Abstract
High-throughput behavioral phenotyping is critical to genetic or chemical screening approaches. Zebrafish larvae are amenable to high-throughput behavioral screening because of their rapid development, small size, and conserved vertebrate brain architecture. Existing commercial behavioral phenotyping systems are expensive and not easily modified for new assays. Here, we describe a modular, highly adaptable, and low-cost system. Along with detailed assembly and operation instructions, we provide data acquisition software and a robust, parallel analysis pipeline. We validate our approach by analyzing stimulus response profiles in larval zebrafish, confirming prepulse inhibition phenotypes of two previously isolated mutants, and highlighting best practices for growing larvae prior to behavioral testing. Our new design thus allows rapid construction and streamlined operation of many large-scale behavioral setups with minimal resources and fabrication expertise, with broad applications to other aquatic organisms.
Highlights
High-throughput behavioral tracking offers great potential for large-scale mutant phenotyping (Thyme et al, 2019) and drug screening (MacRae and Peterson, 2015)
Larval zebrafish maintained in 96-well plate format execute diverse behaviors including prepulse inhibition (Burgess and Granato, 2007), sleep (Chiu et al, 2016), seizures (Griffin et al, 2020), prey consumption (Jordi et al, 2018), and responses to visual (Randlett et al, 2019), acoustic (Wolman et al, 2011), or thermal stimuli (Chiu et al, 2016)
Stimulus intensity was inconsistent across setups due to variable construction
Summary
High-throughput behavioral tracking offers great potential for large-scale mutant phenotyping (Thyme et al, 2019) and drug screening (MacRae and Peterson, 2015). Our new design significantly extends systems previously validated in a large-scale mutant screen (Thyme et al, 2019), with more precise control over a broader range of assays and greater ease of construction. This system includes most of the assays of commercially available solutions and accommodates additional modules. We outline best experimental practices for yielding consistent and reliable behavioral data This fully customizable and modular setup can be adapted as new behavioral assays are published, significantly lowering barriers to large-scale phenotyping approaches
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