Effects of Bay K 8644 on spontaneous and evoked transmitter release at the mouse neuromuscular junction

The dihydropyridine, Bay K 8644, was applied in vitro to mouse phrenic nerve-diaphragm muscle preparations. The drug increased both spontaneous and evoked release of acetylcholine from the motor nerve terminal in a concentration- and time-dependent manner. The rise in miniature endplate potential frequency, however, was the result of an increased intraterminal mobilization of free calcium, rather than well-established activation of voltage-dependent calcium channels. This view is supported by the following observations: (1) an increase in frequency was apparent in Ca2+-free medium; (2) Bay K 8644 is known to require a moderate depolarization to affect Ca2+ channels, but no membrane depolarization was detected; and (3) exposure to low Ca2+ and high Mg2+ medium did not diminish the effect on miniature endplate potential frequency. In a medium containing low Ca2+ and high Mg2+, Bay K 8644 increased quantal content of the evoked endplate potentials to a greater degree and with a faster time course than the frequency of miniature endplate potentials. This enhancement in evoked release did not appear to be caused solely by an increase in cytoplasmic Ca2+, but rather reflected at least in part the Bay K 8644-induced activation of voltage-gated Ca2+ channels, perhaps L-type, at the presynaptic nerve terminal. Thus, we propose that Bay K 8644 exerts dual effects on the motor nerve endings, characterized by a primary action on the presynaptic Ca2+ channels and a secondary action associated with the elevation of intracellular Ca2+ concentration.