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. 2020 Jan 7;9:e51822. doi: 10.7554/eLife.51822

Figure 1. Vangl2 affects commissural tracts.

(A) Immunostaining with FluoroMyelin of coronal sections from P21 control (upper panel) and Vangl2 cKO (lower panel) brains in a rostral (left) and more caudal (right) location. Note the absence of the corpus callosum (magenta outline) and hippocampal commissure (cyan outline) caudally in the Vangl2 cKO. Scale bar: 100 µm; n = 3 independent experiments. (B) Cresyl Violet staining of horizontal sections from P31 control (upper panel) and Vangl2 cKO brains (lower panel). Note that the corpus callosum (cc, magenta arrow) and the hippocampal commissure (hc, magenta asterisk) are absent dorsally in Vangl2 cKO brains.Scale bar: 1 mm. (C) Coronal sections from 22‐week‐old control (upper panel) and Vangl2 cKO (lower panel) brains, injected with an AAV‐GFP virus and labeled with DAPI (cyan). Note that hippocampal fibers (in green) cross the cerebral midline in control mice but not in the mice lacking Vangl2. Scale bar: 50 µm. (D) 3D rendering of the hippocampus and the cc covering the hippocampus area. The reconstruction shows the absence of the splenium corpus callosum area in the Vangl2 cKO mouse (white arrow). Abbreviations: hippocampus (hp), corpus callosum (cc), dorso/ventral (D/V), rostrao/caudal (R/C). Scale bars: 800 µm. (E) Cresyl violet staining of 10 week-old control and Vangl2 cKO brains and quantification of cortical thickness of the somato-sensory area. Scale bar: 1 mm; n = 3–5 mice. Data are presented as box-and-whisker plots (min/max) based on three independent experiments. (F) Cux1/DAPI staining of P0 cortices from control and Vangl2 cKO mice. The Cux1 layer is depicted with a dotted magenta line. Scale bar: 100 µm. (G) Quantification analysis show no difference in the number of Cux1 positive cells between P0 control and Vangl2 cKO mice; n = 3 independent experiments. Data are presented as box-and-whisker plots (min/max) based on three independent experiments.

Figure 1.

Figure 1—figure supplement 1. Vangl2 affects commissural tracts.

Figure 1—figure supplement 1.

(A) For Western blots, we collected tissues from at least 3 P0/1 mice for each genotype that express Cre in the cortex and hippocampus (left panel), and tissues that do not express Cre as the cerebellum and spinal cord (right panel). GADPH was used as a loading control in these experiments. (B) Vangl2 immunohistochemistry of E13.5 telencephalon sections. Note the decrease in protein in the dorsal region. Scale bar: 200 µm. (C) Neurofilament (green) and DAPI (blue) staining of E13.5 control and Vangl2 cKO brains show that internal capsule (ic, outlined with magenta arrowheads) does not appear affected by Vangl2 deletion. Scale bar: 100 µm. (D) Hematoxylin staining of coronal sections from P21 control (upper panel) and Vangl2 cKO brains (lower panel). Note that the corpus callosum (cc, black arrow) and the hippocampal commissure (hc, black asterisk) are absent caudally in Vangl2 cKO brains (magenta arrow and asterisk). Scale bar, 20 mm, n = 3 independent experiments. (E) Immunolabeling with Neurofilament-H in horizontal sections from P21 control (upper panel) and Vangl2 cKO (lower panel) brains. Note the absence of the cc (magenta outline) and hc (cyan outline) dorsally in the Vangl2 cKO. Scale bar: 1 mm; n = 3 independent experiments.