N-type VGCCs act exclusively through nanodomain signaling. A1, Representative examples of EPSCs recorded in control (black) and in the presence of AgTx + EGTA-AM (blue). A2, EPSC amplitude of tightly coupled P/Q-type VGCCs (open blue circles) compared with the EPSC amplitude generated by the total population of P/Q-type VGCCs (red square). B1, Example of EPSCs recorded in control condition (black) and following the application of CTx + EGTA-AM (red). B2, Graph showing the EPSC amplitude generated by tightly coupled N-type VGCCs (open red circles) compared with the EPSC amplitude evoked by all N-type VGCCs during trains of stimuli. C, Percentage of effect of toxins, EGTA-AM, and combined application of compounds on the 1st and 10th EPSCs generated in trains. Coapplication of CTx and EGTA-AM prevents the decrease in the percentage of effect of CTx during trains. Data represent mean ± SEM. **p < 0.01.