Kinetic organization of Ca2+ signals that regulate synaptic release efficacy in sympathetic neurons.

Calcium regulation of neurotransmitter release is essential for maintenance of synaptic transmission. However, the temporal and spatial organization of Ca(2+) dynamics that regulate synaptic vesicle (SV) release efficacy in sympathetic neurons is poorly understood. Here, we investigate the N-type Ca(2+) channel-mediated kinetic structure of Ca(2+) regulation of cholinergic transmission of sympathetic neurons. We measured the effect of Ca(2+) chelation with fast 1,2-bis(2-aminophenoxy) ethane-tetraacetic acid (BAPTA) and slow ethyleneglycol-tetraacetic acid (EGTA) buffers on exocytosis, synaptic depression, and recovery of the readily releasable vesicle pool (RRP), after both single action potential (AP) and repetitive APs. Surprisingly, postsynaptic potentials peaking at ~12 milliseconds after the AP was inhibited by both rapid and slow Ca(2+) buffers suggests that, in addition to the well known fast Ca(2+) signals at the active zone (AZ), slow Ca(2+) signals at the peak of Ca(2+) entry also contribute to paired-pulse or repetitive AP responses. Following a single AP, discrete Ca(2+) transient increase regulated synaptic depression in rapid (<30-millisecond) and slow (<120-millisecond) phases. In contrast, following prolonged AP trains, synaptic depression was reduced by a slow Ca(2+) signal regulation lasting >200 milliseconds. Finally, after an AP burst, recovery of the RRP was mediated by an AP-dependent rapid Ca(2+) signal, and the expansion of releasable SV number by an AP firing activity-dependent slow Ca(2+) signal. These data indicate that local Ca(2+) signals operating near Ca(2+) sources in the AZ are organized into discrete fast and slow temporal phases that remodel exocytosis and short-term plasticity to ensure long-term stability in acetylcholine release efficacy.