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Abstract:
A newly developed deconvolution method (Neher and Sakaba, 2001) allowed us to resolve the time course of neurotransmitter release at the calyx of Held synapse and to quantify some basic aspects of transmitter release. First, we identified a readily releasable pool (RRP) of synaptic vesicles. We found that the size of the RRP, when tested with trains of strong stimuli, was constant regardless of the exact stimulus patterns, if stimuli were confined to a time interval of ∼60 msec. For longer-lasting stimulus patterns, recruitment of new vesicles to the RRP made a substantial contribution to the total release. Second, the cooperativity of transmitter release as a function of Ca2+ current was estimated to be 3–4, which confirmed previous results (Borst and Sakmann, 1999; Wu et al., 1999). Third, an initial small Ca2+ influx increased the efficiency of Ca2+ currents in subsequent transmitter release. This type of facilitation was blocked by a high concentration of EGTA (0.5 mm). Fourth, the release rates of synaptic vesicles at this synapse turned out to be heterogeneous: once a highly Ca2+-sensitive population of vesicles was consumed, the remaining vesicles released at lower rates. These components of release were more clearly separated in the presence of 0.5 mm EGTA, which prevented the buildup of residual Ca2+. Conversely, raising the extracellular Ca2+ concentration facilitated the slower population such that its release characteristics became more similar to those of the faster population under standard conditions. Heterogeneous release probabilities are expected to support the maintenance of synaptic transmission during high-frequency stimulation.
