Multiple calcium channel subtypes in isolated rat chromaffin cells

Abstract

By using the whole-cell configuration of the patch-clamp technique we have investigated the pharmacological properties of Ca2+ channels in short-term cultured rat chromaffin cells. In cells held at a membrane potential of --80 mV, using 10 mM Ba2+ as the charge carrier, only high-voltage-activated (HVA) Ca2+ channels were found. Ba2+ currents (IBa) showed variable sensitivity to dihydropyridine (DHP) Ca2+ channel agonists and antagonists. Furnidipine, a novel DHP antagonist, reversibly blocked the current amplitude by 22% and 48%, at 1 microM and 10 microM respectively, during short (15-50 ms) depolarizing pulses to 0 mV. The L-type Ca2+ channel agonist Bay K 8644 (1 microM) caused a variable potentiation of HVA currents that could be better appreciated at low rather than at high depolarizing steps. Increase of IBa was accompanied by a 20-mV shift in the activation curves for Ca2+ channels towards more hyperpolarizing potentials. Application of the conus toxin omega-conotoxin GVIA (GVIA; 1 microM) blocked 31% of IBa; blockade was irreversible upon removal of the toxin from the extracellular medium. omega-Agatoxin IVA (IVA; 100 nM) produced a 15% blockade of IBa. omega-Conotoxin MVIIC (MVIIC; 5 microM) produced a 36% blockade of IBa; such blockade seems to be related to both GVIA-sensitive (N-type) and GVIA-resistant Ca2+ channels. The sequential addition of supramaximal concentrations of furnidipine (10 microM), GVIA (1 microM), IVA (100 nM) and MVIIC (3 microM) produced partial inhibition of IBa, which were additive. Our data suggest that the whole cell IBa in rat chromaffin cells exhibits at least four components.(ABSTRACT TRUNCATED AT 250 WORDS)