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Fig. 4 | BMC Biology

Fig. 4

From: Establishment of high reciprocal connectivity between clonal cortical neurons is regulated by the Dnmt3b DNA methyltransferase and clustered protocadherins

Fig. 4

Effect of Dnmt3b deficiency on the morphology and synaptic connectivity of clonal neurons. a Barrel structure visualized by cytochrome oxidase reaction (CO) in a chimeric mouse produced using Dnmt3b-knockout (KO) induced pluripotent stem cells. Scale bar: 200 μm. b, c Distribution (b) and proportion (c) of Dnmt3b-KO cells in the analyzed area in each column. The number of columns is 12, 14, and 13 for P9–11, P13–16, and P18–20, respectively. P = 0.09 (Kruskal–Wallis test). Scale bar: 100 μm. d Dnmt3b-KO spiny stellate cell (red, upper) and traces of its soma and dendrites (lower). Scale bar: 50 μm. eg Total dendritic length (e), number of dendritic branches (f), and Sholl analyses (mean ± SEM, g) of Dnmt3b-KO (purple, n = 9 cells sampled from 4 mice) and wild-type (WT; gray, the same as shown in Fig. 2) spiny stellate cells at P18–20. A bar indicates the mean (e, f). No significant differences in morphological parameters between these two groups were observed; P = 0.41 (e), P = 0.41 (f), and P > 0.14 (g) (t test). hk Dual whole-cell recordings from Dnmt3b-KO GFP-positive excitatory neurons (P-P) pairs. h Representative average (n = 20) traces of presynaptic spikes (pre) and resultant excitatory postsynaptic currents (post) between Dnmt3b-KO P-P pairs with a one-way (upper) or reciprocal (lower) connection. i Percentage of Dnmt3b-KO P-P pairs with one-way (blue), reciprocal (magenta), or no (gray) connections. Numbers of recorded pairs are indicated on the bars. Number of mice: P9–11(n = 9), P13–16 (n = 11), P18–20 (n = 12). *P < 0.05: connection probabilities in different age groups (χ2 test with a 3 × 2 matrix after Bonferroni correction). j, k Connection probability (j) and reciprocity (k) of Dnmt3b-KO P-P (purple) and wild-type P-P (green) or GFP-positive and -negative neuron (P-N) (gray) cell pairs. **P < 0.01 compared with wild-type clonal pairs (Fisher’s exact test)

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