Figure 5. Nuclear aggregate formation, morphology and basal PF EPSC in PCs with virally expressed abnormal Ataxin‐1.

A–C, cerebellar slices from mice treated with lentiviral vectors were immunostained 8 weeks after viral injection. Upper and lower panels show native GFP fluorescence and GFP signal overlapped with Calbindin immunofluorescence, respectively. Insets show enlarged PC somas from white‐boxed areas in the corresponding images. Scale bars = 50 μm. PCs expressing lentivirally transduced GFP (A), non‐pathogenic GFP‐ATXN1[Q30] (B) or abnormal GFP‐ATXN1[Q76] (C). D, summary graph showing the percentages of aggregate‐positive PCs. More than 400 PCs from three mice were examined in each condition. ^*** P < 0.0001; Fisher's exact test followed by Holm's multiple pairwise correction. E, representative biocytin‐labelled PC images 8 weeks after injection. Scale bar = 50 μm. F, Sholl analysis to examine the complexity of the PC morphology. Left, schematic representation of the analysis. Sampling concentric shells with 20 μm radius steps are shown for illustration. The PC soma (grey) was excluded from the analysis. Scale bar = 20 μm. Right, the average number of intersections between the dendrites and the concentric shells at different distances from the soma. G, stimulus intensity–EPSC amplitude relationship of PF‐PC EPSCs from non‐injected mice and those that received lentiviral vectors expressing GFP‐ATXN1[Q30] or GFP‐ATXN1[Q76]. Left panel shows representative traces of PF EPSCs elicited by increasing electrical stimulation intensity. The number of PCs examined in each group is shown in parentheses. Two‐way repeated measures ANOVAs indicate no significant difference in the dendritic complexity of PCs (F right, genotype effect, F _1,38 = 0.08, P = 0.77; interaction, F _52,1976 = 0.31, P > 0.99) or the PF EPSC amplitudes (G right, genotype effect, F _2,37 = 0.02, P = 0.98; interaction, F _22,407 = 0.16, P > 0.99). [Colour figure can be viewed at wileyonlinelibrary.com]