Experimental overview. (A) Schematic timeline of experimental interventions. Adult zebrafish were trained twice weekly for two weeks (black bars), with their final training session recorded (black arrow). After a three-week rest period, the fish were randomly allocated to the control and experimental groups. OKA treatment lasted nine consecutive days. PBM treatments were applied on days 7, 8, and 9 (orange bars). Behavioral tests were conducted on Day 10 (blue bar). (B) Schematic setup of the PBM treatment. An anesthetized zebrafish was positioned under a stabilized cardboard with an aperture of 2 mm by 3 mm to target the brain with a laser beam. Depending on the group, PBM at 3 or 4.5 J/cm2 was applied for three days in a row. The untreated controls underwent the same procedure, but the laser was off. (C) Schematic of the T-maze showing its dimensions: start chamber (S) where fish were held for initial adaptation, central arm (C), left arm (L), and right reward arm (R). (D) Diagram of the T-maze showing color-coded zones used by the TrackAnalyzer module to quantify time spent and distance covered in each zone. The right reward arm (black) was associated with food. (E) Two representative tracking plots recorded at the end of the learning/training phase (three weeks before drug treatment) display typical zebrafish behavior. (F) Quantification of all recorded plots showing the percentage of time spent and distance covered in each zone. The bar graphs show means ± s.e.m while open circles display individual data points. The tracking plots of ten randomly selected fish were analyzed. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.
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