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. 2020 May 8;11:2261. doi: 10.1038/s41467-020-16203-x

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© The Author(s) 2020

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. 2 — a Retrograde labelling by CTB-488 microinjection in the LSv resulted in prominent expressions in the ventral subiculum (VS) of the hippocampal formation in the ipsilateral brain. Scale bar, 1000 μm. Inset shows enlarged ventral subiculum. Scale bar, 200 μm. b Anterograde tracing by injection of AAV9-CamIIKα-ChR2-mCherry confirmed the VS-LSv projection. Left, scale bar, 1000 μm; Right, scale bar, 200 μm. c Anterograde tracing by AAV9-hSyn-hChR2(H134R)-eYFP microinjection in the LSv resulted in prominent expressions in the tuberal nucleus (Tu) of the lateral hypothalamus. Left, scale bar, 1000 μm; Right, scale bar, 200 μm. d Injection of CTB-488 into Tu confirmed the LSv-Tu projection. Left, scale bar, 1000 μm; middle, scale bar 500 μm; Right, scale bar 200 μm. e and f Optogenetic activation of the VS → LSv pathway for 5 min at 25 Hz induced increased grooming behaviour that was significantly higher than those in the 5-min pre-light and post-light-off periods (n = 5, One-way repeated measures ANOVA with Tukey post-hoc test, F (1.489, 5.954) = 13.05, P = 0.0083; off vs. on, *P = 0.0189; on vs. post-off, **P = 0.0096). g Comparison of the delay and time spent in grooming and arousal behaviours, including rearing and heading, in the initial 2 min of stimulation. n = 5; delay time, ***P = 0.0004; time spent, ***P = 0.0009; Student’s paired two-tailed t-test. h and i Optogenetic activation of the LSv → Tu pathway for 5 min induced increased grooming behaviour that was significantly higher than those in the 5-min pre-light-off and post-light-off periods. (n = 7, one-way repeated measures ANOVA with Tukey post-hoc test, F (1.104, 6.622) = 39.73, P = 0.0004; pre-off vs. on, **P = 0.0025; on vs. post-off, ***P = 0.0010). j Comparison of the delay and time spent in grooming and arousal behaviours, including rearing and heading, in the initial 2 min of stimulation. (n = 7; delay time, ***P = 0.0002; time spent, **P = 0.0147; Student’s paired two-tailed t-test). All data are are presented as mean ± SEM. Source data are provided as a Source Data file.