Skip to main content
. 2017 May 30;8:15592. doi: 10.1038/ncomms15592

Figure 8. Pyk2 protein levels restoration in the hippocampus partly rescues R6/1 mice phenotype.

Figure 8

(a) Pyk2 and α-tubulin (loading control) immunoblotting in 3-month WT mice injected with AAV-GFP (WT-GFP), or R6/A injected with AAV-GFP (R6/1-GFP) or AAV-Pyk2 and GFP (R6/1-Pyk2). Uncropped blots in Supplementary Fig. 11. (b) Quantification of results as in a (six–nine mice per group). One-way ANOVA: F(2,18)=4.39, P<0.05, Holm-Sidak's test versus R6/1-GFP. (c) Y-maze spontaneous alternation test (10–11 mice per group). Two-way ANOVA interaction F(2,56)=4.39, P<0.05, OA versus NA, Holm-Sidak's test WT-GFP t=2.64, P<0.05, R6/1-GFP, t=0.97, ns, R6/1-Pyk2, t=2.93, P<0.05. (d) NOL test (9–12 mice per group). Two-way ANOVA interaction F(2,54)=11.9, P<0.0001, OL versus NL, Holm-Sidak's test WT-GFP t=9.08, P<0.0001, R6/1-GFP, t=1.60, ns, R6/1-Pyk2, t=6.66, P<0.0001. (e) LTP studied as in Fig. 1c in hippocampal slices from 5-month WT-GFP, R6/1-GFP and R6/1-Pyk2 mice (n=3–4 mice per group, 2–3 slices per mouse, 10–11 slices total). (f) Ten-min average of fEPSP slope 40 min after HFS, normalized to the mean of 10-min baseline (corresponding time points are indicated in e by an horizontal line). Kruskal–Wallis=15.63, P<0.05, post hoc analysis with Dunn's multiple comparisons test. (g) Golgi-Cox-stained apical dendrites from CA1 pyramidal neurons. Scale bar, 3 μm. (h) Quantitative analysis of dendritic spine density as in e (59–62 dendrites from four mice per group). One-way ANOVA: F(2,177)=46.7, P<10−4, Holm-Sidak's test versus R6/1-GFP. (i) Hippocampal sections of WT and R6/1 mice injected with AAV-GFP or AAV-Pyk2 as indicated, and double-immunostained for PSD-95 and Pyk2. High magnification in CA1 stratum radiatum is shown. Scale bar, 5 μm. (j) Quantification of PSD-95-positive puncta density. One-way ANOVA F(2,14)=10.81, P=0.0014, Holm-Sidak's multiple comparisons test. (k) Quantification of PSD-95/Pyk2 double-positive puncta density. One-way ANOVA F(2,14)=9.76, P=0.0022, Holm-Sidak's multiple comparisons test. In j,k, five–seven mice per group. In all graphs values are means+s.e.m. *P<0.05, **P<0.01, ***P<0.001 and ****P<10−4.