Figure 8

Menin knockdown reduces nicotine-induced presynaptic facilitation. (A,B) Representative patch clamp traces recorded from GFP+ hippocampal pyramidal neurons at DIV 10–14 (n ≥ 15, 3 independent experiments, representative traces). The mean frequency and amplitude of mEPSCs was analyzed before (Pre-nicotine, 10 s; top trace) and after (Post-nicotine, 10 s; bottom trace) the nicotine pulse (10 μM, 250 ms, 10 PSI; not shown). nAChR-mediated increase in mEPSC frequency (e.g. arrow) is observed in a GFP+ neuron from NTC-shRNA expressing cultures (A), but is absent in a GFP+ neuron from MEN1-shRNA expressing cultures (B). (C) Summary data, incidence of nicotine-induced presynaptic facilitation in neurons from untreated control (n = 13/19), NTC shRNA (n = 11/15; GFP+) and MEN1 shRNA (n = 5/17; GFP+) transduced hippocampal cultures (post/pre-nicotine relative mEPSC frequency ≥1 ± SEM; see Fig. S5 and Table S8). *Statistical significance (Chi-squared test), P < 0.05. (D) Summary data, mean frequency of mEPSCs before (pre) and after (post) the nicotine pulse in the population of neurons from untreated control (n = 13), NTC shRNA (n = 11; GFP+) and MEN1 shRNA (n = 5; GFP+) transduced cultures that exhibit nicotine-induced presynaptic facilitation. ***Statistical significance (paired t-test), P < 0.001. (E) Summary data, mean amplitude of mEPSCs, as in (D). See also Tables S7–S9 and Figs S5–S7.