Extended Data Fig. 1: Assessment of chemogenetic modulation of AgRP and POMC neurons.
From: Rapid modulation of gut microbiota composition by hypothalamic circuits in mice
a–d. Schematic of the approach to express the excitatory (hM3Dq) or inhibitory DREADDs (hM4Di) in AgRP and POMC neurons (n = 3 mice per group). The representative images show mCherry reporter expression in ARC coronal sections. a, b. Assessment of chemogenetic activation (A) or inhibition (B) of AgRP neurons in AgRP+/+ and AgRPCre/+ mice expressing hM3Dq or hM4Di two hours after CNO injection. Representative images of FOS (green), mCherry (red), and DAPI nuclei counterstaining (blue) are shown. Scale bar: 50 μm. c, d. Assessment of chemogenetic activation (C) or inhibition (D) of POMC neurons in POMC+/+ and POMCCre/+ mice expressing hM3Dq or hM4Di two hours after CNO injection. Representative images of FOS (magenta) and POMC (green) immunostaining, mCherry (red) and DAPI nuclei counterstaining (blue) are shown. Scale bar: 50 μm. e, f. Quantification of FOS-positive cells in AgRP+/+ and AgRPCre/+ mice expressing hM3Dq (n = 3 mice per group) (E; *p = 0.0205 hM3Dq, 2 h; **p = 0.0015 hM3Dq, 4 h) or hM4Di (F; *p = 0.0162 hM4Di, 2 h; *p = 0.0274 hM4Di, 4 h) two or four hours after CNO injection. g, h. Quantification of FOS-positive POMC neurons in POMC+/+ and POMCCre/+ mice expressing hM3Dq (n = 3 mice per group) (G; ****p < 0.0001 hM3Dq, 2 h; ****p < 0.0001 hM3Dq, 4 h) or hM4Di (H; **p = 0.0022 hM4Di, 2 h; **p = 0.037 hM4Di, 4 h) two or four hours after CNO injection. Statistical significance was determined with a two-tailed t test. hM3Dq: AAV8-hSYN-DIO-hM3D(Gq)mCherry; hM4Di: AAV8-hSYN-DIO-hM4D(Gi)mCherry; CNO: Clozapine-N-Oxide. Data in panels E-H are represented as mean ± SEM.
