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. Author manuscript; available in PMC: 2020 May 8.
Published in final edited form as: Neuron. 2019 Mar 14;102(3):653–667.e6. doi: 10.1016/j.neuron.2019.02.028

Figure 1. PVHPDYN neurons are a MC4R-negative satiety population that accounts for the missing 50% of PVHSIM1 neuron-mediated satiety.

Figure 1.

(A) Schematic of Mc4r-T2A-Cre::Ai9-Rosa26-LSL-TdTom::Pdyn-EGFP mouse. (B) Representative histological sections of the PVH from rostral to caudal (MC4R::TdTOM=red, PDYN-EGFP=green, double labeled=yellow) (C) Quantification MC4R::TdTOM+, PDYN-EGFP+ vs. double labeled neurons summed across the rostra-caudal extent of the PVH (n=4). (D) Schematic of AAV-DIO-hM4Di injection in the PVH of Sim1-IRES-Cre, Mc4r-T2A-Cre, Pdyn-IRES-Cre, or compound Pdyn-IRES-Cre::Mc4r-T2A-Cre mice. (E) Representative images of hM4Di-mCherry expression in PVHSIM1, PVHMC4R, PVHPDYN, and PVHPDYN::MC4R neurons in the PVH. (F) Light-cycle cumulative food intake in sated mice following saline vs. CNO/hM4Di inhibition of PVHSIM1, PVHMC4R, PVHPDYN and PVHPDYN::MC4R neurons. PVHSIM1 n=6: Repeated Measures (RM) 2 way ANOVA Time F(3,15)=87.43 ****P<0.0001, Treatment F(1,5)=243.8 ****P<0.00001, Interaction F(3,15)=57.22. Sidak’s Multiple Comparisons: Saline vs. CNO ****P<0.00001 at 1,2,3 hours. PVHMC4R n=7: RM 2 way ANOVA Time F(3,18)=23.47 ****P<0.0001. Treatment F(1,6)=109.4 ****P<0.0001, Interaction F(3,18)=23.01, ****P<0.0001, Sidak’s Multiple Comparisons: Saline vs. CNO **P<0.01 at hour 1, ****P<0.0001 at hour 2,3. PVHPDYN n=5: RM 2 way ANOVA Time F (3,12)=26.03 ****P<0.0001, Treatment F(1,4)=50.07 ***P<0.001, Interaction F(3,12)=34.47 ****P<0.0001. Sidak’s Multiple Comparisons: Saline vs. CNO ****P<0.0001 at hours 1,2,3. PVHPDYN::MC4R n=5: RM 2 way ANOVA Time F(3,12)=135. 4 ****P<0.0001, Treatment F(1,4)=245.9 *****P<0.0001, Interaction F(3,12)=88.51 ****P<0.0001, Sidak’s Multiple Comparisons: Saline vs. CNO ****P<0.0001 at hours 1,2,3. (G) Same data at the 3 hour time point quantified as change in food intake (CNO-Saline). ANOVA F(3, 18) = 25.87 ****P<0.0001, Tukey’s Multiple Comparisons between groups: PVHSIM1 vs. PVHMC4R ****P<0.0001, PVHSIM1 vs. PVHPDYN ****P>0.0001, PVHMC4R vs. PVHPDYN:MC4R ***P<0.001, PVHPDYN vs. **PVHPDYN::MC4R P<0.01, PVHPDYN::MC4R vs. PVHSIM1 N.S., PVHMC4R vs. PVHPDYN NS. Data are represented as mean ± SEM.