title: Use of the TetON System to Study Molecular Mechanisms of Zebrafish Regeneration.
abstract: The zebrafish has become a very important model organism for studying vertebrate development, physiology, disease, and tissue regeneration. A thorough understanding of the molecular and cellular mechanisms involved requires experimental tools that allow for inducible, tissue-specific manipulation of gene expression or signaling pathways. Therefore, we and others have recently adapted the TetON system for use in zebrafish. The TetON system facilitates temporally and spatially-controlled gene expression and we have recently used this tool to probe for tissue-specific functions of Wnt/beta-catenin signaling during zebrafish tail fin regeneration. Here we describe the workflow for using the TetON system to achieve inducible, tissue-specific gene expression in the adult regenerating zebrafish tail fin. This includes the generation of stable transgenic TetActivator and TetResponder lines, transgene induction and techniques for verification of tissue-specific gene expression in the fin regenerate. Thus, this protocol serves as blueprint for setting up a functional TetON system in zebrafish and its subsequent use, in particular for studying fin regeneration.
authors.affiliation: Institute for Biochemistry and Molecular Biology, Ulm University.
authors.forename: Daniel
authors.initials: D
authors.lastname: Wehner
authors.affiliation: Institute for Biochemistry and Molecular Biology, Ulm University.
authors.forename: Christopher
authors.initials: C
authors.lastname: Jahn
authors.affiliation: Institute for Biochemistry and Molecular Biology, Ulm University; gilbert.weidinger@uni-ulm.de.
authors.forename: Gilbert
authors.initials: G
authors.lastname: Weidinger
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D015870:Gene Expression; D012038:Regeneration / Q000235:genetics / Q000502:physiology*; D013623:Tail / Q000502:physiology; D013752:Tetracycline / Q000494:pharmacology*; D015533:Transcriptional Activation; D015027:Zebrafish / Q000235:genetics / Q000502:physiology*
chemical_list: D013752:Tetracycline