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. Author manuscript; available in PMC: 2021 Jan 27.
Published in final edited form as: Nat Metab. 2020 Jul 27;2(8):763–774. doi: 10.1038/s42255-020-0229-2

Figure 5. Arc GABA+ neuron activation driven by NachBac overrides leptin action.

Figure 5.

a, Diagram showing i.c.v. treatment of leptin or saline with 3-week duration minipumps in Vgat-Cre or Vgat-Cre::ob/ob mice that were injected with NachBac vectors to bilateral Arc. b-e, Body weight (b) and food intake (c) were measured once every 3 days with leptin or saline treatment in the indicated groups of mice and body composition was examined at the 18th day on leptin treatment for fat (d) and lean (e) mass, n=3 (NachBac+PBS), 3 (ob/ob+leptin) or 5 (NachBac+leptin), 2-way ANOVA in b (F(6, 35)=10.86, **p<0.0001, NachBac+PBS vs ob/ob+leptin; F(6, 42)=0.1529, p<0.9874, NachBac+PBS vs ob/ob+leptin; F(6, 49)=17.57, **p<0.0001, ob/ob+leptin vs NachBac+leptin) and in c (F(6, 35)=112.1, **p<0.0001, NachBac+PBS vs ob/ob+leptin; F(6,42)=6.368, p=0.081, NachBac+PBS vs ob/ob+leptin; F(6, 69)=290.2, **p<0.0001, ob/ob+leptin vs NachBac + leptin); and 2-sided unpaired Student’s t tests in d (t=10.40, df=4 **p=0.0005 in the ob/ob+leptin group (before vs after leptin)) and e (t=1.491, df=8, p=0.1778 in the group of NachBac+leptin (before vs after leptin)). f, Comparison in percentage of p-STAT3 expression in Arc GABA+ neurons in control and NachBac mice with saline or leptin infusion, n=3 each/group, 1-way ANOVA (F(3,8)=99.83, **p<0.0001, ob/ob+PBS vs ob/ob+leptin; p=0.9732, Control+leptin vs NachBac+leptin). g, Representative pictures for data presented in panel f showing GFP and p-STAT3 in Vgat-Cre::ob/ob mice with saline or leptin infusion (top 2 rows), and showing NachBac and p-STAT3 in NachBac injected Vgat-Cre mice with leptin infusion (bottom 2 rows). h-I, In a new cohort of Vgat-Cre male mice with NachBac viral delivery to the Arc, i.p. leptin (or saline) was administered, and the expression of p-STAT3 (h, t=11.71, df=4, **p=0.0003) and c-Fos (i, t=0.2800, df=4, p=0.754) was examined in the Arc and compared between leptin and contral saline treatments, n=3 each/group,with 2-sided unpaired Student’s t tests. j, Comparison in leptin effects on firing frequency of control and NachBac-expressing AgRP neurons using patch clamp recording on brain slices, n=11 (control) or 12 (NachBac), with 2-sided paired Student’s t tests, t=3.630, df=10, **p=0.0046 in controls, and t=0.8136, df=10, p=0.4348 in the NachBac group. 3V: the third ventricle. All data presented as mean +/− sem.