Weak endogenous Ca²⁺ buffering supports sustained synaptic transmission by distinct mechanisms in rod and cone photoreceptors in salamander retina

Differences in synaptic transmission between rod and cone photoreceptors contribute to different response kinetics in rod- versus cone-dominated visual pathways. We examined Ca²⁺ dynamics in synaptic terminals of tiger salamander photoreceptors under conditions that mimicked endogenous buffering to determine the influence on kinetically and mechanistically distinct components of synaptic transmission. Measurements of I_Cl(Ca) confirmed that endogenous Ca²⁺ buffering is equivalent to ~0.05 mmol/L EGTA in rod and cone terminals. Confocal imaging showed that with such buffering, depolarization stimulated large, spatially unconstrained [Ca²⁺] increases that spread throughout photoreceptor terminals. We calculated immediately releasable pool (IRP) size and release efficiency in rods by deconvolving excitatory postsynaptic currents and presynaptic Ca²⁺ currents. Peak efficiency of ~0.2 vesicles/channel was similar to that of cones (~0.3 vesicles/channel). Efficiency in both cell types was not significantly affected by using weak endogenous Ca²⁺ buffering. However, weak Ca²⁺ buffering speeded Ca²⁺/calmodulin (CaM)-dependent replenishment of vesicles to ribbons in both rods and cones, thereby enhancing sustained release. In rods, weak Ca²⁺ buffering also amplified sustained release by enhancing CICR and CICR-stimulated release of vesicles at nonribbon sites. By contrast, elevating [Ca²⁺] at nonribbon sites in cones with weak Ca²⁺ buffering and by inhibiting Ca²⁺ extrusion did not trigger additional release, consistent with the notion that exocytosis from cones occurs exclusively at ribbons. The presence of weak endogenous Ca²⁺ buffering in rods and cones facilitates slow, sustained exocytosis by enhancing Ca²⁺/CaMdependent replenishment of ribbons in both rods and cones and by stimulating nonribbon release triggered by CICR in rods.