title: A novel high-throughput imaging system for automated analyses of avoidance behavior in zebrafish larvae.
abstract: Early brain development can be influenced by numerous genetic and environmental factors, with long-lasting effects on brain function and behavior. The identification of these factors is facilitated by recent innovations in high-throughput screening. However, large-scale screening in whole organisms remains challenging, in particular when studying changes in brain function or behavior in vertebrate model systems. In this study, we present a novel imaging system for high-throughput analyses of behavior in zebrafish larvae. The three-camera system can image 12 multiwell plates simultaneously and is unique in its ability to provide local visual stimuli in the wells of a multiwell plate. The acquired images are converted into a series of coordinates, which characterize the location and orientation of the larvae. The developed imaging techniques were tested by measuring avoidance behaviors in seven-day-old zebrafish larvae. The system effectively quantified larval avoidance and revealed an increased edge preference in response to a blue or red 'bouncing ball' stimulus. Larvae also avoid a bouncing ball stimulus when it is counter-balanced with a stationary ball, but do not avoid blinking balls counter-balanced with a stationary ball. These results indicate that the seven-day-old larvae respond specifically to movement, rather than color, size, or local changes in light intensity. The imaging system and assays for measuring avoidance behavior may be used to screen for genetic and environmental factors that cause developmental brain disorders and for novel drugs that could prevent or treat these disorders.
authors.affiliation: Department of Molecular Biology, Cell Biology and Biochemistry, Brown University, Providence, RI, USA.
authors.forename: Sean D
authors.initials: SD
authors.lastname: Pelkowski
authors.forename: Mrinal
authors.initials: M
authors.lastname: Kapoor
authors.forename: Holly A
authors.initials: HA
authors.lastname: Richendrfer
authors.forename: Xingyue
authors.initials: X
authors.lastname: Wang
authors.forename: Ruth M
authors.initials: RM
authors.lastname: Colwill
authors.forename: Robbert
authors.initials: R
authors.lastname: Creton
mesh_terms: D000818:Animals; D001362:Avoidance Learning; D057166:High-Throughput Screening Assays / Q000295:instrumentation* / Q000379:methods*; D007091:Image Processing, Computer-Assisted / Q000295:instrumentation* / Q000379:methods*; D007814:Larva; D010775:Photic Stimulation / Q000379:methods*; D014741:Video Recording / Q000379:methods*; D015027:Zebrafish