Figure 3

Figure 3

Evolution of the excitatory-inhibitory neuronal population in the scn1Lab model. (A_1-3,B_1-3) Dorsal view of 3 dpf Tg[Gad1b:GFP; Vglut2a:DsRed] control (A_1-3) (N = 8) and scn1Lab (B_1-3) (N = 8) living larvae. (A₁,B₁) Inhibitory neurons (Gad1b:GFP) in 3 dpf control (A₁) and scn1Lab (B₁) larvae. (A₂,B₂) Excitatory neurons (Vglut2a:DsRed) in 3 dpf control (A₂) and scn1Lab (B₂) larvae. (A₃,B₃) Merged image of 3 dpf control (A₃) and scn1Lab (B₃) larvae. (C) Dorsal view schematic of a 4 dpf larva. The red rectangle shows the region of interest. (E_1-3,F_1-3) Dorsal view of 4 dpf Tg[Gad1b:GFP; Vglut2a:DsRed] control (E_1-3) (N = 8) and scn1Lab (F_1-3) (N = 8) living larvae. (E₁,F₁) Inhibitory neurons (Gad1b:GFP) in 4 dpf control (E₁) and scn1Lab (F₁) larvae. (E₂,F₂) Excitatory neurons (Vglut2a:DsRed) in 4 dpf control (E₂) and scn1Lab (F₂) larvae. (E₃,F₃) Merged image of 4 dpf control (E₃) and scn1Lab (F₃) larvae. (H_1-3,I_1-3) Dorsal view of 5 dpf Tg[Gad1b:GFP; Vglut2a:DsRed] control (H_1-3) (N = 8) and scn1Lab (I_1-3) (N = 8) living larvae. (H₁,I₁) Inhibitory neurons (Gad1b:GFP) in 5 dpf control (H₁) and scn1Lab (I₁) larvae. (H₂,I₂) Excitatory neurons (Vglut2a:DsRed) in 5 dpf control (H₂) and scn1Lab (I₂) larvae. (H₃,I₃) Merged image of 5 dpf control (H₃) and scn1Lab (I₃) larvae. (D,G,J) Number of neurons in control and scn1Lab larvae at 3 dpf (D), 4 dpf (G) and 5 dpf (J). (K,L,M) Number of inhibitory and excitatory neurons in control and scn1Lab larvae at 3 dpf (K), 4 dpf (L) and 5 dpf (M). All images are representative 20 µm stacks acquired using LSM 880 Zeiss laser scanning confocal microscope equipped with a WPApo 20x/1.0 objective. Scale bar 40 µm. N = number of embryos. Error bars on all graphs represent standard error mean (SEM). * p < 0.05; ** p < 0.01; ***,p < 0.001 indicate a statistically significant difference between control and scn1Lab larvae. Statistically significant difference between inhibitory and excitatory neuronal population is indicated as $$, p < 0.01; $$$, p < 0.001. p-values were determined using Student’s unpaired t-test.