Fig. 5
De novo mutations in TAOK2 impair dendrite growth and synaptic connectivity in the mouse cortex. a Left: representative images of WT or Het Taok2 cortical neurons from P21 mice in utero electroporated at E15 with GFP only (Venus, control) or Het Taok2 cortical neurons with TAOK2α/β, TAOK2α/β A135P, and TAOK2β P1022*. Scale bars represent 20 μm. Right: dendritic heat maps of superimposed neuron tracings for each condition. Blue to yellow indicates increased probability of dendrite presence. Scale bars represent 40 μm. (b) Left: Het Taok2 neurons show reduced dendritic complexity that can be partially rescued with overexpression of TAOK2α/β in the Het background (WT + Venus = 20 cells, Het + Venus = 18 cells, Het + TAOK2α/β = 16 cells from three different brains; two-way ANOVA, post hoc Tukey’s test; F2, 1989 = 42.48, p < 0.0001 between genotypes; *represents ranges of significance; WT + Venus vs. Het + Venus (blue), WT + Venus vs. Het + TAOK2α/β (gray), Het + Venus vs. Het + TAOK2α/β (gray); see supplemental statistics). Right: TAOK2α/β A135P reduces proximal dendritic complexity and TAOK2β P1022* enhances distal dendritic complexity compared with TAOK2α/β (Het + Venus = 20 cells, Het + TAOK2α/β = 16 cells, Het + TAOK2α/β A135P = 20 cells and Het + TAOK2β P1022 = 16 cells from three different brains; two-way ANOVA, post hoc Dunnett’s test; F3, 2720 = 184.1, p < 0.0001 between genotypes; *represents ranges of significance; TAOK2α/β vs. Venus (gray), TAOK2α/β vs. A135P (red), TAOK2α/β vs. P1022* (green); see supplemental statistics). c Dendritic spine distribution map of cortical neurons from WT Taok2 cortical neurons from P21 mice in utero electroporated at E15 d TAOK2α/β A135P and TAOK2β P1022* reduce the number of spines on proximal dendrites, whereas TAOK2β P1022* also shifts spines to distal dendrites (Venus = 11414, TAOK2α/β = 8859, α/β A135P = 9071 and β P1022* = 8686 spines from six neurons from three different brains per condition; two-way ANOVA, post hoc Tukey’s test; F3, 780 = 19.58, p < 0.0001 between genotypes; *represents ranges of significance; Venus vs. TAOK2α/β (gray), T AOK2α/β vs. β P1022*(green), Venus vs. α/β A135P (red), Venus vs. β P1022*(green); see supplemental statistics). (e) TAOK2α/β A135P and TAOK2β P1022* show no significant change in dendritic spines per neuron (Venus = 11,414, TAOK2α/β = 8859, α/β A135P = 9071 and β P1022* = 8686 spines from six neurons from three different brains per condition; spine number: one-way ANOVA, post hoc Dunnett’s; F3, 20 = 1.539, p = 0.2354). f TAOK2α/β A135P significantly increases long thin spines and reduces mushroom-like spines in Taok2 Het neurons (Venus = 11,414, TAOK2α/β = 8859, α/β A135P = 9071 and β P1022* = 8686 spines from six neurons from three different brains per condition; two-way ANOVA, post hoc Dunnett’s test; F3, 120 = 4.36e-13, p > 0.9999; Long Thin: TAOK2α/β vs. TAOK2 α/β A135P p = 0.0001; Mushroom: TAOK2α/β vs. TAOK2 α/β A135P p = 0.0003). *p < 0.05, ***p < 0.001. Values are mean ± s.e.m.
