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. Author manuscript; available in PMC: 2018 Sep 27.
Published in final edited form as: Neuron. 2017 Sep 27;96(1):190–206.e7. doi: 10.1016/j.neuron.2017.09.014

Figure 6. NTSHSD2 neuron stimulation synergizes with angiotensin II signaling to drive sodium appetite.

Figure 6

See also Figure S6. Data are presented as mean ± SEM.

A) Schematic of AAV-DIO-hM3Dq-mCherry injections.

B) Representative recording of an hM3Dq-expressing NTSHSD2 neuron pre- and post-CNO (5 µM) application.

C) NTS hM3Dq-mCherry expression in Hsd11b2-Cre mice (left), Fos immunoreactivity (Fos-IR; middle), and hM3Dq-mCherry + Fos-IR (right) in vehicle (top) or CNO (bottom) treated mice.

D) 3% NaCl and H2O licking behavior over 60 minutes after CNO/hM3Dq stimulation of NTSHSD2 neurons in euhydrated mice (n = 18 mice).

E) Time course of 3% NaCl licking (left) or H2O licking (right) following CNO/hM3Dq stimulation of NTSHSD2 neurons in H2O-restricted mice.

F) Summary of 60 minutes of licking behavior following CNO/hM3Dq stimulation of NTSHSD2 neurons in H2O-restricted mice (n = 11 mice). Two-way repeated measures ANOVA followed by Tukey’s multiple comparisons test, *P < 0.05, **P < 0.01, ***P < 0.001.

G) 3% KCl and H2O licking behavior over 60 minutes following chemogenetic activation of NTSHSD2 neurons in H2O-restricted mice (n = 8 mice).

H) Plasma ATII levels (fold change) under control, furosemide + LSD (Furo), and H2O-restricted (H2O-R) conditions (n = 6 mice/group). One-way ANOVA with posthoc analysis by Dunnett's multiple comparisons test, *P < 0.05, **P < 0.01.

I) 3% NaCl (left) and H2O (right) licking behavior over 60 minutes following CNO/hM3Dq stimulation of NTSHSD2 neurons in H2O-restricted animals ± the AT1aR antagonist losartan (20 mg/kg; n = 14 mice). Two-way repeated measures ANOVA followed by Tukey’s multiple comparisons test, **P < 0.01, ***P < 0.001.

J) 3% NaCl and H2O licking behavior over 60 minutes after CNO/hM3Dq stimulation of NTSHSD2 neurons and injection of 1 M NaCl (n = 8 mice).

K) 3% NaCl and H2O licking behavior over 60 minutes after CNO/hM3Dq stimulation of NTSHSD2 neurons and injection of 1 M sucrose (n = 7 mice).

L) 3% NaCl (left) and H2O (right) licking behavior over 60 minutes following CNO/hM3Dq stimulation of NTSHSD2 neurons under euhydrated conditions ± ATII injections (0.25 mg/kg; n = 8 mice). Two-way repeated measures ANOVA followed by Tukey’s multiple comparisons test, ****P < 0.0001.