Measuring Larval Zebrafish Behavior: Locomotion, Thigmotaxis, and Startle

Larval zebrafish are genetically tractable, easy to manipulate, and can be generated in large numbers. Their small size also makes them ideal for high-throughput screens for genes or pharmacological compounds that affect behavior. Thus, larval zebrafish are an ideal model system to investigate locomotion and locomotion-based behaviors such as thigmotaxis and startle. In this chapter, I present methods to measure locomotion, thigmotaxis, and response to a startle stimulus in larval zebrafish. These methods use a Zebrabox and Zebralab software (from ViewPoint Life Sciences) in a manner which is suitable for high-throughput analyses. I compare the locomotion and thigmotaxis of several widely available wild-type strains to demonstrate that these protocols are sensitive enough to detect behavioral differences in different genetic backgrounds. I also measure the difference in startle response between AB wild-type and r � 3−/− mutant fish. These protocols will help to standardize analyses of larval zebrafish locomotion and may provide a starting point for behavioral measurement of the large number of zebrafish mutants which have already been identified.