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Fig. S3
Effect of ACh-Cl on the AChE activity and ENS neuron development.
(a) Immunohistochemical staining of HuC/D in both control and ACh-Cl treated fish at 6 dpf. The data show no significant difference between the control and chemical application groups. Left pannels show the whole body staining pattern of HuC/D whereas middle and right represent the high magnifications of the areas that boxed in left panels. Right panels are the images of middle merged with DIC. (b) The AChE activity detection assay reveals that the AChE activity does not show obvious difference (middle panels) after transit application of ACh-Cl but reduces obviously after ACh-Cl treatment for 12 hours (right panels) compared with control (left panels) in the gut but not elsewhere in the body. Upper panels show the whole body staining pattern of AChE whereas bottom panels represent the high magnifications of the boxed images correspondingly. The red arrows indicate the intestinal bulb region where the peristalsis is identified.