Fig. 5: Overexpression of GPR173 in excitatory neurons suppressed kainic acid (KA)-induced seizures in mice via the synthetic virus CaMKII-GPR173-mCherry.
From: Upregulation of CCK3R leading to the suppression of chronic seizures
A Experimental diagram showing the surgery timeline, intrahippocampal KA injection, epilepsy observation, AAV injection, and histology. After 3 weeks of intrahippocampal KA injection, 2 weeks of seizure observation were performed (Before). After intravenous viral (rAAV-CaMKII-GPR173-mCherry or rAAV-CaMKII-Sham-mCherry) delivery, the animals were continuously monitored for seizures for 8 weeks (After). B Heat maps of the number of seizures detected daily in the KA-injected mice (upper panel, N = 7), CaMKII-Sham-mCherry injected mice (middle panel, N = 8), and CaMKII-GPR173-mCherry injected mice (lower panel, N = 7). C Box chart for the average number of seizures per mouse within 2 weeks Before and After therapy (per week, total 8 weeks; KA group, N = 7, *P < 0.05; CaMKII-Sham-mCherry group, N = 8; CaMKII-GPR173-mCherry group, N = 7; one-way ANOVA with Tukey’s post hoc test, **P < 0.01; N.S.; one-way ANOVA with Tukey’s post hoc test, ###P < 0.001; N.S.). D Confirmation of mCherry reporter gene expression in the intravenously viral-injected chronic epileptic mice after the observation of seizures (left panel: CaMKII-GPR173-mCherry group; right panel: CaMKII-Sham-mCherry group; scale bars: 50 µm, respectively). E Percentage of mCherry and GPR173 co-labeling in the two groups (CaMKII-GPR173-mCherry group: N = 3 mice; CaMKII-Sham-mCherry group: N = 3 mice). F Confocal images of mCherry reporter gene expression with CaMKII in the cortex of CaMKII-GPR173-mCherry (left panels) or CaMKII-Sham-mCherry injected mice (right panels; scale bars: 50 µm). G Group data of CaMKII and mCherry co-labeling in the two groups. The transfection specificity means the proportion of CaMKII immunopositivity in the mCherry-expressing neurons, the transfection efficiency represents the proportion of CaMKII neurons transfected by the AAV (CaMKII-GPR173-mCherry group: N = 3 mice; CaMKII-Sham-mCherry group: N = 3 mice; one-way ANOVA with Tukey’s post hoc test, N.S.). Data are expressed as mean ± standard error of the mean. AAV adeno-associated virus, N.S. not significant, ANOVA analysis of variance.
