Figure 3: Pyk2 localization and dendritic spine density and morphology in Pyk2-deficient mice. | Nature Communications

Figure 3: Pyk2 localization and dendritic spine density and morphology in Pyk2-deficient mice.

From: Pyk2 modulates hippocampal excitatory synapses and contributes to cognitive deficits in a Huntington’s disease model

Figure 3

(a,b) Confocal microscopy images of CA1 stratum radiatum immunostained for (a) Pyk2 (red) and MAP2 (green; inset, higher magnification of the indicated white box) and for (b) Pyk2 (green) and PSD-95 (red; white arrows, double-labelled puncta). Scale bars, 80 μm (a) and 3 μm (b). (c) Electron microscopy in the same region showing of Pyk2 immunoreactive gold particles in a presynaptic terminal (arrow) and a PSD (arrowhead). Scale bar, 0.2 μm. (d) Immunofluorescence PSD-95-positive puncta in the CA1 stratum radiatum from Pyk2+/+ and Pyk2−/− mice. Scale bar, 5 μm. (e) Quantification of puncta as in d. Data are means+s.e.m. (7–10 mice per genotype, three quantified sections per mouse). One-way ANOVA F(2,21)=10.23, P=0.0008. Holm-Sidak’s multiple comparisons test versus +/+, **P<0.01, ***P<0.001. (f) Golgi-Cox-stained apical dendrites of CA1 stratum radiatum pyramidal neuron from Pyk2+/+, Pyk2+/− and Pyk2−/− mice. Scale bar, 3 μm. (g) Quantification of spine density in dendrites as in f, three–four animals per genotype, one-way ANOVA, F(2,146)=14.95, P<10−4 (n=47–54 dendrites per group), post hoc analysis with Holm-Sidak’s multiple comparisons test versus +/+, **P<0.01, ****P<10−4 and −/− versus −/+, °°P<0.01. (h,i) Cumulative probability of spine head diameter (h, n=80) and spine neck length (i, n=115) in ∼60 dendrites from three–four animals per genotype. Distributions were compared with the Kolmogorov–Smirnov test: spine head diameter no significant difference, neck length +/+ versus +/−, D=0.108, P=0.04, +/+ versus −/−, D=0.154, P=0.0005. In e,g, data are means+s.e.m. All mice were 3–4-month old.

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