Figure 4.

Validation of identified marker genes with double fluorescent in situ hybridization. In each part, an in situ hybridization image of a coronal MVN section is shown in the top row (i), and magnified images of the white-boxed area are shown in the bottom row (ii–iv). A, Slc17a7 (magenta) and Crh (inset, green). The in situ hybridization signals of Slc17a7 and Crh were sparsely distributed over the MVN, and greatly overlapped each other. The arrowhead indicates the granule cell layer in the cerebellum. B, Adcyap1 (magenta) and Slc17a6 (green). Adcyap1-positive cells were scattered in the MVN, especially near the fourth ventricle. In the majority of cases, they colabeled with Slc17a6. C, Coch (magenta) and Gad1 (green). Coch-positive cells were nearly always colabeled by Gad1. The hybridization signal was also observed in the choroid plexus in the fourth ventricle. D, Crhbp (magenta) and Gad1 (green). Crhbp-positive cells were also colabeled by Gad1 (green, iii and iv), forming a dense cluster near the fourth ventricle and showing distinct distribution from that of Coch-positive cells. Scale bar: 150 μm for the top row images (i) and the inset images in A; 50 μm for the bottom images (ii, iii, iv).