Abstract
The objective of this protocol is to provide a detailed description for the construction and use of a behavioral apparatus, the zBox, for high-throughput behavioral measurements in larval zebrafish (Danio rerio). The zBox is used to measure behavior in multiple individuals simultaneously. Individual fish are housed in wells of multi-well plates and receive acoustic/vibration stimuli with simultaneous recording of behavior. Automated analysis of behavioral movies is performed with MATLAB scripts. This protocol was adapted from two of our previously published papers (Levitz et al., 2013; Pantoja et al., 2016). The zBox provides an easy to setup flexible platform for behavioral experiments in zebrafish larvae.
Keywords: Zebrafish, Behavior, Individuality, Neuromodulation, High-throughput, Startle, Larvae
Background
Behavioral differences between individuals in a population are ubiquitous and likely play a role in adaptation to varying selective pressures during evolution. However, variation in behavior among individuals is often ignored when the quantitation of the behavior of groups is presented as means and associated dispersions. In this protocol, we describe an approach to characterize individuality in the habituation of the acoustic startle behavior at the population level in zebrafish (Danio rerio) larvae. Our approach and set-up can be easily adapted to studies of individuality in other zebrafish larval behaviors.
Materials and Reagents
Equipment
Software
Procedure
Data analysis
Recipes
Acknowledgments
This work supported by the National Institutes of Health Nanomedicine Development Center for the Optical Control of Biological Function (PN2EY018241) and the Human Frontier Science Program (RGP0013/2010). The zBox was initially developed by Dr. David Schoppik in Dr. Alex Schier’s laboratory. The work and experimental approach described here have been adapted from previous research published in Nature Neuroscience (Levitz et al., 2013) and Neuron (Pantoja et al., 2016). Further information about uses for the zBox are reported in a study that investigated the role of neuropeptides in the partition of arousal behaviors in zebrafish (Woods et al., 2014). In addition, work by Zhou et al. can be used as a complementary set of methods for the utilization of this apparatus (Zhou et al., 2014).
References
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