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. 2021 Jan 17;296:100302. doi: 10.1016/j.jbc.2021.100302

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© 2021 The Authors

This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

PMC Copyright notice

100-μM 3,4-DAP dose dependently increases neuromuscular transmission independent of Cav1 channels in frog neuromuscular junctions.A and B, sample traces of electrophysiological recordings of EPPs (A) and mEPPs (B) before and after 100-μM 3,4-DAP application. C, quantified quantal content before (circles) or after (triangles) 100-μM 3,4-DAP application to vehicle (pink) or nitrendipine (blue) treated frog NMJs. Two-way mixed ANOVA was used (there was a significant main effect of 100-μM 3,4-DAP (F (1, 19) = 31.66, ∗∗∗p < 0.0001; no significant main effect of nitrendipine (F (1,19) = 4.226, p = 0.0538) nor a significant interaction between 3,4-DAP x nitrendipine (F (1,19) = 4.162, p = 0.0555): vehicle, n = 13; nitrendipine n = 8). D and E, plots of individual paired values (gray dotted lines) with a superimposed average (solid pink line, vehicle; solid blue line, nitrendipine). F, the 100-μM dose of 3,4-DAP increased mEPP frequency in both the vehicle (pink) and nitrendipine (blue) conditions; two-way mixed ANOVA (significant main effect of 3,4-DAP; F (1,19) = 9.541, p = 0.006; no main effect of nitrendipine; F (1,19) = 0.0264, p = 0.8727) or a significant interaction between 3,4-DAP and nitrendipine; F (1,19) = 0.0568, p = 0.8143). G, the 100-μM dose of 3,4-DAP significantly altered mEPP amplitude in both the vehicle (pink) and nitrendipine (blue) conditions; two-way mixed ANOVA (significant main effect of 3,4-DAP; F (1,19) = 11.88, p = 0.0027, but not nitrendipine; F (1,19) = 0.3468, p = 0.5628, and no significant interaction between 3,4-DAP and nitrendipine; F (1,19) = 1.436, p = 0.2454). H and I, the 100-μM dose of 3,4-DAP increased EPP amplitude, shown as individual pairs (gray dotted lines) with a superimposed average (solid pink line, vehicle, H; solid blue line, nitrendipine, I); paired t test, ∗∗p = 0.0012, ∗∗∗p < 0.0001. 3,4-DAP, 3,4-diaminopyridine; Cav, voltage-gated calcium; EPP, endplate potential; mEPP, miniature EPPs; NMJ, neuromuscular junction.