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Fig. 5 | BMC Biology

Fig. 5

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

Fig. 5

Effect of cPcdh deficiency on the morphology and synaptic connectivity of clonal neurons. a Barrel structure visualized by cytochrome oxidase reaction (CO) in a chimeric mouse prepared using cPcdh-knockout (KO) induced pluripotent cells. Scale bar: 200 μm. b, c Distribution (b) and proportion (c) of tdTomato-labeled cPcdh-KO cells in the analyzed area in each column. Number of columns is 7, 14, and 10 for P9–11, P13–16, and P18–20, respectively. P = 0.35 (Kruskal–Wallis test). Scale bar: 100 μm. d A cPcdh-deficient spiny stellate neuron (yellow, upper) and trace 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 cPcdh-KO (n = 9 cells sampled from 4 mice; orange) 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 cPcdh-deficient cells and wild-type cells were observed; P = 0.37 (e), P = 0.60 (f), and P > 0.07 (g) (t test). hk Dual whole-cell recordings from cPcdh-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 cPcdh-KO P-P pairs with one-way (upper) or reciprocal (lower) connections. i Percentage of cPcdh-KO P-P pairs with each connection type. Number of recorded pairs is indicated on each bar. Number of mice: P9–11(n = 5), P13–16 (n = 11), P18–20 (n = 9). Developmental changes in connection probability (j) and reciprocity (k) in cPcdh-KO P-P (orange) and wild-type P-P (green) or or GFP-positive and -negative neuron (P-N; gray) cell pairs. *P < 0.05, **P < 0.01, ***P < 0.001 compared with wild-type clonal cell pairs (Fisher’s exact test)

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