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. 2020 Jul 6;10:11047. doi: 10.1038/s41598-020-68110-2

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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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Activation of GPR40 in synergy with BDNF and interleukin-6 increases neuronal differentiation of hypothalamic neural precursor cells. Neurospheres were prepared from hypothalamic NPC of postnatal day 1 C57BL/6J mice and treated with TUG905, BDNF or Il-6 during 7 or 18 days of differentiation (A). Immunostaining of DCX/GFAP positive cells in 7 days differentiated cultures and of MAP2/GFAP positive cells in 18 days differentiated cultures (B). Gene expression analysis of 7 days differentiated NPC (C, D) identified increased mRNA levels of DCX in TUG905, BDNF and Il-6 treated cells (D) as well as a higher number of DCX-positive neuroblasts (E). In 18 days differentiated NPC there was no difference on gene expression of GPR40, GFAP or hypothalamic neuropeptides after TUG905, BDNF or Il-6 treatments (F–J), despite the increase on MAP2-positive mature neurons (K). Scale bar 25 μm. Data are presented as means ± SEM. N = 3–4 per group per group. *p < 0.05, one-way ANOVA followed by Tukey’s post hoc test.