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. 2017 Oct 20;28(1):90–110. doi: 10.1038/cr.2017.128

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Copyright © 2017 The Author(s)

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Abnormal PC firing of Prrt2-deficient mice. (A) Schematic diagrams for the generation of GluN2C-iCre;Ai32;Prrt2^−/− mice. (B) IHC results showed specific expression of the ChR2-YFP fusion proteins in all GCs (white) in cerebellar cortex, but not in Pvalb-positive PCs and other GABAergic interneurons (red). Scale bar, 400 μm. (C) Western blotting shows complete depletion of PRRT2 protein in GluN2C-iCre;Ai32;Prrt2^−/− mouse cerebellum (−/−). GluN2C-iCre;Ai32;Prrt2^+/+ mice were used as the control (+/+). (D) Spontaneous firing activity recorded from PCs on GluN2C-iCre;Ai32;Prrt2^+/+ and GluN2C-iCre;Ai32;Prrt2^−/− mouse cerebellar slices. Upper: representative traces. The first 1-s segments indicated by lines in the upper panels are shown at higher magnification below. Lower: statistical analysis of average firing rate. Each mark in the scatter plots represents individual average firing properties. Bars indicate mean ± SEM. (E) Spontaneous firing activity of PCs after optogenetic stimulation (blue bar). Upper: representative traces. Lower: summary of spontaneous firing frequency. Black, n = 40 cells from 8 GluN2C-iCre;Ai32;Prrt2^+/+ mice. Red, n = 30 cells from 7 GluN2C-iCre;Ai32;Prrt2^−/− mice. Black asterisks on the top indicate statistical significance at individual time points. (F) Cell-attached recording of PCs in the absence or presence of GABAa receptor blocker GABAzine. (G) Effect of glutamate receptor blocker NBQX and AP5 on PC firing activity after optogenetic stimulation.