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. 2018 Aug 7;9:3133. doi: 10.1038/s41467-018-05565-y

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© The Author(s) 2018

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 — DCN neuron activity is required for goal-directed behaviors. a Stereotaxic injection and analysis scheme. b Left: Example image of AAV-DiO-hM4Di infection site in DCN of vGluT2-cre mice. Right: Quantification of hM4Di-positive neurons in each DCN subnucleus (N = 10 mice, mean ± SEM). Scale bar: 400 µm. c Open field tracks of control DCN^GFP and hM4Di-expressing DCN^hM4Di treated CNO. Right: Quantification of distance traveled in open field and velocity. (N = 11 and 10 mice, respectively; mean±SEM, unpaired t-test). d Left: example images of marble burying test. Right: number of marbles buried by DCN^GFP and DCN^hM4Di mice treated with CNO (N = 9 mice per group, mean ± SEM, unpaired t-test). e Left: Cartoon for T-maze test. Right: Number of spontaneous alternations observed in 10 consecutive trials in a T-maze without food reward. (N = 10 mice per group, mean ± SEM, unpaired t-test). f T-maze reversal learning. Mice were food-deprived and trained in a rewarded T-maze paradigm to choose one arm. Percentage of mice reaching the training criterion after each day is plotted (N = 9 DCN^GFP and N = 8 DCN^hM4Di mice). g T-maze reversal learning. Percentage of correct choice at Day1 and Day 2 after reversal of the rewarded side. For each day, the first 5 and second 5 trials are plotted separately to differentiate perseverative errors (p = 0.325, unpaired t-test) and regressive errors (p = 0.347, unpaired t-test; N = 9 DCN^GFP and N = 8 DCN^hM4Di mice, mean ± SEM). h T-maze forced alternation. The percentage of mice reaching the training criterion after each day is plotted. (N = 10 DCN^GFP and N = 9 DCN^hM4Di mice). i T-maze forced alternation. Performances at the last training session (-CNO) and the session after CNO treatment (+CNO) are compared. For comparison between DCN^GFP and DCN^hM4Di mice after CNO, mean ± SEM, Mann–Whitney test, two tailed. For comparison of DCN^hM4Di mice before and after CNO, mean ± SEM, Mann–Whitney test, two tailed (N = 10 DCN^GFP and N = 9 DCN^hM4Di mice). j Correlation of the percentage of hM4Di-positive neurons in IntP and the performance in the forced alternation task after CNO treatment in DCN^hM4Di mice. Each circle represents one animal. Thick-lined circles represent mice with >50% of IntP neurons expressing hM4Di. Thin-lined circles are mice not included (<50% of IntP neurons, N = 8 mice)