Nunc 055431 150687

The paradigm used in this set of experiments constituted a set of 5 flashes (1 s of light flash—49 s of darkness, repeated 5 times), performed in order to trigger epileptic behavior. For these experiments, 48-well plates (Nunc 055431/150687, Thermo Scientific, Waltham, MA, USA) were also used and placed in the chamber in line with the camera (one larva per well). To allow the habituation of the larvae to the plate, they were plated at least 3 h before the behavioral recording and all the tests had a 10 min light acclimatation phase inside the recording chamber. The same plate distributions as in the locomotion activity assay of the selected mutants were used.

To evaluate possible locomotion alteration in the generated crispants, the standardized 25 min dark/light cycles paradigm (5 min darkness—5 min light up to 25 min) was used. The total distance moved (mm) was analyzed. For these experiments, 48-well plates (Nunc 055431/150687, Thermo Scientific, Waltham, MA, USA) were used and placed in the chamber in line with the camera (one larva per well). To allow the habituation of the larvae to the plate, they were plated at least 3 h before the behavioral recording and all the tests had a 10 min light acclimatation phase inside the recording chamber. As aforementioned, different plate distributions were used, allowing to always have a scrambled control on each plate.