Extended Data Fig. 7: Functional validation of AAV-DIO-hGLP1R and AAV-DIO-mGLP1RS33W-HA and electrophysiological validation of AAV-DIO-hGLP1R in amygdala neurons.
From: A brain reward circuit inhibited by next-generation weight-loss drugs in mice
a, Schematic of Glp1r-Cre mice injected with a Cre-dependent AAV carrying full-length mouse Glp1r (mGlp1r) or AAV carrying full-length mouse GLP1R with the S33W mutation at position 33 (mGLP1RS33W) in the CeA. b,c, Normalized 4-hour (b) SD and (c) HFD consumption post-injection of vehicle or danuglipron in mGlp1r, full-length human GLP1R (hGLP1R) or mGlp1r-S33W (S33W) expressing mice mGLP1R n = 8, hGLP1R n = 9, mGLP1RS33W n = 4, two-way ANOVA with Bonferroni correction). d, Representative image of AAV-DIO-mGLP1RS33W-HA expression in the CeA (green). Scale bars, 200 µm. e–g, Localization of eYFP-labelled hGLP1R-expressing neurons in the amygdala of Glp1r-Cre mice co-injected with AAV-DIO-hGLP1R and AAV-DIO-eYFP (scale bars, 100 μm). h,i, Representative traces showing the effect of danuglipron perfusion (Dan; 30 μM) on the resting membrane potential of h, control mGlp1r-expressing neurons from Glp1r-Cre mice injected with AAV-DIO-mGlp1r and AAV-DIO-eYFP, and i, hGLP1R-expressing neurons. j, Average depolarization induced by danuglipron in mGlp1r- or hGLP1R-expressing neurons (mGlp1r: 13 cells from 7 mice; hGLP1R: 10 cells from 7 mice, unpaired t-test). k, Resting membrane potential of hGLP1R-expressing neurons before (baseline) and after 30 μM danuglipron perfusion (10 cells, 7 mice, paired t-test). l, Representative action potential traces from a hGLP1R-expressing neuron evoked by a slow current injection ramp (100 pA/s) before and after danuglipron perfusion. m, Time to threshold quantification of hGLP1R-expressing neurons before and after danuglipron perfusion (5 cells, 4 mice, paired t-test). Data are mean ± s.e.m. *P < 0.05; **P < 0.01; ***P < 0.001.
