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Abbreviations used in this paper: ACSF, artificial cerebrospinal fluid; AMC, 7-amino-4-methylcoumarin; AZ, active zone; cryo-ET, cryo–electron tomography; fEPSP, field excitatory postsynaptic potential; HB, homogenization buffer; HBM, Hepes-buffered medium; KO, knockout; K-W, Kruskal-Wallis; PPF, paired-pulse facilitation; RIM, Rab3-interacting molecule; RRP, readily releasable pool; UPS, ubiquitin–proteasome system; WT, wild type