Fig. 6: Optogenetic-driven astrocytic Na⁺-K⁺-ATPase rescues seizure susceptibility of FCD rats.

a Establishment of FCD model in SD rats and timeline design for behavioral experiments. b Representative immunostaining images showed NeuN⁺ cells in nondysplastic (control) and dysplastic (FCD) cortex. Scale bar, 50 μm and 25 μm. c Representative cortical EEG traces recorded from dysplastic FCD rat showed several forms of spontaneous epileptic waves. d Representative energy spectra from control and FCD rats injected with PTZ. e–h Seizure susceptibility of control and FCD rats was tested after PTZ injection (i.p. injection, 60 mg/kg) and seizure stage (e), EEG onset (f), latency to GS (g) and GS duration (h) were recorded. i Fluorescent images showed the expression pattern of Na⁺-K⁺-ATPase protein (ATP1A2) in control and FCD cortical tissue. j Representative western blot bands and quantification of Na⁺-K⁺-ATPase protein (ATP1A2) expression in control and FCD cortex. GAPDH was used as the internal control. k FCD rats were injected with ChR2-EYFP and/or pAAV-shortGFAP-MCS-EGFP-3FLAG-mir30shRNA(Atp1α2) in the M1 and implanted with fiber and electrode for blue light stimulation and EEG recording. l Fluorescent images showed shRNA(Atp1α2) co-localization with GFAP⁺ astrocytes and knockdown of astrocytic Na⁺-K⁺-ATPase protein (ATP1A2) expression in FCD cortical tissue. Scale bar, 20 μm. m–p Effects of optogenetic stimulation of astrocytes on seizure stage (m), EEG onset (n), latency to GS (o) and GS duration (p) in FCD-ChR2^M1rats, in the condition of astrocytic Na⁺-K⁺-ATPase inhibition by shRNA-knockdown. *p < 0.05, **p < 0.01, ***p < 0.001 compared with each control group. Data shown as mean ± s.e.m. The number of mice used is indicated in figures. For detailed statistical information, see Supplementary Data 1. Uncropped version of western blots that labeled with the relevant panel and antibody were provided in Source data western blot. Source data are provided as a Source Data file.