Journal: bioRxiv
Article Title: The tongue-brain axis mediates a hidden amino acid appetite
doi: 10.64898/2026.04.17.719133
Figure Lengend Snippet: a, Schematics illustrating PVT injection of Saclofen, CGP55845 or PBS, and the timeline of diet feeding as well as the test. Mice were fed a (-) Leu diet for 3 days followed by medicine injection. b, Touch number percentage for the first 10 min of two-choice assays in a. c, Two-choice preferences for indicated time (10min, 30min, 1h, 2h) in a. d, Schematics illustrating the fiber photometry recordings of GABA signals in PVT neurons and the timeline of diet feeding as well as the test. Each mouse was subjected to the experiment twice. e, Heatmap of the fluorescence signals in the PVT neurons of mice that fed a Cont or (-) Leu diet for 3 days in response to a Cont diet. Each heatmap represents a single behavioral session. f, Averaged traces of fluorescence signals in e. g, The area under curve (AUC) of the fluorescence signals (0-600s) in f. h, Schematics illustrating virus-mediated shVGAT expression (red) in ARC AgRP neurons and the timeline of diet feeding as well as the test. Mice were fed a (-) Leu diet for 3 days. i, Touch number percentage for the first 10 min of two-choice assays in h. j, Two-choice preferences for indicated time (10min, 30min, 1h, 2h) in h. k, Schematics illustrating virus-mediated shVGAT expression (red) in ARC AgRP neurons, the fiber photometry recordings of GABA signals in PVT neurons, and the timeline of diet feeding as well as the test. Each mouse was subjected to the experiment twice. l, Heatmap of the fluorescence signals in the PVT neurons of mice that fed a Cont or (-) Leu diet for 3 days in response to a Cont diet. Each heatmap represents a single behavioral session. m, Averaged traces of fluorescence signals in l. n, The area under curve (AUC) of the fluorescence signals (0-600s) in m. o, Schematics illustrating virus-mediated shVGAT expression (red) in ARC AgRP neurons, the PVT injection of Baclofen or PBS, and the timeline of diet feeding as well as the test. Mice were fed a (-) Leu diet for 3 days. p, Touch number percentage for the first 10 min of two-choice assays in o. q, Two-choice preferences for indicated time (10min, 30min, 1h, 2h) in o. Studies for a-c were conducted using 10-12-week-old male WT mice with PVT injection of Saclofen, CGP55845 or PBS, fed a (-) Leu diet for 3 days; studies for d-g were conducted using 10-12-week-old male WT mice receiving AAVs expressing Syn-iGABASnFR fed a (-) Leu diet for 3 days; studies for h - j were conducted using 8-12-week-old female AgRP-Cre mice receiving AAVs expressing DIO-shNC or DIO-shVGAT in ARC fed a Cont or (-) Leu diet for 3 days; studies for k-n were conducted using 10-12-week-old male AgRP-Cre mice receiving AAVs expressing DIO-shNC or DIO-shVGAT in ARC, and Syn-iGABASnFR in PVT, fed a Cont or (-) Leu diet for 3 days; studies for o-q were conducted using 12-20-week-old female AgRP-Cre mice receiving AAVs expressing DIO-shNC or DIO-shVGAT in ARC, and PVT injection of Baclofen or PBS, fed a (-) Leu diet for 3 days. Data are expressed as the mean ± SEM (n = 6-20 per group, as indicated), with individual data points. Data were analyzed via two-tailed unpaired Student’s t-test (b, g, i, j, p), or one-way ANOVA followed by Dunnett’s multiple comparisons test (c), or two-way ANOVA followed by Tukey’s multiple comparisons test (n, q). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.
Article Snippet: The drugs including GABAbR antagonist saclofen (0.1 μg/μL, HY-100813 MCE) and CGP55845 hydrochloride (8 ng/μL, HY-103516, MCE, China), GABAbR agonist baclofen (0.04 μg/μL, HY-B0007 MCE), GABAaR antagonist bicuculline (2.5 μM, HY-N0219, MCE), MC3R/4R antagonist SHU 9119 (1 nM, HY-P0227, MCE), NPY1R antagonist BIBO3304 (0.25 mM, HY-107725, MCE), NPY2R antagonist BIIE-0246 (0.25 mM, HY-101986), NPY5R antagonist CGP71683 hydrochloride(15 nM, HY-107723, MCE) and leucine (1.1μg/μL in 10% DMSO).
Techniques: Injection, Fluorescence, Virus, Expressing, Two Tailed Test