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. 2023 Oct 16;27:100581. doi: 10.1016/j.ynstr.2023.100581

Fig. 5.

Fig. 5

Glutamatergic/GABAergic activation imbalance in vCA1 of GRov mice following optogenetic stimulation in vDG. (A) Representative confocal photomicrographs show the increased cFos activity in the ventral hippocampus and the amygdala subregions following blue laser stimulation in vDG of WT and GRov mice compared to no stimulation groups. HCR FISH labeling is represented as cFos (red), Vglut1 (blue), Gad2 (green), ChR2 (cyan), cFos + Vglut1 (magenta), cFos + Gad2 (yellow). Scale bars, 500 μm. (B) Magnified views of vCA1 show cFos co-labeling with Vglut1+ and Gad2+. Majority of the cFos+Vglut1 (magenta) labeling can be seen in the pyramidal layer (Py) whereas cFos + Gad2 (yellow) neurons were mostly localized in the stratum oriens (Or), stratum radiatum (Rad) and stratum lacunosum-moleculare (Lm) layers of vCA1. cc- Corpus Callosum. In the representative merged images cFos + Vglut1 and cFos + Gad2 colocalizations are indicated by magenta and yellow arrows, whereas individual labeling of cFos, Vglut1 and Gad2 are indicated by red, blue and green arrows, respectively. Scale bars, 50 μm. (C) Histogram shows the percentage of cFos+Vglut1+ to total Vglut1+ neuronal number density in the vDG, vCA1 and vCA3 following the optogenetic stimulation of vDG in both GRov and WT groups compared to the no stimulation groups. Two-way ANOVA analyses were performed on listed regions, followed by Šídák's multiple comparisons test. Two-way ANOVA revealed a significant genotype × light interaction in vCA1 [F(1, 11) = 10.14, P < 0.01]. **P < 0.01, ***P < 0.001, ****P < 0.0001 versus respective no stimulation group; ##P < 0.01 versus the stimulated WT group. There was no genotype difference in cFos-based glutamatergic activity in vDG, vCA1, and vCA3 between GRov and WT mice under the no stimulation condition. Significantly higher cFos-based glutamatergic activity was observed in the vCA1 of stimulated GRov mice compared to stimulated WT animals. (D) Histogram shows the percentage of cFos+Gad2+ to total Gad2+ neuronal number density in the vDG, vCA1, and vCA3 following the optogenetic stimulation of vDG in both WT and GRov mice, compared to the no stimulation groups. Two-way ANOVA revealed a significant genotype × light interaction in vCA1 [F(1, 11) = 10.12, P < 0.01]. ****P < 0.0001 versus respective no stimulation group; ##P < 0.01 versus the stimulated WT group. There was no genotype difference in cFos-based GABAergic activity in vDG, vCA1, and vCA3 between GRov and WT mice under the no stimulation condition. However, in the vCA1 region significantly lower cFos-based GABAergic activity was observed in the stimulated GRov group compared to stimulated WT group. Mean ± SEM. WT (light off), n = 3 mice; GRov (light off), n = 3 mice; WT (light on), n = 4 mice; GRov (light on), n = 5 mice.