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. 2018 Jan 4;9:52. doi: 10.1038/s41467-017-02534-9

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© The Author(s) 2017

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. 3 — LH^Pdx1→PVH evoked grooming requires glutamate release, and activation of non-glutamatergic fibers promotes behavioral approach. Food intake (a) and grooming time (b) before (pre-light), during (light-on), and after (post-light) 5 Hz, 100 ms photostimulation of LH^Pdx1-ChR2→PVH fibers in Pdx1-Cre (n = 8) and Pdx1-Cre::Vglut2 ^flox/flox (n = 7) mice. Real-time place preference assay (RTPP) during optical stimulation of LH^Pdx1→PVH in GFP control (n = 6) and Pdx1-Cre::Vglut2 ^flox/flox::ChR2 mice (n = 4). c, d Representative tracks tracing locomotion; e Percentage time spent on each side. e Two-way ANOVA followed by Sidak’s multiple comparisons test (interaction F [1, 16] = 7.192, P = 0.0164; viral injection F [1, 16] = 0.1407, P = 0.7125; light zone F [1, 16] = 6.612, P = 0.0205. Pdx1-Cre::Vglut2 ^flox/flox::ChR2 (Light-off zone vs. Light-on zone) **p < 0.005. Data presented as mean ± s.e.m. Food intake (f) and grooming time (g) before, during, and after 5 Hz, 10 ms photostimulation of LH^Pdx1-ChR2→PVH fibers in Pdx1-Cre (n = 4) and Pdx1-Cre::Vglut2 ^flox/flox (n = 3) mice. a, b, f and g Two-way repeated measures ANOVA followed by Dunnett’s multiple comparisons test. (a interaction F [2, 26] = 0.003466, P = 0.9965; genotype F [1, 13] = 0.000817, P = 0.9776; light F [2, 26] = 66.15, P < 0.0001, subjects (matching) F [13, 26] = 0.9467, P = 0.5233. Pdx1-Cre and Pdx1-Cre::Vglut2 ^flox/flox [pre-light vs. light-on] ***p < 0.0005. b interaction F [2, 26] = 1.065, P = 0.3593; genotype F [1, 13] = 10.98, P = 0.0056; light F [2, 26] = 3.622, P = 0.0409; subjects (matching) F [13, 26] = 1.472, P = 0.1942. f interaction F [2, 10] = 3.073, P = 0.0912; genotype F [1, 5] = 3.073, P = 0.1400; light F [2, 10] = 3.808, P = 0.0590; subjects (matching) F [5, 10] = 1, P = 0.4651. Pdx1-Cre::Vglut2 ^flox/flox (pre-light vs. light-on) *p < 0.05. g interaction F [2, 10] = 14.45, P = 0.0011; genotype F [1, 5] = 8.36, P = 0.0341; light F [2, 10] = 8.57, P = 0.0068; subjects [matching] F [5, 10] = 1.078, P = 0.4280. Pdx1-Cre (pre-light vs. light-on) ***p < 0.0005)