Fig. 7

WNT5A secreted by HbChP controls morphogenesis of embryonic cerebellum. a In situ analysis of Wnt5a and Wls expression in the dorsal neuroepithelium adjacent to HbChP at E13.5 shows transcripts restriction to the HbChP epithelium, n = 4. Lower panel highlights absence of Wnt5a and low Wls expression in cerebellar ventricular zone (cVZ, empty arrowheads) compared to the HbChP (arrowheads). Scale bar: top 50 µm, bottom 10 μm. b WNT5A observed in the apical cVZ at E14.5 (arrowheads) is absent in Wnt5a^KO embryos (empty arrowheads), n = 3. Scale bar: top 5 μm, inset 2 μm. c WNT5A and APOE (arrowheads) or APOE with APOJ (empty arrowhead) immunostaining in the apical cVZ (dotted line) at E14.5, n = 4. Scale bar: top 5 μm, bottom 2 μm. (d) Scheme of analyzed cerebellar parameters: total area (dash line), width (latero-medial, L-M, horizontal arrows) and length (dorso-ventral, D-V, vertical arrows). Scale bar: 200 μm. e Coronal sections of cerebellum in Wnt5a^WT and Wnt5a^KO embryos. Scale bar: 200 μm. f tdTomato staining in E14.5 FoxJ-CreERT2 embryo demonstrates recombination in HbChP epithelium, which is absent in the cVZ (inset, empty arrowhead) compared to HbChP (arrowheads), n = 3. Scale bar: top 50 µm, inset 20 µm. g Coronal sections of cerebellum in Wnt5a^WT vs Wnt5a^cKO embryos. Scale bar: 200 μm. h–k Analysis of the h total area, i length, j width j, and k width/length ratio of cerebellum in Wnt5a^WT, Wnt5a^KO and Wnt5a^cKO embryos. Graphs show individual data points (dots) from n = 3 biologically independent samples; error bars represent mean ± s.d.; P-values (two-tailed Student’s t-test with unequal variance): * P < 0.05, ** P < 0.01, **** P < 0.0001. Corresponding P-values for differences between Wnt5a^WT (WT), Wnt5a^KO (KO) and Wnt5a^cKO (cKO)—h: WT vs. KO: 0.0004, WT vs. cKO: 0.0001; i WT vs KO: 0.0353, WT vs cKO: 0.0098; j WT vs. KO or cKO: – not significant; k WT vs KO: 0.016, WT vs cKO: 0.0115. l Schematic depiction of the model for WNT5A secretion by HbChP and its transventricular delivery to recipient regions. Source data are provided as a Source Data file