Extended Data Fig. 3: lPBN Y1R neuron ablation suppresses pain-related behaviors without affecting locomotion or responses to other negative stimuli.
From: A parabrachial hub for need-state control of enduring pain
(a) Viral injection strategy for bilateral lPBN Y1R neuron ablation. (b) Experimental timeline for lPBN Y1R neuron ablation. (c) Representative images showing Npy1r mRNA in the lPBN labeled using RNAscope fluorescence in situ hybridization from a control and DTR-expressing mouse. Scale bar, 200 μm; insets, 50 μm. (d) Number of cells / section expressing Npy1r in the lPBN in control (n = 4) and ablated (n = 5) mice (unpaired two-sided t-test, p = 0.0023). (e) The formalin test was used to test the effect of lPBN Y1R neuron ablation on pain behavior. (f) Time spent licking paw in 5 min bins after an injection of formalin in control or Y1R ablated mice (n = 8 mice/group, two-way ANOVA, main effect of group p = 0.0205, group x time interaction p = 0.0217). (g) Time spent licking paw during phase 1 in (f) (n = 8 mice/group, unpaired t-test, p = 0.0042). (h) Time spent licking paw during phase 2 in (f) (n = 8 mice/group, unpaired t-test, p = 0.03). (i) Representative plots showing locomotion of control and ablated mice. (j-l) Distance traveled (j), time spent immobile (k), and time spent in the center of the arena (l) in control (n = 6) and ablated (n = 5) mice (unpaired t-tests, n.s.). (m) Conditioned taste aversion paradigm. (n-o) Amount of gel paired with saline or LiCl consumed by control and Y1R-ablated mice after 30 min (n) and 60 mins (o) (n = 6 control vehicle, n = 5 control ablated, n = 8 LiCl mice, two-way ANOVA, main effect of virus n.s.). (p) Predator odor avoidance paradigm. (q) Number of entries into the odor zone before and after addition of TMT in control (n = 6) and Y1R-ablated (n = 5) mice (two-way ANOVA, main effect of virus n.s.). (r) Time spent in the odor zone before and after addition of TMT in control (n = 6) and Y1R-ablated (n = 5) mice (two-way ANOVA, main effect of virus n.s.). Data are expressed as mean ± SEM. Grey dots represent individual mice. T-test and post-hoc comparisons: *p < 0.05, **p < 0.01, ***p < 0.001. ANOVA main effect of group: #p < 0.05. ANOVA interaction: †p < 0.05.
