Action potential counting at giant mossy fiber terminals gates information transfer in the hippocampus

Neuronal communication relies on action potential discharge, with the frequency and the temporal precision of action potentials encoding information. Hippocampal mossy fibers have long been recognized as conditional detonators owing to prominent short-term facilitation of glutamate release displayed during granule cell burst firing. However, the spiking patterns required to trigger action potential firing in CA3 pyramidal neurons remain poorly understood. Here, we show that glutamate release from mossy fiber terminals triggers action potential firing of the target CA3 pyramidal neurons independently of the average granule cell burst frequency, a phenomenon we term action potential counting. We find that action potential counting in mossy fibers gates glutamate release over a broad physiological range of frequencies and action potential numbers. Using rapid Ca²⁺ imaging we also show that the magnitude of evoked Ca²⁺ influx stays constant during action potential trains and that accumulated residual Ca²⁺ is gradually extruded on a time scale of several hundred milliseconds. Using experimentally constrained 3D model of presynaptic Ca²⁺ influx, buffering, and diffusion, and a Monte Carlo model of Ca²⁺-activated vesicle fusion, we argue that action potential counting at mossy fiber boutons can be explained by a unique interplay between Ca²⁺ dynamics and buffering at release sites. This is largely determined by the differential contribution of major endogenous Ca²⁺ buffers calbindin-D_28K and calmodulin and by the loose coupling between presynaptic voltage-gated Ca²⁺ channels and release sensors and the relatively slow Ca²⁺ extrusion rate. Taken together, our results identify a previously unexplored information-coding mechanism in the brain.