Figure 5.

PF-evoked calcium transients are decreased in patDp/+ Purkinje cell spines. (A) Representative images of a patDp/+ Purkinje cell filled with OGB-2 Ca²⁺ indicator (green) and Alexa 633 dye (red) and visualized spines from that cell. Scale bars = left, 50 μm; right, 2 μm. (B) No difference between resting concentrations of Ca²⁺ between WT and patDp/+ spines (WT, n = 21; patDp/+, n = 11). (C) Representative images of WT (left) and patDp/+ (right) Purkinje cells filled with Fluo-5F Ca²⁺ indicator and Alexa 633 dye. Red box indicates the dendrite with the maximally responding spine. Scale bars = 20 μm (cell images) and 2 μm (dendrite/spine images). (D) Electrophysiological responses from somatic whole-cell patch clamp to two stimulus types: 100-Hz PF burst of 8 pulses alone (PF burst), and a 100-Hz PF burst of 8 pulses + 1 CF pulse (PF burst+CF). (E) Typical traces of simultaneously recorded spine Ca²⁺ transients during each stimulus type. (F) Mean ± SEM traces of Ca²⁺ transients for each stimulus type between WT and patDp/+ spines (WT, n = 14; patDp/+, n = 14). (G) Quantification of Ca²⁺ signal peak amplitudes during a 200-ms window after stimulus onset reveals diminished PF signaling (PF burst and PF burst + CF). ∗p < .05, ∗∗p < .01, ∗∗∗p < .005. Boxplots show mean line with interquartile range. Boxplot error bars show the least extreme of either the highest and lowest values or mean ± (1.5 × interquartile range). Bar plots and average traces show mean ± SEM. All data are shown as median ± SEM. CF, climbing fiber; ns, not significant. OGB, Oregon Green BAPTA; PF, parallel fiber; WT, wild-type.