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Fig. 1 Experimental design of the tube assay and overview of zebrafish training. (A) Schematic diagram of the experimental setup with dimensions of the L-shaped tube indicated. Note that for the internal diameter, two values are given to correspond with the diameter of the tube used first (larger) and the subsequently used tube (smaller diameter). A single zebrafish was introduced into the vertical short arm of the tube. Placement of the tube inside the tank is shown (B) Time to escape from the tube, that is, length of time the fish took to traverse the distance from point A to point B (exit point) (s, seconds) is shown as a function of trials in the 70 cm long and 2.1 cm inner diameter tube. Results for two individual zebrafish, Fish #1 and Fish #2, are shown. (C) Average escape time for wild-type zebrafish over ten trials in the 70 cm tube with 1.6 cm diameter (n = 17). (D) Schematic representation of the optimized paradigm indicating the habituation and training trials.