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. 2022 Nov 11;13:6865. doi: 10.1038/s41467-022-34517-w

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© The Author(s) 2022, corrected publication 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 6 — a Surgical strategy for optogenetic manipulation of PVT→NAc neurons and intra-NAc DAMGO infusions. b–d Intra-NAc infusion of DAMGO prevented the suppression of sucrose self-administration caused by PVT→NAc stimulation (b), TMT (c), and yohimbine (d) (Opto: n = 8 mice/group; one-way ANOVA, F_2,21 = 9.33, P = 0.001; planned two-tailed t-tests: base vs. opto P < 0.01, opto vs. DAMGO + opto P < 0.01; TMT: n = 8 mice/group; one-way ANOVA, F_2,21 = 8.02, P = 0.003; planned t-tests: base vs. TMT P < 0.01, TMT vs. DAMGO + TMT P = 0.02; yohimbine: n = 7 mice/group; one-way ANOVA, F_2,18 = 4.26, P = 0.03; planned t-tests: base vs. yohimbine P = 0.02, yohimbine vs. DAMGO + yohimbine P < 0.05). e Surgical strategy for patch-clamp electrophysiology. f Example electrophysiological waveforms (scale: 50pA/50 ms) and grouped data reveal that DAMGO reduced PVT→NAc^MSN and PVT→NAc^PV-IN oeEPSC amplitudes (MSNs: n = 10 cells, 6 mice; PV-INs: n = 8 cells, 5 mice; repeated-measures two-way ANOVA, effect of DAMGO: F_1,16 = 210.9, P < 0.0001; Sidak’s post-hoc: P-values < 0.001). g Surgical strategy for knockout of PVT µ-ORs with patch-clamp electrophysiology. h Example electrophysiological waveforms (scale: 50pA/50 ms) and grouped data reveal that PVT µ-OR knockout prevented the DAMGO-induced decrease in PVT→NAc^MSN and PVT→NAc^PV-IN oeEPSC amplitudes (n = 4 cells per group, 2 mice; repeated-measures two-way ANOVA, effect of DAMGO F_1,6 = 0.0001 P = 0.99). i Surgical strategy for PVT µ-OR knockout with simultaneous optogenetic manipulation of PVT→NAc neurons and intra-NAc DAMGO infusions. j–l Knockout of PVT µ-ORs in Oprm1^fl/fl mice rescued the suppression of sucrose self-administration caused by optogenetic stimulation of PVT→NAc neurons (j), TMT (k), and yohimbine (l) despite intra-NAc DAMGO infusions (Opto: n = 6 Oprm1^fl/fl, 8 WT mice; repeated-measures two-way ANOVA, day × group interaction: F_2,24 = 9.74, P = 0.001; Sidak’s post-hoc: opto + DAMGO P = 0.02; TMT: n = 6 Oprm1^fl/fl, 8 WT mice; repeated-measures two-way ANOVA, day × group interaction: F_2,24 = 7.04, P = 0.004; Sidak’s post-hoc: TMT + DAMGO P < 0.001; yohimbine: n = 6 Oprm1^fl/fl, 8 WT mice; repeated-measures two-way ANOVA, day × group interaction: F_2,24 = 10.97, P = 0.0004; Sidak’s post-hoc: yohimbine + DAMGO P < 0.05). µ-OR µ-opioid receptor, Base Baseline, Opto optogenetics, Yoh Yohimbine, KO knockout; Group comparisons: *P < 0.05, **P < 0.01, ****P < 0.001. Bar graphs are presented as mean values ± SEM. Source data are provided as a Source Data file.