Abstract
Not only the amplitude but also the time course of a presynaptic Ca2+ transient determine multiple aspects of synaptic transmission. In small bouton-type synapses, the mechanisms underlying the Ca2+ decay kinetics have not been fully investigated. Here, factors that shape an action-potential-evoked Ca2+ transient were quantitatively studied in synaptic boutons of neocortical layer 5 pyramidal neurons. Ca2+ transients were measured with different concentrations of fluorescent Ca2+ indicators and analyzed based on a single-compartment model. We found a small endogenous Ca2+-binding ratio (7 ± 2) and a high activity of Ca2+ transporters (0.64 ± 0.03 ms-1), both of which enable rapid clearance of Ca2+ from the boutons. However, contrary to predictions of the single-compartment model, the decay time course of the measured Ca2+ transients was biexponential and became prolonged during repetitive stimulation. Measurements of [Ca2+]i along the adjoining axon, together with an experimentally constrained model, showed that the initial fast decay of the Ca2+ transients predominantly arose from the diffusion of Ca2+ from the boutons into the axon. Therefore, for small boutons en passant, factors like terminal volume, axon diameter, and the concentration of mobile Ca2+-binding molecules are critical determinants of Ca2+ dynamics and thus Ca2+-dependent processes, including short-term synaptic plasticity.