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. Author manuscript; available in PMC: 2021 Feb 5.
Published in final edited form as: Neuron. 2019 Nov 25;105(3):475–490.e6. doi: 10.1016/j.neuron.2019.10.035

Figure 7. Chemogenetic inhibition of vHPC pyramidal neurons attenuated anxiety-like behaviors in NEX-Cul3f/+ mice.

Figure 7.

(A) Diagram of Cre-DIO-dependent expression (left) and validation of hM4Di-mCherry in vHPC in NEX-Cul3f/+ mice. Scale bar, 1 mm.

(B) Representative spike traces of vHPC CA1 pyramidal neurons. hM4Di or mCherry expressing brain slices were incubated with 10 µM CNO and neurons were stimulated by injected depolarizing currents at 50 pA, 150 pA and 300 pA.

(C) CNO reduced RMP of hM4Di-expressing neurons. n = 11 neurons, 3 mice for all groups; for NEX-Cul3f/+-mCherry+Veh (−65.5 ± 1.2 mV) vs NEX-Cul3f/+-mCherry+CNO (−65.1 ± 1.4 mV); p > 0.9999; for NEX-Cul3f/+-mCherry+CNO vs NEX-Cul3f/+-hM4Di+CNO (−70.9 ± 1.9 mV); p = 0.0033; Kruskal-Wallis ANOVA followed by Dunn’s post hoc test.

(D) CNO (2 mg/kg, i.p.) increased time spent in center of hM4Di-injected NEX-Cul3f/+ mice during open field tests. n = 12 mice for all groups; for NEX-mCherry + Veh (231 ± 25.7 s) vs NEX-Cul3f/+-mCherry + Veh (122 ± 24.4 s), p = 0.0078; NEX-Cul3f/+-mCherry + CNO (118 ± 18.3 s) vs NEX-Cul3f/+-hM4Di + CNO (224 ± 19.7 s), p = 0.0329; Kruskal-Wallis ANOVA followed by Dunn’s post hoc test.

(E) No effect by CNO on overall distance traveled of indicated groups. n = 12 mice for all groups; p > 0.9999 for all comparisons; Kruskal-Wallis ANOVA followed by Dunn’s post hoc test.

(F) CNO increased time spent in open arms of hM4Di-injected NEX-Cul3f/+ mice in EPM tests. n = 12 mice for all groups; for NEX-mCherry + Veh (65.2 ± 8.4 s) vs NEX-Cul3f/+-mCherry + Veh (38.1 ± 6.9 s), p = 0.0268; NEX-Cul3f/+-mCherry + CNO (35.4 ± 3.7 s) vs NEX-Cul3f/+-hM4Di + CNO (61.6 ± 7.4 s), p = 0.0419; Kruskal-Wallis ANOVA followed by Dunn’s post hoc test.

(G) CNO increased number of entries of hM4Di-injected NEX-Cul3f/+ mice in EPM tests. n = 12 mice for all groups; for NEX-mCherry + Veh (11.4 ± 0.8) vs NEX-Cul3f/+-mCherry + Veh (6.2 ± 0.9), p = 0.0092; NEX-Cul3f/+-mCherry + CNO (5.1 ± 0.4) vs NEX-Cul3f/+-hM4Di + CNO (11.6 ± 0.8), p = 0.0002; Kruskal-Wallis ANOVA followed by Dunn’s post hoc test.

Data were shown as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001; ns, no significant difference.