Figure 5
Overexpression of KCNQ3 in the LHb of EtOH-WD rats attenuated thermal hyperalgesia and ethanol consumption. (A) KCNQ3-YFP expression after viral vector injection and immunofluorescence of the neuronal marker, NeuN. A Strong signal of KCNQ3-YFP overlaps with NeuN. Scale bar: 10 μm. (B) Example and pooled results of Western blots show the increased KCNQ3 expression in the LHb from EtOH-WD rats injected with KCNQ3-eYFP viruses compared to that injected with eYFP control viruses. Unpaired t-test, t = 3.18, *p < 0.05, Nrat = 4/group. Full-length blots are presented in Supplementary Fig. 1. (C) CCD camera captured IR (left) and ET-DSRed filtered fluorescence (right) image of the LHb neuron after viral injection for patch-clamp recording. (D,E) Sample traces (D) and pooled data (E) showing increased inward current relaxations in neurons of EtOH-WD rats after KCNQ3-eYFP virus injection. Two-way ANOVA, for YFP/KCNQ F1,17 = 5.485, P = 0.0316, post-hoc: **p < 0.01. Ncell = 9–10/group. Scale bar: 200 pA and 200 ms. (F) Paw withdrawal latency was significantly increased after KCNQ3 overexpression in the LHb. Two-way ANOVA, for Before/After F1,16 = 9.211, P = 0.0079, for Interaction F1,16 = 16.06, P = 0.0010, post-hoc: ***p < 0.001 vs. Before, #p < 0.05 vs. YFP, Nrat = 8–10/group. (G) KCNQ3-eYFP-overexpressing rats showed reduced ethanol intake relative to eYFP-overexpressing rats. Unpaired t-test, t = 2.342, *p < 0.05. Nrat = 8–9/group.
