Blockage of synaptic release by brief hyperpolarizing pulses in the neuromuscular junction of the crayfish.

1. Synaptic currents were evoked at the neuromuscular junction of the deep extensor abdominal muscle of the crayfish by direct depolarization of motor nerve endings. 2. Quantal content and time course of neurotransmitter release were determined from delay histograms of unitary release events recorded with a macropatch clamp technique. 3. Synaptic facilitation was elicited by pairing depolarizing pulses at intervals ranging from 10 to 200 ms. At 14 degrees C the duration of facilitation was about 50 ms. Reducing activity of the Nao(+)-Cai2+ exchange by lowering [Na+]o by 50% resulted in prolonged facilitation, which lasted approximately 150 ms. 4. Normalized synaptic delay histograms at normal [Na+]o and 50% [Na+]o were the same for the first and the facilitated second response, indicating that activity of the Na(+)-Ca2+ exchange does not determine the time course of release. 5. The application of a hyperpolarizing post-pulse after the first depolarizing stimulus reduced release and altered its time course to a similar extent both in normal and in 50% [Na+]o. However, it did not affect the level and the time course of release of the facilitated response. 6. A hyperpolarizing post-pulse given after the first and second pulses of a pair reduced release to the same extent for the two depolarizing pulses. 7. These results indicate that whereas manipulations thought to increase [Ca2+]i (i.e. reducing activity of the Nao(+)-Cai2+ exchange or facilitation) affect the quantal content, they do not influence the time course of release. However, changes of membrane potential do affect the quantal content, and more importantly the time course of release, thus suggesting a contributory role of membrane potential in the control of synaptic release.