Automated, high-throughput, in vivo analysis of visual function using the zebrafish

Abstract

Background

Modern genomics has enabled the identification of an unprecedented number of genetic variants, which in many cases are extremely rare, associated with blinding disorders. A significant challenge will be determining the pathophysiology of each new variant. The zebrafish is an excellent model for the study of inherited diseases of the eye. By 5 days-post-fertilization (dpf) they have quantifiable behavioral responses to visual stimuli. However, visual behavior assays can take several hours to perform or can only be assessed one fish at a time.

Results

To increase the throughput for vision assays, we used the Viewpoint Zebrabox to automate the visual startle response and created software, Visual Interrogation of Zebrafish Manipulations (VIZN), to automate data analysis. This process allows 96 zebrafish larvae to be tested and resultant data to be analyzed in under 35 minutes. We validated this system by disrupting function of a gene necessary for photoreceptor differentiation and observing decreased response to visual stimuli.

Conclusions

This automated method along with VIZN allows rapid, high-throughput, in vivo testing of zebrafish’s ability to respond to light/dark stimuli. This allows the rapid analysis of novel genes involved in visual function by morpholino, CRISPRS, or small molecule drug screens.