Figure 4. Normal intrinsic firing properties during epileptogenesis but increased mTOR-dependent soma size increase in SSCx L2/L3 pyramidal neurons.

(A) Timeline of the patch-clamp experiments. The colored squares indicate the day of patch-clamp measurements. (B and C) Average number of action potentials of SSCx L2/L3 pyramidal neurons after increasing current injections of Tsc1-Cre^– and Tsc1-Cre⁺ mice measured on day 8 (B: Tsc1-Cre^– n = 16, 4 mice; Tsc1-Cre⁺ n = 19, 5 mice) and day 12 after gene deletion (C: Tsc1-Cre^– n = 22, 4 mice; Tsc1-Cre⁺ n = 23, 3 mice; 2-way RM ANOVA; Dunnett’s with Tsc1-Cre^– as control). (D) Experimental timeline of the patch-clamp experiments and fluorescence staining for NeuN (green) and DAPI (blue). The squares indicate the day of patch-clamp measurements. (E and F) The average number of action potentials of L2/L3 pyramidal neurons in the SSCx after increasing current injections of Tsc1-Cre^– and Tsc1-Cre⁺ mice measured on day 12 with rapamycin treatment starting on day 4 (Tsc1-Cre^– n = 13, 4 mice; Tsc1-Cre⁺-Rap 4 n = 11, 4 mice) and day 8 (Tsc1-Cre⁺-Rap 8 n = 15, 4 mice; 2-way RM ANOVA; Dunnett’s with Tsc1-Cre^– as control). (G) Representative confocal pictures with a NeuN (green) and DAPI (blue) staining of SSCx L2/L3 pyramidal neurons of Tsc1-Cre^–, Tsc1-Cre⁺ day 12, Tsc1-Cre^–-Rap 4, and Tsc1-Cre⁺-Rap 8 mice. Scale bar demonstrates 50 μm. (H) Average soma size SSCx L2/L3 pyramidal neurons per slice (15–20 cells per slice) per group (black: Tsc1-Cre^–: n = 24, 5 mice; red: Tsc1-Cre⁺ day 12: n = 11, 4 mice; blue: Tsc1-Cre⁺-Rap 4: n = 19, 3 mice, and green: Tsc1-Cre⁺-Rap 8: n = 18, 4 mice; 1-way ANOVA; Dunnett’s with Tsc1-Cre^– as control). Error bars indicate SEM. ***P < 0.001, ****P < 0.0001.