FIGURE 5. Effect of blockade of neuronal VEGF signaling on the neurotrophic and synaptogenic effects of ketamine^a.

^a(A) Experimental timeline for dendritic morphology in rat primary cultured cortical neurons. (B) The number of dendritic crossings at 50 and 100 μm distances from the soma after 24-h vehicle, ketamine (500 nM) or VEGF₁₆₄ (50 ng/mL) treatment with 30-min pre-incubation of 0.1% DMSO with or without ZM323881 (10 nM; 50 μm, interaction, F_2,128 = 11.0, p < 0.0001; 100 μm, interaction, F_2,128 = 17.2, p < 0.0001, n = 15–26). (C) Representative images of EGFP-expressing rat primary cortical neurons from each group with concentric circles (100- and 200-μm diameter). (D) Experimental timeline for dendritic spine analysis. (E) Spine density was analyzed on primary (I) and secondary (II) branches of the apical tuft of layer V pyramidal neurons in the infralimbic (IL; n = 11–15 branches from 6 cells from 3 mice/group) and prelimbic (PL; n =10–17 branches from 6 cells from 3 mice/group) mPFC. Bottom, representative image of a Golgi-stained layer V pyramidal neuron. (F) Left, spine density of infralimbic pyramidal neurons (I, interaction, F_1,45 = 8.45, p = 0.0056, n = 12–13; II, interaction, F_1,49 = 4.06, p = 0.049, n = 11–15). Right, representative images of the apical tuft segments. (G) Left, spine density of prelimbic pyramidal neurons (I, interaction, F_1,43 = 1.51, p = 0.23, main effect of ketamine, F_1,43 = 6.72, p = 0.013, main effect of genotype, F_1,43 = 10.6, p = 0.0022, n = 10–13; II, interaction, F_1,55 = 10.2, p = 0.0023, n = 12–17). Right, representative images of the apical tuft segments. Data are expressed as means ± SD. **p < 0.01, ***p < 0.001; ^#p < 0.05, ^##P < 0.01, ^###P < 0.001 relative to Control+Sal mice.