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

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© 2023 The Authors

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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Fig. 2 — Increased and prolonged emotional reactivity to the optogenetic stimulation in vDG of GRov mice. (A) Experimental design. Mice expressing ChR2-eYFP were implanted in the vDG then tested for 18 min in the EPM with 6 min blue light off (Off1), 6 min blue light on (On), 6 min blue light off (Off2) epochs. (B) Expression of ChR2-eYFP in vDG. (C and D) Optogenetic activation of vDG induced a robust increase in the approach behavior in the EPM test in both GRov and WT mice. Repeated measures two-way ANOVA revealed a significant genotype × light interaction, F(2, 34) = 6.45, P < 0.01, for entries into the open arms in the EPM test. Repeated-measures one-way ANOVA revealed that blue light stimulation in vDG led to increased entries into the open arms in GRov [F(2, 20) = 36.92, P < 0.0001] and WT mice [F(2, 14) = 14.82, P < 0.001]. Repeated measures two-way ANOVA revealed there was no genotype × light interaction, F(2, 34) = 1.77, P = 0.19, for time spent in open arms in EPM test. Repeated-measures one-way ANOVA revealed that blue light stimulation in vDG led to increased time spent in the open arms in GRov [F(2, 20) = 76.5, P < 0.0001] and WT mice [F(2, 14) = 30.28, P < 0.0001]. Box plots show median, mean (+), lower and upper quartiles (boxes), and minima and maxima (whiskers). (E) Repeated measures two-way ANOVA revealed there was no genotype × light interaction, F(2, 34) = 2.61, P = 0.09, for total distance traveled in the EPM. Repeated-measures one-way ANOVA revealed that blue light stimulation in vDG led to increased distance traveled in EPM in GRov [F(2, 20) = 31.26, P < 0.0001] and WT mice [F(2, 14) = 14.82, P < 0.001]. GRov, n = 11 mice; WT, n = 8 mice. **P < 0.01, ***P < 0.001, ****P < 0.0001 versus respective pre-stimulation group. (F) Following optogenetic stimulation in vDG, GRov mice exhibited a greater response range in entering into open arms than WT. Unpaired two tailed t-test: t(17) = 7.17, ****P < 0.0001. (G) Moreover, GRov mice continued to display a greater response range in entering open arms after the termination of light stimulation than WT. Unpaired two tailed t-test: t(17) = 4.02, ***P < 0.001. (H) Following optogenetic stimulation in vDG, GRov mice exhibited a greater response range in time spent in open arms than WT. Unpaired two tailed t-test: t(17) = 6.77, ****P < 0.0001. (I) GRov mice continued to display a greater response range in time spent in open arms after the termination of light stimulation than WT mice. Unpaired two tailed t-test: t(17) = 2.88, *P < 0.05. (J) There is no genotype difference in response range in distance traveled in EPM following the blue laser stimulation. Unpaired two tailed t-test: t(17) = 0.54, P = 0.6. (K) GRov mice displayed a greater response range in distance traveled in EPM after the termination of light stimulation. Unpaired two tailed t-test: t(17) = 2.46, *P < 0.05. GRov, n = 11 mice; WT, n = 8 mice.