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. 2020 Dec 2;40(49):9372–9385. doi: 10.1523/JNEUROSCI.1922-20.2020

Figure 5.

Figure 5.

Quantification of RIM-dependent loss of Munc13-1 activity on synaptic properties. A, Sample traces of current induced by hypertonic sucrose to estimate the RRP size (top), sample traces of EPSCs (middle), and mEPSCs (bottom). Autaptic hippocampal neurons were infected with ΔCre + Scramble (Scr.) shRNA as control, ΔCre + Munc13-1 KD shRNAs (2-40 × 105 IU) to produce Munc13-1 dose gradients, and Cre recombinase + Scramble shRNA to create RIM1/2 cDKO. Dashed lines indicate the maximum current amplitude of RIM1/2 cDKO. B–E, Bar plots represent the normalized mean RRP defined as a charge measured by hypertonic sucrose application (B), normalized EPSC amplitude (C), normalized mEPSC frequency (D), and normalized Pvr (E). F–I, Plots represent the normalized RRP size (F), EPSC amplitude (G), mEPSC frequency (H), and Pvr (I) as a function of normalized Munc13-1/VGLUT1 presynaptic expression. F-I, Black dashed lines indicate fitting with the Hill equation. Blue dashed line indicates Munc13-1/VGLUT1 expression in RIM1/2 cDKO neurons. Data indicate normalized mean ± SEM. *p ≤ 0.05; **p ≤ 0.01; ****p ≤ 0.0001; nonparametric one-way ANOVA with Kruskal–Wallis test followed by Dunn's post hoc test. For the absolute values and statistical overview, see also the table in Extended Data Figure 5-1.