Studies on maitotoxin-induced intracellular Ca(2+) elevation in chinese hamster ovary cells stably transfected with cDNAs encoding for L-type Ca(2+) channel subunits

J Pharmacol Exp Ther. 1999 Aug;290(2):725-30.

Abstract

The aim of the present study was to characterize the role played by different L-type Ca(2+) channel subunits in [Ca(2+)](i) increase induced by maitotoxin (MTX). In the presence of 5 mM extracellular K(+), MTX (0.01-0.5 ng/ml) induced a significant concentration-dependent increase in Fura-2-monitored [Ca(2+)](i) in single Chinese hamster ovary (CHO) cells expressing the alpha(1c) (CHOCalpha9 cells) or the alpha(1c)beta(3)alpha(2)delta (CHOCalpha9beta3alpha2/delta4 cells) subunits of voltage-gated Ca(2+) channels (VGCCs), whereas the effect was much reduced in wild-type CHO cells lacking VGCCs. In addition, MTX effect on CHOCalpha9, CHOCalpha9beta3alpha2/delta4, and GH(3) cells (0.01-0.1 ng/ml) was inhibited by the selective L-type Ca(2+) channel entry-blocker nimodipine (10 microM); a nimodipine-insensitive component was still present, particularly at high (>1 ng/ml) toxin concentrations. In CHOCalpha9beta3alpha2/delta4 cells, depolarizing concentrations of extracellular K(+) (55 mM) reinforced the [Ca(2+)](i) increase induced by MTX (0.1 ng/ml), and this effect was prevented by nimodipine (10 microM). Finally, patch-clamp experiments in CHOCalpha9beta3alpha2/delta4 cells showed that low MTX concentrations (0.03 ng/ml) induced the occurrence of an inward current at -60 mV, which was completely prevented by Cd(2+) (100 microM) and by nimodipine (10 microM), whereas the same dihydropyridine concentration (10 microM) failed to prevent the electrophysiological effects of a higher toxin concentration (3 ng/ml). In conclusion, the results of the present study showed that MTX-induced [Ca(2+)](i) elevation involves two components: 1) an action on L-type VGCCs at the pore-forming alpha(1c) subunit level, which is responsible for the greatest rise of [Ca(2+)](i); and 2) a VGCC-independent mechanism that is present both in excitable and in nonexcitable cells and is responsible for a lower elevation of [Ca(2+)](i).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Barium / metabolism
  • CHO Cells
  • Calcium / metabolism*
  • Calcium Channel Blockers / pharmacology
  • Calcium Channels / biosynthesis*
  • Calcium Channels / genetics
  • Calcium Channels, L-Type
  • Cricetinae
  • DNA, Complementary / genetics
  • Extracellular Space / drug effects
  • Extracellular Space / metabolism
  • Fluorescent Dyes
  • Fura-2
  • Ion Channel Gating
  • Marine Toxins / toxicity*
  • Nimodipine / pharmacology
  • Oxocins*
  • Patch-Clamp Techniques
  • Potassium / pharmacology
  • Transfection

Substances

  • Calcium Channel Blockers
  • Calcium Channels
  • Calcium Channels, L-Type
  • DNA, Complementary
  • Fluorescent Dyes
  • Marine Toxins
  • Oxocins
  • Barium
  • Nimodipine
  • maitotoxin
  • Potassium
  • Calcium
  • Fura-2