Figure 3
The function of KCC2 was impaired before the epileptiform bursting activities in vitro. (A,B) Representative traces showing evoked GABAergic currents were recorded at either −80 mV (A left) or −40 mV (B left) during HFS (1.2 s, 20 Hz) under control and CTZ conditions. Line chart showing the synaptic depression with little change under −80 mV condition (A right), but significantly impaired under −40 mV condition (B right). (C,D) E GABA, determined from same neuron in hippocampal slices at either 0 and 0.5 hr (C) or 0 and 1 hr (D) with 50 μM CTZ treatment, was plotted and the mean value was shown in red. No obvious change of E GABA after 0.5 hr (P = 0.70), but a significant depolarizing shift of E GABA after 1 hr CTZ treatment (*P < 0.05) was observed. (E,F) Example recordings of E GABA by gramicidin perforated whole cell patch clamp after 2 hr incubation of either DMSO or 50 μM CTZ. Dot plot indicated that E GABA was significantly positive shifted after 2 hr treatment of CTZ (***P < 0.001). (G) I-V plots showing the experimental determination of the E GABA after either CTZ, furosemide or bumentanide treatment under whole cell recording condition. (H) Bar histogram showing that a significantly depolarization shifts of E GABA with either CTZ (***P < 0.001) or furosemide (*P < 0.05), but not with bumentanide treatment.
