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. 2017 Aug 11;7:7986. doi: 10.1038/s41598-017-08208-2

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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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Comparison of neuronal and glial cell alterations in the ACC. Results of double staining of rat brain slices for the astrocyte marker GFAP and neuron marker NeuN are shown. DAPI counterstaining is indicated by blue fluorescence. (A–D) The expression of GFAP and NeuN in neuronal cells in sham NP rats is shown. (E–U) GFAP immunoreactivity in the ACC was elevated in all experimental groups. However, the expression levels of NeuN did not significantly change in all animal models used (scale bar, 50 μm). (V) Western blot analysis was used to detect GFAP and NeuN protein levels in the ACC. GFAP expression in the ACC significantly increased in the NP + Sham MCS, NP + MCS, NP + MCS + ZIP, and NP + MCS + Vehicle rats. Elevated NeuN expression was detected in NP + MCS and NP + MCS + ZIP rats, but significant changes were not observed. β-Actin was used as a loading control. *P < 0.05 compared with sham NP and n = 4 rats for each group.