Extended Data Fig. 5: Gi-DREADD-driven inhibition of acclimation-induced VMPOLepR activity prevents heat tolerance.
From: Thermally induced neuronal plasticity in the hypothalamus mediates heat tolerance
a, Schematic drawings representing different anatomical positions along the rostral caudal axis of the preoptic hypothalamic region with the 3 middle drawings (approx. bregma = 0.5 mm to bregma = 014) indicating the center of the VMPO region (top) with corresponding typical fluorescent images depicting the extent of virally (AAV) delivered Cre-dependent Gi-DREADD expression in a LepR-Cre mouse (bottom). b, Top: Schematic showing the protocol used for ex vivo verification of CNO triggered, Gi-DREADD mediated inhibition of VMPOLepR following heat acclimation. Bottom left: Representative electrophysiological traces showing the effect of CNO on the firing pattern of acclimated VMPOLepR neurons injected with either Cre-dependent Gi-DREADD-mCherry AAV or only a Cre-dependent mCherry control AAV. Bottom right: average (mean ± s.e.m.) tonic AP firing frequency of acclimation-induced VMPOLepR cells in the presence of 5 µM CNO. Mann-Whitney U test (one-tailed), ***P < 0.0001. n = 35/3 cells per group. c, Heat endurance assay: Average (mean ± s.e.m.) body temperature of non-acclimated (top) or acclimated (bottom) Gi-DREADD-positive and CNO-injected animals during the heat endurance assay. The same animals injected with saline instead of CNO were also plotted for comparison. N = 8 mice for the non-acclimated and N = 7 mice for the acclimated condition.
