Transient Confinement of Ca_V2.1 Ca²⁺-Channel Splice Variants Shapes Synaptic Short-Term Plasticity

The precision and reliability of synaptic information transfer depend on the molecular organization of voltage-gated calcium channels (VGCCs) within the presynaptic membrane. Alternative splicing of exon 47 affects the C-terminal structure of VGCCs and their affinity to intracellular partners and synaptic vesicles (SVs). We show that hippocampal synapses expressing VGCCs either with exon 47 (Ca_V2.1₊₄₇) or without (Ca_V2.1_Δ47) differ in release probability and short-term plasticity. Tracking single channels revealed transient visits (∼100 ms) of presynaptic VGCCs in nanodomains (∼80 nm) that were controlled by neuronal network activity. Surprisingly, despite harboring prominent binding sites to scaffold proteins, Ca_V2.1₊₄₇ persistently displayed higher mobility within nanodomains. Synaptic accumulation of Ca_V2.1 was accomplished by optogenetic clustering, but only Ca_V2.1₊₄₇ increased transmitter release and enhanced synaptic short-term depression. We propose that exon 47-related alternative splicing of Ca_V2.1 channels controls synapse-specific release properties at the level of channel mobility-dependent coupling between VGCCs and SVs.