Steady-state currents through voltage-dependent, dihydropyridine-sensitive Ca2+ channels in GH3 pituitary cells

Ca2+ influx via voltage-dependent Ca2+ channels is known to be elicited during action potentials but possibly also occurs at the resting potential. The steady-state current through voltage-dependent Ca2+ channels and its role for the electrical activity was, therefore, investigated in pituitary GH3 cells. Applying the recently developed ‘nystatin-modification’ of the patch-clamp technique, most GH3 cells (18 out of 23 cells) fired spontaneous action potentials from a baseline membrane potential of 43.7 ± 4.6 mV (mean ± s. d., n = 23). The frequency of action potentials was stimulated about twofold by Bay K 8644 (100 nM), a Ca2+-channel stimulator, and action potentials were completely suppressed by the Ca2+-channel blocker PN 200—110 (100 nM). Voltage clamping GH3 cells at fixed potentials for several minutes and with 1mM Ba2+ as divalent charge carrier, we observed steady-state Ca2+-channel currents that were dihydropyridine-sensitive and displayed a U-shaped current—voltage relation. The results strongly suggest that the observed long lasting, dihydropyridine-sensitive Ca2+-channel currents provide a steady-state conductivity for Ca2+ at the resting potential and are essential for the generation of action potentials in GH3 pituitary cells.