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. 2016 Jun 7;3(3):ENEURO.0109-15.2016. doi: 10.1523/ENEURO.0109-15.2016

Figure 1.

Figure 1.

Intracellular Alexa 594 altered the rectification index of spontaneous EPSCs mediated by AMPARs in cerebellar stellate cells. A, Left, Example traces of spontaneous EPSCs recorded at +40 mV (top) and −60 mV (bottom) when 100 µm Alexa 594 was included in the recording electrode. EPSC traces recorded immediately after obtaining the whole configuration (0–5 min) are shown in black, and those recorded at 15–20 min are shown in red. Right, Averaged spontaneous EPSCs. B, RI of sEPSCs recorded with Alexa Fluor 594 in the patch pipette at 0–5 and 15–20 min. Average, bar graphs; individual cells, open circles. Paired t test, p < 0.05 (n = 9 cells, 7 animals). C, Scatter plot of the percentage change in RI at 15–20 min (100 × (RI15–20 min − RI0–5 min)/RI0–5 min) vs the initial RI at 0–5 min of cells filled with Alexa Fluor 594. Stellate cells with a lower initial rectification index have a greater increase in the magnitude of RI. D–G, RI of sEPSC amplitudes recorded at −60 and +40 mV from cells with an initial RI < 0.5 (n = 7). D, RI of EPSCs increased at 15–20 min of recording. Paired t test, p < 0.004. E, The amplitude of sEPSCs at −60 mV did not change. F, sEPSC amplitude at +40 mV increased at 15–20 min. Paired t test: n = 7, p < 0.03. G, The decay time constant of sEPSCs at −60 mV (p = 0.171). H, Time course of average RI (n = 7). One-way repeated-measures ANOVA, p = 0.004; post hoc: Bonferroni t test: 0–5 vs 10–15 min, p < 0.05; 0–5 vs 15–20 min, p < 0.01; n = 7). I, J, Alexa Fluor 594 (100 µm) fluorescence intensity along stellate cell dendrites (5 dendrites, 4 cells, 4 animals) at each time point. I, Plot of the average fluorescence intensity along dendrites from the soma at 0–5 min (open circles) and 15–20 min (red circles). Arrows show the locations where Alexa Fluor 594 fluorescence intensity decreased to the basal level. J, Time course of the diffusion distance of Alexa Fluor 594 along dendrites. One-way repeated-measures ANOVA, p = 0.014; post hoc: Bonferroni t test: 0–5 vs 10–15 min, p < 0.05; 0–5 vs 15–20 min, p < 0.05; n = 5.