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. 2019 Jun 25;9:9225. doi: 10.1038/s41598-019-45667-1

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© The Author(s) 2019

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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Prenatal valproate exposure induced reductions in cerebellar Purkinje cell number. (A) Light micrograph of calbindin D28k (CB) immunostained sagittal section of rat cerebellar vermis at low magnification. (B) Boxplots representing normalized CB immunoreactive Purkinje cell numbers in cerebellar vermal lobules 6a, 6b, 6c and 7, for the VEH, VPA400 and VPA600 groups. Asterisks depict significant (p < 0.05) decreases compared to the VEH group. (C,D) Normal distribution of CB immunoreactive Purkinje cells in the ganglionic layer of cerebellar cortex on VEH sections. (E–H) Decreased CB-positive Purkinje cell number, found both in the VPA400 and VPA600 groups. The arrows point to unlikely large areas with lack of Purkinje cells. Scale bars: 1000 μm (A), 100 μm (C,E,G), 20 μm (D,F,H).