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. 2017 Oct 23;7:13847. doi: 10.1038/s41598-017-14185-3

Figure 2.

Figure 2

Expression of Sulf1 and Sulf2 mRNAs and In Vivo Rescue of the CST Defects in Sulf1/2 DKO Mice. (a–f,a’–f’) In situ hybridization of Sulf1 (a–f) and Sulf2 (a’–f’) in the coronal sections of E15.5 brains. Arrowheads show high and overlapping expression of Sulf1 and Sulf2 in the ventricular zone of the third ventricle and aqueduct. Positions of the sections in (a–f) and (a’–f’) are shown in the upper margin. (g–t) Electroporation-mediated rescue of the CST defects in Sulf1/2 DKO mice. The indicated plasmids and pCX-EGFP were electroporated into E12.5 Sulf1/2 DKO brains. At E18.5, EGFP expression and the CST trajectory were examined. Representative results showing electroporation into the medial cerebral cortex (g–j), from the superior colliculus to the cerebellum (k–n), and from the hypothalamus to the midbrain (Mb, o–t) are shown. Figures (q,r) and (s,t) show the electroporated and nonelectroporated sides of the same embryo, respectively. Dorsal (g,k,o), lateral (h,i,l,m,p,q,s), and ventral (j,n,r,t) views are shown. EGFP fluorescence in the hypothalamus in (h) shows transmission through the contralateral side. Arrows indicate the sites of electroporation. Closed arrowheads (i,j,m,n,s,t) indicate abnormal CST fibers in the Sulf1/2 DKO brain, whereas open arrowheads (q,r) indicate the CST restored by electroporation. Statistical analyses of the neurofilament-positive fibers in the midbrain (q,s) are shown in Supplementary Table 1. (u–w) Colocalization of coelectroporated genes. When EGFP and DsRed2 were electroporated together, they were expressed in the same brain regions. Aq, aqueduct; ChP, choroid plexus; CP, cortical plate; cst, corticospinal tract; FM, foramen of Monro; ml, medial lemniscus; OB, olfactory bulb; PoA, preoptic area; preTc, pretectum; Sp, septum; Th; thalamus; V3, third ventricle. Anterior-posterior (A-P), dorsal-ventral (D-V), and medial-lateral (M-L) body axes are shown. Scale bars indicate 300 μm (a,a’), 750 μm (b–f,b’–f’), 3.1 mm (g,h,k,l,o,p), 550 μm (i,j,n,r,t), 1.0 mm (m,q,s), and 2.1 mm (u–w). See also Supplementary Fig. S2.