Fig. 2

depdc5 Knockout Results in Increased mTOR Activity, Hypoactivity, Epileptiform Discharges, and Seizure Susceptibility

(A) Western blotting of 7-dpf larval lysates to examine phospho-S6 levels in depdc5^+/+, depdc5^+/−, and depdc5^−/− larvae showing hyperactivation of mTOR signaling. n = 3; p(WT vs HT) = 0.0012; p(WT vs HM) = 0.0455.

(B) Images of 6-dpf depdc5^+/+ and depdc5^−/− larval brains (transversal sections across the optic tectum) immunostained with phospho-S6 antibodies and counterstained with DAPI. The panels on the extreme right depict 3D reconstruction of single cells in depdc5^+/+ and ^−/− brains. Scale bars from left to right: 20 μm, 2.5 μm, and 1.5 μm.

(C) Spontaneous extracellular electrographic activity recorded from the optic tectum of depdc5^+/+ or depdc5^−/− larvae at 9 dpf showing representative epileptiform activity of depdc5^−/− larvae displaying polyspiking discharges.

(D) Percentage of depdc5^+/+ and depdc5^−/− larvae showing epileptiform activity in the brain. N = 4, n = 10/genotype; Student’s t test p(WT vs HM) < 0.0001.

(E) Distance swam by 8-dpf larvae across two 2-hr dark/4-hr light cycles.

(F) Spontaneous coiling activity in 20-hpf embryos monitored over 20 min represented as the percentage of total time during which the embryos were active. N = 3, n = 20; Student’s t test p(WT versus HM) < 0.0001.

(G) Distance swam by 2-dpf embryos upon exposure to 3 mM PTZ represented as fold change compared to basal distance swam by wild-type larvae at t₀. N = 3, n > 10; Student’s t test, ^∗p < 0.05.

(H) Representative swimming tracks of 2-dpf embryos during a 10-min period following a 3 mM PTZ exposure (tracks of 3 replicates are shown in rows for each genotype).