Behavioral screening of gal4-enhancer trap lines with UAS-tetanus toxin in adult zebrafish

Akira Muto, Maximiliano Suster, Kazuhide Asakawa, and Koichi Kawakami

Division of Molecular and Developmental Biology, National Institute of Genetics (NIG), Japan

  Adult zebrafish (Danio rerio) show a repertoire of behaviors such as locomotion, shoaling, and feeding. These behaviors are genetically coded in part and also can be learned and/or modulated by environmental factors. Our objective is to study cellular mechanisms of animal behaviors using zebrafish as a vertebrate animal model. To identify subset of neurons that are responsible for fish behaviors, we used neurotoxins to silence neurons using gal4-UAS system. We generated a large collection of zebrafish lines that expressed Gal4 driver in a specific manner at larval stage, and selected some lines based on their UAS-GFP expression in the adult brain. We also developed transgenic effector fish carrying the tetanus toxin light chain (TeTxLC) gene downstream of UAS. We crossed the Gal4 fish with the UAS-TeTxLC and screened double transgenic fish for a behavioral phenotype in adult. For the screening, we developed efficient behavioral screening assays to assess locomotor activity, social interaction, and learning and memory. Place preference test in the presence of a group of fish in one end of a test tank was used to assess tendency for social interaction in the test fish. Operant behavior with feeding activity was used for learning and memory assay. Our experimental set up offers a powerful means for discovery of cellular mechanism that links genes and behaviors in an unbiased manner. Our results will shed light on how animal behaviors are determined in a context with gene-environment interaction.