Fig. 7
From: Neuregulin 1/ErbB4 signaling contributes to the anti-epileptic effects of the ketogenic diet
ErbB4 deletion in PV+ interneurons is necessary for the anti-seizure effect of the KD. a Mice breeding paradigm. ErbB4f/f mice were crossed with PV::Cre mice; the resulting PV::Cre;ErbB4f/+ mice were crossed with ErbB4f/+ mice to generate PV::Cre;ErbB4 (PV-KO) and ErbB4f/f (control) mice. b Reduced ErbB4 expression in the hippocampus of PV-KO mice. Hippocampal lysates of 2-month-old mice were probed for ErbB4, and GAPDH was taken as a loading control. Shown were representative blots of each genotype. c Quantitative analysis of data in (b). n = 6 mice per group. Student’s t test, t(10) = 4.853, P = 0.0007. d Diagram of hippocampal slice recording. Pyramidal neurons in the CA1 region were clamped in whole-cell configuration. Evoked postsynaptic currents were recorded under stimulation by using a concentric bipolar electrode. e Representative eIPSC traces. Scale bars, 100 ms and 1000 pA. f Deletion of ErbB4 in PV+ interneurons diminished the effect of KD on eIPSC amplitudes. n = 16 neurons from 3 mice of “Control + CD” group; n = 16 neurons from 3 mice of “Control + KD” group; n = 16 neurons from 3 mice of “PV-KO + CD” group; n = 14 neurons from 3 mice of “PV-KO + KD” group. Two-way ANOVA, “Control + CD” vs “Control + KD”: F(1,269) = 57.97, P < 0.0001; “PV-KO + CD” vs “PV-KO + KD”: F(1,252) = 3.158, P = 0.0768. g Representative sIPSC traces. Scale bars, 2 s and 50 pA. h Deletion of ErbB4 in PV+ interneurons diminished the effect of KD on sIPSC frequency. n = 16 neurons from 3 mice of “Control + CD” group; n = 16 neurons from 3 mice of “Control + KD” group; n = 16 neurons from 3 mice of “PV-KO + CD” group; n = 14 neurons from 3 mice of “PV-KO + KD” group. Student’s t test, “Control + CD” vs “Control + KD”: t(30) = 4.985, P < 0.0001; “PV-KO + KD” vs “PV-KO + KD”: t(28) = 1.873, P = 0.0715. i Schematic of experimental design. Control and PV-KO mice that were feed with CD or KD for 3 weeks were subject to KA infusion to induce seizures. j The development of seizure with time following KA infusion. n = 7 mice for each group. Repeated two-way ANOVA, “Control + CD” vs “Control + KD”: F(1,12) = 5.685, P = 0.0345; “PV-KO + CD” vs “PV-KO + KD”: F(1,12) = 1.311, P = 0.2746. k ErbB4 deletion in PV+ interneurons diminished the effect of KD on the averaged seizure score. n = 7 mice for each group. Student’s t test, “Control + CD” vs “Control + KD”: t(12) = 2.384, P = 0.0345; “PV-KO + CD” vs “PV-KO + KD”: t(12) = 1.145, P = 0.2746