Ca-dependence of synaptic vesicle exocytosis and endocytosis at the hippocampal mossy fibre terminal

Abstract

Key points: We used presynaptic capacitance measurements at the hippocampal mossy fibre terminal at room temperature to measure Ca-dependence of exo- and endocytotic kinetics. The readily releasable pool (RRP) of synaptic vesicles was released with a time constant of 30–40 ms and was sensitive to Ca buffers, BAPTA and EGTA. Our data suggest that recruitment of the vesicles to the RRP was Ca-insensitive and had a time constant of 1 s. In addition to the RRP, the reserve pool of vesicles, which had a similar size to RRP, was depleted during repetitive stimulation. Our data suggest that synaptic vesicle endocytosis was also Ca-insensitive. Abstract: Hippocampal mossy fibre terminals comprise one of the cortical terminals, which are sufficiently large to be accessible by patch clamp recordings. To measure Ca-dependence of exo- and endocytotic kinetics quantitatively, we applied presynaptic capacitance measurements to the mossy fibre terminal at room temperature. The time course of synaptic vesicle fusion was slow, with a time constant of tens of milliseconds, and was sensitive to Ca buffers EGTA and BAPTA, suggesting a loose coupling between Ca channels and synaptic vesicles. The size of the readily-releasable pool (RRP) of synaptic vesicles was relatively insensitive to Ca buffers. Once the RRP was depleted, it was recovered by a single exponential with a time constant of ∼1 s independent of the presence of Ca buffers, suggesting Ca independent vesicle replenishment. In addition to the RRP, the reserve pool of vesicles was released slowly during repetitive stimulation. Endocytosis was also insensitive to Ca buffers and had a slow time course, excluding the involvement of rapid vesicle cycling in vesicle replenishment. Although mossy fibre terminals are known to have various forms of Ca-dependent plasticity, some features of vesicle dynamics are robust and Ca-insensitive.