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Fig. 1 | Frontiers in Zoology

Fig. 1

From: Enteric neuroanatomy and smooth muscle activity in the western diamondback rattlesnake (Crotalus atrox)

Fig. 1

Representative immunofluorescent images of different regions of the GI tract of Crotalus atrox. A shows a transverse section of the stomach, stained in green for PGP 9.5. Nerve cells of the submucous (SMP) and myenteric plexus (MP) are marked with red and blue arrowheads, respectively. The orange and yellow arrows show neuronal elements innervating the mucosa and the circular muscle layer, respectively. MUC: mucosa, MM: muscularis mucosae, CM: circular muscle, LM: longitudinal muscle. B, C illustrate the difference between the submucous (SMP; B) and myenteric plexus (MP; C) of the colon. In the SMP, only one individual neuronal cell body (green, stained for Hu, the pan-neuronal marker; marked with a white arrow) is seen with connecting fiber tracts (red, stained for VIP), while in the MP larger ganglia with several nerve cell bodies (green) are visible. D Image of the oesophagus MP demonstrates that ChAT (red) and NOS (green) rarely colocalize and ChAT and NOS positive neurons are generally grouped together (ChAT: blue circle; NOS: white circle). E–G show examples of VIP-stained nerve cells in different regions and plexuses. In panel E, two neuronal cell bodies (white arrows) of the proximal small intestine SMP are visible, which are both stained for NOS (green) and VIP (red). In panel F, in the proximal stomach SMP, one neuronal cell body (white arrow) stained for NOS, but not for VIP, and another neuronal cell body (blue arrow) stained for VIP, but not for NOS are present. G shows a different colour code in the MP of the distal small intestine. Three neuronal cell bodies are ChAT positive (green, marked with a white arrow), while no cell bodies are stained for VIP (red), only fiber tracts and varicous endings. H–K presents the same ganglion in the MP of the colon. Panel H shows the staining for Hu in blue, which marked all neuronal cell bodies, but no nerve fibers. In panel I, several neuronal cell bodies (marked with white arrows) are stained for TH (green), corresponding to dopaminergic neurons. In panel J, no cell bodies, only fibers are stained with DBH (red), a marker of noradrenalin synthesis. K shows the merged image of all 3 stainings. Note that apart from blue, most neuronal cell bodies are also stained for TH with green (white arrows), but three neuronal cell bodies (orange arrows) appear in pure blue colour only, representing TH-negative nerve cells

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