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. 2017 May 30;8:15592. doi: 10.1038/ncomms15592

Figure 6. Autophosphorylation-dependent and -independent roles of Pyk2 in dendritic spines.

Figure 6

(a) Hippocampal neurons from wild-type (WT) and Pyk2 KO mice were cultured for 21–22 days, transfected with plasmids coding GFP or GFP fused to wild-type Pyk2, to Pyk2(1-840), Pyk2(Y402F) or Pyk2-KD (as indicated), and treated with vehicle or glutamate (Glu, 40 μM, 3 h). Neurons were imaged for GFP fluorescence (green) and PSD-95 immunoreactivity (red). (b) Quantification of GFP/PSD-95 double-positive puncta size (that is, yellow puncta) as in a. Two-way ANOVA: interaction, F(6,312)=19.07, P<10−4, glutamate effect, F(1,312)=134.3, P<10−4, Pyk2 expression effect, F(6,312)=20.06, P<10−4. (c) Spine density and length were studied in similar conditions as in a, in the absence of treatment, using GFP or Pyk2:GFP fluorescence. (d) Quantification of spine density. One-way ANOVA: F(6,155)=24.90, P<10−4. (e) Quantification of spine length. One-way ANOVA: F(6,157)=30.68, P<10−4 and. In b,d,e, individual data points and means+s.e.m. are shown, 15–20 dendrites per condition (one–two dendrites per neuron) from two to three independent experiments. Post hoc multiple comparisons were done with Holm-Sidak's test (b,d,e), ***P<0.001, ****P<10−4. Scale bars, 3 μm (a) and 1 μm (c).