Genetic Identification of Neural Circuits Essential for Active Avoidance Fear Conditioning in Adult Zebrafish

Inhibition or ablation of neuronal activity combined with behavioral assessment is crucial in identifying neural circuits or populations essential for specific behaviors and to understand brain function. In the model vertebrate zebrafish, the development of genetic methods has allowed not only visualization but also targeted manipulation of neuronal activity, and quantitative behavioral assays allow precise measurement of animal behavior. Here, we describe a method to inhibit a specific neuronal population in adult zebrafish brain and assess their role in a learning behavior. We employed the Gal4–UAS system, gene trap and enhancer trap methods, and isolated transgenic zebrafish lines expressing Gal4FF transactivator in specific populations of neurons in the adult zebrafish brain. In these lines, a genetically engineered neurotoxin, botulinum toxin B light chain, was expressed and the fish were assessed in the active avoidance fear conditioning paradigm. The transgenic lines that showed impaired avoidance response were isolated and, in these fish, the Gal4-expressing neurons were analyzed to identify the neuronal circuits involved in avoidance learning.