Fig. 4: DREADD-dependent activation of POMCLepr+ or POMCGlp1r+ neurons differentially reduces food intake. | Nature Neuroscience

Fig. 4: DREADD-dependent activation of POMCLepr+ or POMCGlp1r+ neurons differentially reduces food intake.

From: Functionally distinct POMC-expressing neuron subpopulations in hypothalamus revealed by intersectional targeting

Fig. 4

a, Illustrations of experimental mice and schematic diagram showing Dre- and Cre-dependent targeted expression of activatory hM3Dq in either POMCLepr+ or POMCGlp1r+ neurons. Excision of loxP-flanked and rox-flanked stop cassettes through recombination of both Dre and Cre drivers leads to hM3Dq expression in the targeted subpopulation. b, Representative microscopic images of RNA ISH against Pomc, Lepr, ZsGreen (in lieu of hM3Dq) and Fos in POMCDre LeprCre ROSA26lSlrSrhM3Dq males injected with saline or CNO. Images on the left show ISH in the ARC with nuclear counterstain (blue, DAPI). Magnifications of the boxes (right) are shown with the indicated stainings. Pomc-positive neurons are outlined in white. Scale bars, 50 μm (merged image) and 25 μm (magnified images). c–e, Percentage of ZsGreen-Pomc-positive cells expressing Lepr or Glp1r (c), percentage of Lepr/Glp1r-Pomc-positive cells expressing ZsGreen (d) and percentage of ZsGreen-Pomc-positive cells expressing Fos (e) in POMCDre LeprCre ROSA26lSlrSrhM3Dq or POMCDre Glp1rCre ROSA26lSlrSrhM3Dq male mice (22–26 weeks old) injected with saline or CNO. CNO, 3 mg kg−1. c: POMCLepr+: saline: 91.84% ± 1.03%, CNO: 94.87% ± 1.62%, saline versus CNO, t = 1.580, P = 0.342863: POMCGlp1r+: saline: 93.52% ± 2.04%, CNO: 96.16% ± 2.31% saline versus CNO, t = 0.8592, P = 0.438695; d: POMCLepr+: saline: 45.46% ± 6.92%, CNO: 49.19% ± 1.92%, saline versus CNO, t = 0.5191, P = 0.631109; POMCGlp1r+: saline: 36.88% ± 5.28%, CNO: 43.24% ± 1.42%, saline versus CNO, t = 1.165, P = 0.522274. e: POMCLepr+: saline: 6.87% ± 2.64%, CNO: 94.03% ± 1.66%, saline versus CNO, t = 28.0, df = 4, PuT = 0.000019; POMCGlp1r+: saline: 8.23% ± 1.13%, CNO: 87.05% ± 6.05%, saline versus CNO, t = 12.82, df = 4, PuT = 0.000214, unpaired Student’s t-test, Holm–Sidak correction; n = 3 mice. f,g, Food intake over a time course of 24 h in POMCDre LeprCre ROSA26lSlrSrhM3Dq (f) and POMCDre Glp1rCre ROSA26lSlrSrhM3Dq male mice (g) starting with the night cycle. Mice were injected with saline at 18:00 and 23:00, followed by a 1-d gap and subsequent CNO injections at 18:00 and 23:00 on the next day. Left: cumulative food intake in mice injected with saline versus CNO; right, total food intake in grams during night and day. f: n = 8; left: saline versus CNO two-way ANOVA, F(1,7) = 4.815, P = 0.0643; right: salineNight: 3.03 ± 0.39, CNONight: 2.79 ± 0.20, saline versus CNO two-way ANOVA followed by Sidak’s test, P = 0.2729, salineDay: 1.05 ± 0.15, CNODay: 1.10 ± 0.07, saline versus CNO two-way ANOVA followed by Sidak’s test, P = 0.9867. g: n = 7, left: saline versus CNO two-way ANOVA, F(1.000, 6.000) = 16.51, P = 0.0066, right: salineNight: 3.42 ± 0.38, CNONight: 2.32 ± 0.12, salineDay: 1.19 ± 0.15, CNODay: 0.98 ± 0.16; saline versus CNO two-way ANOVA followed by Sidak’s test, P = 0.0312; saline versus CNO two-way ANOVA followed by Sidak’s test, P = 0.7992. Data are represented as the mean ± s.e.m. Statistical analyses in c–e were performed by unpaired two-tailed Student’s t-test with Holm–Sidak correction for multiple comparisons. For cumulative food intake (f and g left), two-way ANOVA was used; for total food intake (f and g right), two-way ANOVA followed by Sidak’s post hoc test was used. Indices PuT: unpaired t-test. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ***P ≤ 0.0001.

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