Fig. 3: Optogenetic stimulation of the ARCPOMC → DMVACh → liver projection causes hepatic gluconeogenesis.
a Schematic diagram of the experimental configuration. AAV8-WGA-Cre viruses were injected into the liver and 2−3 weeks post viral injections, AAV5-nPE-ChR2-YFP viruses were injected into the ARC of C57BL/6J mice. The ARCPOMC → DMVACh → liver projection was optogenetically stimulated at 20 Hz for 1 h. b, c Pooled data showing that optogenetic stimulation of the ARCPOMC → DMVACh → liver projection increased blood glucose levels in male mice (control (laser off), n = 12 mice (open circle), stimulation (laser on), n = 15 mice (filled circle)), two-way RM ANOVA followed by Sidak multiple comparisons test, interaction F (5, 125) = 18.2, p < 0.001, time F (5, 125) = 21.4, p < 0.001, between the groups F (1, 25) = 24.1, p < 0.001; 30 min, ***p < 0.001; 45 min, ***p < 0.001; 60 min, ***p < 0.001 (b, left panel). Right panel: pooled data showing increased blood glucose levels at the end of optogenetic stimulation (two-tailed t test, ***p < 0.001, vs. 0 min, n = 15 mice). c Pooled data showing changes in blood glucose levels before and after 1-h optogenetic stimulation (two-tailed t test, ***p < 0.001, vs. 0 min, n = 15 mice). Mice were not fasted. All data are shown as mean ± SEM. Source data are provided as a Source Data file. d, e Pooled data showing effect of optogenetic stimulation of the ARCPOMC → DMVACh → liver projection on blood glucose in female mice (control (laser off), n = 10 mice (open circle), stimulation (laser on), n = 8 mice (filled circle)), two-way RM ANOVA followed by Sidak multiple comparisons test, interaction F (5, 80) = 18.5, p < 0.001, time F (5, 80) = 18.9, p < 0.001, between the groups F (1, 16) = 38.6, p < 0.001; 15 min, **p = 0.005; 30 min, ***p < 0.001; 45 min, ***p < 0.001; 60 min, ***p < 0.001 (d, left panel). Right panel: summary plot showing increased blood glucose levels before and after optogenetic stimulation (two-tailed t test, **p = 0.001, vs. 0 min, n = 8 mice). e Pooled data showing changes in blood glucose levels before and after 1-h optogenetic stimulation (two-tailed t test, **p = 0.001 vs. 0 min, n = 8 mice). Mice were not fasted. All data are shown as mean ± SEM. Source data are provided as a Source Data file. f Pooled data showing effect of SHU9119 on optogenetic stimulation-induced increase in blood glucose (saline, n = 5 mice, SHU9119, n = 5 mice, two-way RM ANOVA followed by Sidak multiple comparisons test, interaction F (5, 40) = 8.6, p < 0.001, time F (5, 40) = 4.2, p < 0.01, between the two groups, F (1, 8) = 7.5, p = 0.03; 45 min, *p = 0.01, 60 min, ***p < 0.001, left panel). Right panel: summary plot showing changes in blood glucose at the end of optogenetic stimulation with saline or SHU9119 (two-tailed t test, ***p < 0.001, vs. saline, n = 5 mice). Mice were not fasted. All data are shown as mean ± SEM. Source data are provided as a Source Data file. g Pooled data showing increased G6pase and Pepck mRNA expression in mice with optogenetic stimulation (two-tailed t test, ***p < 0.001, vs. control, control, n = 18 mice; stimulation, n = 8 mice). All data are shown as mean ± SEM. Source data are provided as a Source Data file. h–j Summary plots showing plasma glucagon (h), insulin (i), and corticosterone (j) levels following stimulation of the ARCPOMC → DMVACh → liver projection (two-tailed t test, glucagon, control, n = 15 mice, stimulation, n = 8 mice, p = 0.98; insulin, control, n = 16 mice, stimulation, n = 12 mice, p = 0.12; corticosterone, control, n = 12 mice, stimulation, n = 5 mice, p = 0.24). All data are shown as mean ± SEM. Source data are provided as a Source Data file.
