Figure 3.

AAV9-aGRIK2 treatment decreases seizure in epileptic mice and inter-ictal epileptiform discharges in human organotypic hippocampal slices

(A) Timeline of the experimental procedure in epileptic mice (mTLE, top) injected bilaterally in the hippocampus with either AAV9-control (5.0E+9 gc/hippocampus) or AAV9-GRIK2 (at different doses) and example of a typical electrographic seizure (ictal event, bottom). (B) Number of seizures per day in the presence of either AAV9-control or AAV9-GRIK2. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗∗p < 0.0001, one-way ANOVA followed by Tukey’s multiple comparisons test. (C) Photomicrographs showing a representative example of dentate gyrus granule cells (DGCs) transduced with AAV9-GFP (AAV9.hSyn1.GFP) in hippocampal organotypic slices from a patient with temporal lobe epilepsy (TLE), using double immunostaining with anti-Prox1 (left), and anti-GFP (middle) antibodies; Molecular layer (ML), hilus (H), scale bars, top, 50 μm, middle, 100 μm, and bottom, 200 μm. (D) Quantification of miR1 expression level. (E) Quantification of mi2R expression level. (F) Quantification of GRIK2 mRNA in tissue treated with AAV9-aGRIK2 normalized to treatment with the AAV9-control. Student’s t test, ∗∗p < 0.01. (G) Representative traces of LFPs recorded in human hippocampal organotypic slices transduced with either control AAV9-control (top) or AAV9-aGRIK2 (bottom, 2.0E+10 gc per slices); enlarged interictal-like epileptiform discharges (IEDs) shown as upper and lower traces (scale bar, 50 ms); note the decrease of IEDs in slices treated with AAV9-aGRIK2. (H) Quantification of the frequency of IEDs in human organotypic hippocampal slices from five patients with mTLE treated with either AAV9-control or AAV9-aGRIK2; control and treated slices from the same patient are paired (lines) for Student’s t test, ∗p < 0.05. Data are reported as mean ± SEM.