Larval Zebrafish Locomotor Assay

At 10 dpf, morphologically normal larvae were removed from flasks using a sterile transfer pipet and placed into 48-well plates containing 500 µl of FS-10% HBSS per well. Plates were sealed with Microseal A film (BioRad, #MSA5001), wrapped in Parafilm, and placed in the dark in a temperature controlled behavior testing room at 26 °C, for at least 2 hr prior to testing. For testing, microtiter plates were placed on a Noldus tracking apparatus and locomotor activity was recorded for 40 mins. The light program consisted of a 20-min acclimation period in the dark (0 lux) followed by a 10 min light period (5 lux) and a 10 min dark period (0 lux). All tests were carried out between 11:15 am–4:00 pm. Videos were analyzed using Ethovision software Version 12 (Noldus Information Technology) as described previously^{48 (link)}.