Ethovision xt 14 15

All compounds were diluted in 100% methanol (MeOH) and experiments were performed in a HEPES buffered E3 (HE3) medium (5 mM NaCl, 0.17 mM KCl, 0.33 mM CaCl2-2H2O, 0.33 mM MgSO4-7H2O, 10 mM HEPES, pH 7.2). Individual 120 h post fertilization (hpf) larval zebrafish were transferred to a 48-well microtiter plate in 500 μL of HE3 media. Larvae were acclimated for at least 1 hour in a lighted 28.5°C incubator (photon flux: 3–5 μmol s−1 m²) prior to experimentation and larval behavior was analyzed using DanioVision larval tracking systems with EthoVisionXT14/15 software (Noldus Information Technology Inc., Virginia, USA). Distance traveled was measured using dynamic subtraction at 28.5°C over 120 min with the first 90 min under lighted conditions (15 µmol m-2 s-1), followed by alternating 5-min dark/light cycles. Each larvae represents an independent measurement. Any larvae that were dead or displayed phenotypic abnormalities were removed from analysis. 12 larvae were used in each experimental condition, and at least 2 replicates of each concentration were performed.

The OFT is a commonly used experiment to measure the general locomotor activity, novel environment exploration, and anxiety-like behavior¹¹ . Mice were placed in an acrylic-made open square arena (48 × 48 cm) and allowed to explore for 10 minutes freely. The video was recorded by the ceiling mounted camera and analyzed by Ethovision XT 14/15 (Noldus Information Technology), including the distance moved and duration of stay. The center of the arena was defined as the center zone (24 × 24 cm) and the peripheral zone was defined as 6 cm inward from the chamber’s boundary.

The EPM is a test to assess anxiety-like behavior and innate fear¹¹⁵ . This maze is composed of a “+“ shape with two opposite directions of opened, closed arms and a center area (5 × 5 cm). Both opened arms and closed arms are 30 cm in length and 5 cm in width, but only the closed arms had the 15 cm heighten walls around. The whole maze was set up 55 cm above the floor to induce the propensity of mice acrophobia. Mice were allowed to freely explore the maze for 5 min. The video was recorded and analyzed the distance moved, frequency of entry, and duration in opened and closed arms by Ethovision XT 14/15 (Noldus).