Utilizing Behavioral and Molecular Techniques to Study Gap Junction Channels in Developing Zebrafish
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Date
2020
Authors
Reich, Laura
Journal Title
Journal ISSN
Volume Title
Publisher
University of Oregon
Abstract
Animal behavior requires coordination between the nervous and muscular systems. These systems communicate at specialized subcellular structures, found within and between systems, that allow the cells to coordinate their activity to achieve movement. One type of communication arises from gap junction channels (GJCs), which are built by the Connexin (Cx) family of proteins that allow for direct ionic and small molecule exchange between interconnected cells. The GJC family is large with up to 20 individual genes encoded in the human genome. Given this complexity, it is unknown how individual Cxs contribute to behavior. We are using the embryonic zebrafish to address this question due to its rapid development, genetic access, and its first behavior, spontaneous coiling, which requires GJCs. We first identified Cxs that were likely to contribute to coiling using a combination of RNA-seq and RNA detection in vivo and found a previously uncharacterized Cx, Cx46.8, expressed in slow muscle fibers. To understand Cx46.8’s involvement in coiling, we developed an automated behavioral tracking system, using DeepLabCuts (DLC), to track movement during spontaneous coiling. Using this system, we found that animals lacking Cx46.8 have defects in coiling, indicating that we have identified a novel Cx that contributes to behavior. Further experimentation will utilize DLC, in addition to molecular techniques, to unravel the molecular and functional mechanisms of Cx46.8 and other Cxs that contribute to GJC communication in behavior.
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Keywords
biology, zebrafish, gap junction channel, connexin, DeepLabCut