Fig. 7
From: Neurocircuitry underlying the antidepressant effect of retrograde facial botulinum toxin in mice
Excitation of wFMNs-projecting vlPAG excitatory neurons attenuates the antidepressant-like behavior of facial BoNT/A. a Schematic of experimental design. b Scheme for specific infection of wFMNs-projecting vlPAG excitatory neurons with hM3Dq or mCherry by stereotaxic infusion of rAAV-retro-CaMKIIα-EGFP-Cre into the unilateral lFN at the site of wFMNs and AAV-DIO-hM3Dq-mCherry or AAV-DIO-mCherry into the ipsilateral vlPAG. c Representative images of lFN (left) and vlPAG (right) 2 weeks after virus injection. Scale bar, 500 μm. d Representative image of the vlPAG indicated that i.p. injection of CNO evoked c-Fos expressions in neurons expressing hM3Dq. Scale bar, 100 μm. e Behavioral despair in the FST (left and middle) and anhedonia in the SPT (right) among experimental groups. All mice injected by a rAAV-retro-CaMKIIα-EGFP-Cre vector into unilateral lFN and exposure to CRS. Saline + hM3Dq + Saline, CRS mice that received WIM pre-injection of saline, vlPAG injection of AAV-DIO-hM3Dq-mCherry, and i.p. injection of saline. Saline + hM3Dq + CNO, CRS mice that received WIM pre-injection of saline, vlPAG injection of AAV-DIO-hM3Dq-mCherry, and i.p. injection of CNO. BoNT/A + mCherry + CNO, CRS mice that received WIM pre-injection of BoNT/A, vlPAG injection of AAV-DIO-mCherry, and i.p. injection of CNO. BoNT/A + hM3Dq + Saline, CRS mice that received WIM pre-injection of BoNT/A, vlPAG injection of AAV-DIO-hM3Dq-mCherry, and i.p. injection of saline. BoNT/A + hM3Dq + CNO, CRS mice that received WIM pre-injection of BoNT/A, vlPAG injection of AAV-DIO-hM3Dq-mCherry, and i.p. injection of CNO. n = 8 animals from the group of Saline + hM3Dq + Saline, Saline + hM3Dq + CNO and BoNT/A + mCherry + CNO, n = 9 animals from the group of BoNT/A + hM3Dq + Saline and BoNT/A + hM3Dq + CNO. One-way ANOVA followed by Bonferroni’s multiple comparisons test, F (4, 37) = 7.705, P = 0.0001 for latency to immobility in the FST; F (4, 37) = 6.778, P = 0.0003 for immobility duration in the FST; F (4, 37) = 8.012, P < 0.0001 for sucrose preference in the SPT. The P-value of Bonferroni’s multiple comparisons: Saline + hM3Dq + Saline vs. BoNT/A + mCherry + CNO: P = 0.0199, Saline + hM3Dq + Saline vs. BoNT/A + hM3Dq + Saline: P = 0.0244, Saline + hM3Dq + CNO vs. BoNT/A + mCherry + CNO: P = 0.0063, Saline + hM3Dq + CNO vs. BoNT/A + hM3Dq + Saline: P = 0.0076, BoNT/A + mCherry + CNO vs. BoNT/A + hM3Dq + CNO: P = 0.0063, BoNT/A + hM3Dq + Saline vs. BoNT/A + hM3Dq + CNO: P = 0.0075 of latency of immobility; Saline + hM3Dq + Saline vs. BoNT/A + mCherry + CNO: P = 0.0084, Saline + hM3Dq + Saline vs. BoNT/A + hM3Dq + Saline: P = 0.0049, Saline + hM3Dq + CNO vs. BoNT/A + mCherry + CNO: P = 0.0421, Saline + hM3Dq + CNO vs. BoNT/A + hM3Dq + Saline: P = 0.0265, BoNT/A + mCherry + CNO vs. BoNT/A + hM3Dq + CNO: P = 0.0458, BoNT/A + hM3Dq + Saline vs. BoNT/A + hM3Dq + CNO: P = 0.0284 of total time of immobility; Saline + hM3Dq + Saline vs. BoNT/A + mCherry + CNO: P = 0.0028, Saline + hM3Dq + Saline vs. BoNT/A + hM3Dq + Saline: P = 0.0034, Saline + hM3Dq + CNO vs. BoNT/A + mCherry + CNO: P = 0.0258, Saline + hM3Dq + CNO vs. BoNT/A + hM3Dq + Saline: P = 0.0323, BoNT/A + mCherry + CNO vs. BoNT/A + hM3Dq + CNO: P = 0.0068, BoNT/A + hM3Dq + Saline vs. BoNT/A + hM3Dq + CNO: P = 0.0083 of sucrose preference. *P < 0.05, **P < 0.01