Figure 5

Figure 5

Tracing of the presynaptic neurons of PCs using the RV method. A solution of pseudotyped RV was injected into the left side of the cerebellum of adult Tg(aldoca:TVA-mCherry); Tg(aldoca:G) fish. The injected fish were reared at 34–35.5°C for 5 days. The brains were then harvested, fixed, and subjected to immunostaining. (A–E) Staining of sagittal sections with anti-mCherry (magenta) and anti-GFP (green) antibodies, and the nuclear marker Hoechst (cyan). (B,C,Ea,Eb) Higher-magnification images of the boxes in (A,D). GFP (Ba,Ca,Ea), TVA-mCherry (Bb,Cb,Eb), and merged images (B,C) are shown. GFP was observed in the molecular layer (ML) and PC layer (PCL; A). GFP was detected in the TVA-mCherry⁺ PCs (B). GFP but not TVA-mCherry was also detected in neurons in the granular layer (GL; C) and the inferior olivary (IOs; E). (F) Schematic summary of the PC afferent tracing results. Scale bar: (A) 100 μm, (B) 20 μm, (C) 20 μm, (D) 200 μm, and (E) 20 μm.