Visualizing Spatial Expression Pattern of Anoctamin-1 (Ano1) in the Zebrafish Gastrointestinal Tract

Anoctamin-1 (Ano1) is expressed in mammalian interstitial cells of Cajal (ICC) and encodes a Ca2+-activated Cl- channel that may be functionally important for gastrointestinal (GI) smooth muscle pace making. Reduced Ano1 expression in zebrafish larvae is associated with delayed GI transit and uncoordinated GI motility patterns. Objective: To visualize the spatial expression pattern for Ano1 in the zebrafish GI tract using in situ hybridization. Methods: Using in situ hybridization, ano1 mRNA expression will be visualized in adult zebrafish tissue. Ano1 protein expression will also be shown visually using immunofluorescence. Gross tissue morphology will be confirmed using hematoxylin and eosin staining. Anticipated Results: Ano1 expression in ICC has been shown in mammalian tissues and similar Ano1 expression patterns in zebrafish ICC is anticipated. Expression of Ano1 mRNA will be determined using in situ hybridization and protein expression will be determined using immunohistochemistry. Ano1-positive ICC in the myenteric plexus region will be consistent with the role of Ano1 in ICC pacemaker function. Summary/Future Direction: Confirmation of Ano1 expression patterns in zebrafish GI tissue is the first step towards understanding its functional role. In the future, a spatiotemporal study will be performed to understand when and where Ano1 expression occurs during embryonic development. Supported by NIH DK0715880