in intact vascular smooth muscle cells, which fills the gap between KCl-induced depolarization and the patch-clamp technique.

MeSH terms

  • Animals
  • Calcium / physiology*
  • Calcium Channels / physiology
  • Cells, Cultured
  • Electric Stimulation
  • Electrophysiology / methods
  • Ion Transport
  • Muscle, Smooth, Vascular / cytology
  • Muscle, Smooth, Vascular / physiology*
  • Potassium Chloride
  • Rats
  • Reproducibility of Results

Substances

  • Calcium Channels
  • Potassium Chloride
  • Calcium