Re-evaluation of the P/Q Ca2+ channel components of Ba2+ currents in bovine chromaffin cells superfused with solutions containing low and high Ba2+ concentrations

This study was undertaken to reassess the set of voltage-dependent Ca2+ channel subtypes expressed by bovine adrenal chromaffin cells maintained in primary cultures. Previous views on the pharmacology of such channels had to be revised in the light of-the novel data which arose from the use in this study of low and high micromolar concentrations of omega-agatoxin IVA, and low (2 mM) and high (10 mM) concentrations of the charge carrier Ba2+. Whole-cell Ba2+ currents (I-Ba) through Ca2+ channels were elicited in voltage-clamped chromaffin cells, with a holding potential of -80 mV and depolarising pulses to 0 mV. Mean peak I-Ba was 425 pA in 2 mM Ba2+ (59 cells) and 787 pA in 10 mM Ba2+ (42 cells). In 2 mM Ba2+, omega-conotoxin MVIIC (3 mu M) inhibited I-Ba by 79%; in 10 mM Ba2+, the blockade developed much more slowly and reached only 44%. A low concentration of omega-agatoxin IVA (20 nM) inhibited I-Ba by 9%; 2 mu M inhibited I-Ba by 60%. This blockade was similar in low and high Ba2+ concentrations. After giving furnidipine (3 mu M) and omega-conotoxin GVIA (1 mu M), 2 mu M omega-agatoxin IVA inhibited the remaining current (about 40-45%); this blockade was independent of the Ba2+ concentration. The current could be fully blocked by the cocktail furnidipine/omega-conotoxin GVIA/high omega-agatoxin IVA, both in low and high Ba2+ concentrations. The large Q-type channel component of I-Ba is blocked by micromolar concentrations of omega-agatoxin IVA and omega-conotoxin MVIIC. While solutions with a high Ba2+ concentration strongly delayed the development of blockade by omega-conotoxin MVIIC, the blockade by high concentrations of omega-agatoxin IVA was equally effective in solutions with a low or a high Ba2+ concentration. Hence, the use of appropriate Ba2+ and toxin concentrations in this study reveals that P-type Ca2+ channels are poorly expressed in bovine chromaffin cells; in contrast, a robust component of the current depends on Q-type Ca2+ channels. An R-type residual current is not present in these cells.