Neocortical High Probability Release Sites Are Formed by Distinct Ca²⁺ Channel-to-Release Sensor Topographies during...

Coupling distances between Ca²⁺ channels and release sensors regulate vesicular release probability (p_v). Tight coupling is thought to provide a framework for high p_v and loose coupling for high plasticity at low p_v. At synapses investigated during development, coupling distances decrease, thereby increasing p_v and transmission fidelity. We find that neocortical high-fidelity synapses deviate from these rules. Paired recordings from pyramidal neurons with "slow" and "fast" Ca²⁺ chelators combined with experimentally constrained simulations suggest that coupling tightens significantly during development. However, fluctuation analysis revealed that neither p_v (∼0.63) nor the number of release sites (∼8) changes concomitantly. Moreover, the amplitude and time course of presynaptic Ca²⁺ transients are not different between age groups. These results are explained by high-p_v release sites with Ca²⁺ microdomains in young synapses and nanodomains in mature synapses. Thus, at neocortical synapses, a developmental reorganization of the active zone leaves p_v unaffected, emphasizing developmental and functional synaptic diversity.