The nanoscale mobility of calcium channels is driven by readily releasable synaptic vesicles to support precise neurotransmission in live <i>C. elegans</i>

, we report that two diffusive behaviors of N-type VGCCs at AZs are governed by primed SVs via distinct mechanisms. VGCCs diffuse slowly in multiple nanodomains (~100 nm) that spatially correlate with UNC-10/RIM, and the channels' mobility and nanodomain numbers are coupled to SNAREs assembly and SV priming levels via RAB-3/UNC-10/UNC-13L tripartite complexes. Outside nanodomains, a faster mobility of VGCCs and their AZ distribution scale with UNC-13S-mediated priming under an antagonistic regulation by UNC-10 and TOM-1/Tomosyn. These findings reveal unexpected mechanisms whereby pools of readily releasable SVs actively control the nanoscale mobility and nanodomain organization of VGCCs to support precise neurotransmission.