Figure 8. Validation of SNAP-25 as a NENs marker.

(a) SNAP-25 expression is restricted to the neural crest lineage in the adult myenteric ganglia. SNAP-25 expression (green) co-localizes with tdTomato (red) but not with the MENs marker MHCst (cyan) as observed in 2D and orthogonal views of a myenteric ganglia from a Wnt1-Cre:Rosa26 ^lsl-tdTomato mouse that was immunolabeled with antibodies against MHCst and SNAP-25. Nuclei were labeled with DAPI (blue). Scale bar denotes 10 µm. (b) In the adult male Snap25^GcaMP knock-in mice, the expression of Snap25-GCaMP/GFP (green) is restricted to a subset of Hu-expressing (red) myenteric neurons in the adult murine small intestinal myenteric ganglia (yellow arrows), while many neurons (red arrows) do not show any detectable expression of Snap25. Nuclei were labeled with DAPI (blue). Scale bar denotes 10 µm. (c) Orthogonal views of z-stack of an image of the myenteric ganglion from the small intestinal LM-MP tissue from an adult male Snap25^GcaMP knock-in mice shows that the expression of Snap25-GCaMP/GFP (green) is exclusive of the expression of the MENs marker MHCst (red). Nuclei were labeled with DAPI (blue). Scale bar denotes 10 µm.