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. 2021 Feb 2;10:e63671. doi: 10.7554/eLife.63671

Figure 1. Silencing DMHLepR neurons elicits transient hyperphagia and increased adiposity in adult male mice.

(A) Experimental schematic for chronic inhibition of DMHLepR by microinjection on day 0 of an AAV1 containing a Cre-dependent GFP-fused TeTx delivered bilaterally to the DMH of LepR-Cre+ male mice (TeTx; n=7) and Cre-negative littermate controls (control; n=5). (B) Stereological fluorescent images from a representative animal showing the rostral-caudal extent of GFP:TeTx expression. (C) Left: colorized, higher magnification view of the boxed orange region from (B). Middle: higher magnification view of the boxed orange region showing neuronal cell bodies targeted within the DMH. Right: higher magnification view of the boxed red region showing GFP:TeTx+ terminals of targeted DMHLepR neurons within the arcuate nucleus (ARC). (D) Mean daily food intake following viral microinjection. Two-way ANOVA: F(1,10)=4.658; p=0.0563 (main effect of TeTx); F(14,140)=4.886; p<0.0001 (time x TeTx interaction). (E) Mean daily food intake from week 1 relative to week 4. Two-way ANOVA: F(1,10)=5.575; p=0.0399 (main effect of TeTx); F(1,10)=39; p<0.001 (time x TeTx interaction). (F) Body weight expressed as %day 0 value. Two-way ANOVA: F(1,10)=20.18; p=0.0012 (main effect of TeTx). F(19,190)=14.67; p<0.0001 (time x TeTx interaction). (G) Fat, lean, and total mass 26 days after viral microinjection. Multiple t-tests; tfat=4.847; p=0.0014; ttotal=2.884; p=0.016. (H) Plasma leptin 21 days after viral microinjection. Unpaired t-test, t=5.17, p=0.0017. Data are mean ± SEM. For repeated measures, post hoc Sidak’s test for each time point is indicated on the graph. *p<0.05,**p<0.01, ***p<0.001, ****p<0.0001.

Figure 1—source data 1. Longitudinal measures following DMH-LepR TeTx.

Figure 1.

Figure 1—figure supplement 1. Representative viral expression is evident in both the ventral and dorsal compartments of the dorsomedial hypothalamic nucleus (DMH) following microinjection of GFP:TeTx to the DMH of LepR-Cre+ male mice.

Figure 1—figure supplement 1.

Animal #6 represents a surgical miss and was excluded from all analyses.
Figure 1—figure supplement 2. Silencing DMHLepR neurons in female mice recapitulates body weight and fat mass increase observed in males, but not acute hyperphagia.

Figure 1—figure supplement 2.

(A) Average daily food intake following chronic inhibition of DMHLepR by bilateral microinjection on day 0 of a Cre-dependent GFP:TeTx delivered to LepR-Cre+ female mice (TeTx; n=9) and Cre-negative littermate controls (control; n=5). Two-way ANOVA: F(1,12)=0.3474; p=0.5665 (main effect of TeTx); F(13,156)=1.563, p=0.1014 (time x TeTx interaction). (B) Cumulative food intake for the first 11 days in A (inset). Two-way ANOVA: F(1,12)=1.503; p=0.2437 (main effect of TeTx); F(10,120)=1.427, p=0.1766 (time x TeTx interaction). (C) Daily body weight expressed as %day 0 value. Two-way ANOVA: F(1,12)=7.11; p=0.0205 (main effect of TeTx); F(14,168)=15.83, p<0.0001 (time x TeTx interaction). (D) Fat, lean, and total mass 14 days after viral microinjection. Multiple t-tests; tfat=3.268; p=0.0024; ttotal=4.705; p<0.0001. Data are mean ± SEM. *p<0.05,**p<0.01, ***p<0.001, ****p<0.0001.