Supplementary Figure 6: CRACM mapping and validation of hM3Dq/ChR2 expression in ARCOxtr neurons.
From: A rapidly acting glutamatergic ARC→PVH satiety circuit postsynaptically regulated by α-MSH
a, Top, schematic shows connections being tested. Bottom, representative traces showing assessment of light-evoked IPSCs recorded from an uPVH neuron. No PVH-projecting ARCOxtr neurons are GABAergic. b, Expression of AAV-DIO-hM3Dq-mCherry in ARCOxtr neurons as determined by mCherry immunolabeling. c, Expression of AAV-DIO-hM3Dq-mCherry (left) and Fos (middle) immunolabeling in ARCOxtr neurons (right, overlay) in mice sacrificed 90 minutes after CNO administration. d, CNO/hM3Dq stimulation of ARCOxtr reduces refeeding following 24 hours fasting. Mice from multiple litters, n = 5 animals. Repeated measures two-way ANOVA: Treatment F(1,4) = 10.91, P = 0.0298; time F(4,16) = 64.22, P < 0.0001; interaction F(4, 16) = 6.976, P = 0.0019. Sidaks multiple comparisons test: 1h, P = 0.2882; 2h, **P = 0.0018; 3h, ****P = 0.0003; 4h, ****P < 0.0001. Data are presented as mean ± s.e.m. e, Expression of AAV-DIO-ChR2-mCherry (left) in ARCOxtr neurons and placement of optical fiber above the PVH (right).
