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. 2019 Nov 22;9:17402. doi: 10.1038/s41598-019-53192-4

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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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KA administration induces loss of large number of GABA cells but GABAergic Shh-nlacZ+ neurons seem unaffected. (A–D) Shh-nlacZ+ GAD 65/67+ neurons in the DG of control animals showing co-localization (25X, bar 100 µm). Note the full complement of cells expressing Shh-lacZ in the hilus (red channel). (E–H) The number of Shh-nlacZ+ neurons did not decrease while the number of GAD 65/67+ neurons appears to diminish at 2 weeks following KA injections (25X, bar 100 µm). (J) Cells expressing Shh-nlacZ were up-regulated, while the overall expression of GAD 65/67 decreased. GABAergic subtypes included PV, parvalbumin; SST, somatostatin; CR, calretinin and Shh, sonic hedgehog. Examples of these staining are provided in Fig. 2. (p values correspond to Unpaired Student’s t-Test). (K–N) Behavior was analyzed on the second week after injection, scores correspond to average counts/h or seconds/h of a given behavior for 7 days, 4 h/day(12–4 pm) and 4 h/night (12–4am). *p < 0.01, **p < 0.001, night vs. day; & p < 0.001 KA vs. vehicle group. Epileptiform activity was always equal or lower than stage 3 (mouse forelimb clonus without rearing) (n = 6/group). Abbr: GCL, granular cell layer; SGZ subgranular zone; H, hilus; M.A., motor activity.