Fig. 8: Chemogenetic activation and tetanus toxin-mediated synaptic silencing of CCK-expressing neurons in DMH. | Nature Communications

Fig. 8: Chemogenetic activation and tetanus toxin-mediated synaptic silencing of CCK-expressing neurons in DMH.

From: A sex-specific thermogenic neurocircuit induced by predator smell recruiting cholecystokinin neurons in the dorsomedial hypothalamus

Fig. 8

a Experimental design for chemogenetic manipulation of CCK-expressing neurons in the DMH. b Bilateral stereotaxic targeting of DMH neurons using AAV-DIO-hM3D-mCherry shows robust mCherry signal in the d/cDMH overlapping with cFos after CNO injection, Two independent experiments, N = 15. c Oxygen consumption in CCK-IRES-Cre+ mice after CNO injection, males, N = 4 per group, females, N = 3/mCherry and N = 4/hM3D. d Average VO2 in CCK-IRES-Cre+ measured for 3 h post-CNO injection, males, N = 4 per group, females, N = 3/mCherry and N = 4/hM3D, Unpaired t-test,Two-tailed. e Total activity in CCK-IRES-Cre+ mice after CNO injection, males, N = 4 per group, females, N = 3/mCherry and N = 4/hM3D. f Food intake measured after overnight fasting and upon CNO injection, males, N = 4 per group, females, N = 3/mCherry and N = 4/hM3D, Two-way Anova with Sidak’s post hoc comparison. g CORT levels post-CNO injection, N = 4 per group. h Experimental design for tetanus toxin-mediated synaptic silencing of CCK-expressing neurons in the DMH. i eGFP positive cells in cDMH after stereotaxic AAV injection, Two independent experiments, N = 16. j VO2 measured upon TMT or control swab exposure, N = 8 per group, Three-way Anova. k VO2 before the stimulus (t = 0 min) and after exposure to the stimulus (t = 22.5 min), N = 8 per group, Two-way Anova with Tukey’s post hoc comparison. l Activity measured upon TMT or control swab exposure, N = 8 per group, Three-way Anova. m Activity before the stimulus (t = 0 min) and after exposure to the stimulus (t = 22.5 min), N = 8 per group, Two-way Anova with Tukey’s post hoc comparison. n Food intake measured after overnight fasting and upon TMT or control swab exposure, eGFP, N = 7/No odor and N = 6/TMT, TeNT, N = 8/No odor and N = 7/TMT, Three-way Anova. o Cumulative food intake after 6 h of refeeding under TMT or control swab exposure, eGFP, N = 7/No odor and N = 6/TMT, TeNT, N = 8/No odor and N = 7/TMT, Two-way Anova with Tukey’s post hoc comparison. All bar graphs are presented as mean values ± SEM. Table S7 contains the detailed results of the statistical analysis. *p < 0.05, **p < 0.01, ****p  < 0.0001. Source data are provided as a Source Data file. Clozapine-N-Oxide (CNO), 2,4,5-trimethylthiazoline (TMT), corticosterone (CORT), compact dorsomedial hypothalamus (cDMH), Adeno-associated viruses (AAV).

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