Double in situ hybridization of VAChT and VGAT.Whole mounts with anterior to the left and whole mount scale bars are 50 μm. A-D: Serial confocal fluorescence images of a 18-hr neurula. A: VAChT-positive neurons (green). B: VGAT-positive neurons (red). C: Merger of A and B. D: Magnification of the anterior neural tube shown in C. E,F: Lateral views of a 22-hr neurula. In E, the dark purple and red staining respectively show VAChT and VGAT expression. In F, the superimposition of brightfield and epifluorescence shows both VAChT-positive and VGAT-positive neurons.