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. 2018 Dec 10;12:479. doi: 10.3389/fncel.2018.00479

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Copyright © 2018 Sancandi, Schul, Economides, Constanti and Mercer.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Neuroinflammation in the PC of the pre-motor PD model. (A) Astrocytic activation in the pre-motor PD model. (Aa) Representative immunohistochemical staining of GFAP in a sham animal, model and model treated with EX-4. Scale bars for overview and for inserts represent 200 and 50 μm, respectively. (Ab) Thresholding analysis of GFAP staining revealed an increase in the intensity in the pre-motor model compared with that in sham animals suggesting an activation of astrocytes. This increase was prevented by treatment with EX-4 (unpaired t-test, ^∗∗∗∗P < 0.0001). Data are represented as mean ± SEM. (B) Microglia activation in the pre-motor PD model. (Ba) Representative immunohistochemical staining of Iba1 in a sham animal, model and model treated with EX-4. High magnification image of one Iba1-positive cell for each condition is represented in the inserts. Scale bars for overview and for inserts represent 200 and 10 μm, respectively. (Bb) Thresholding analysis of Iba1 staining revealed a decrease in the intensity in the pre-motor model compared with that in sham animals suggesting an activation of microglia. This decrease was not prevented by treatment with EX-4 (unpaired t-test, ^∗P_shamvs. _model ₊ _EX-4 <0.05, P_{modelvs.model} ₊ _EX-4 >0.05). Data are represented as mean ± SEM. (C) Morphological characteristics of Iba1-immunopositive cells. (Ca) Neurolucida reconstructions of Iba1-positive cells in the sham, model and model + EX-4. Each microglia branch is represented with a different color. Scale bars represent 20 μm. (Cb) Number of intersections between processes of Iba1-positive cells and concentric sphere at different radius from the soma in the sham, model and model treated with EX-4 (Sholl analysis). Data are represented as mean ± SD (n = 8 per experimental group – unpaired t-test – ^∗∗P < 0.01 for pre-motor model compared with sham; && P < 0.01 for sham compared with pre-motor model treated with EX-4). (Cc–g) The number of branches (Cc), total length (Cd), total surface (Ce), and the number of nodes (Cf) of Iba1-positive cells were decreased in the pre-motor PD model compared with those in the sham animals. Treatment with EX-4 did not prevent this decrease. The size of cell somata was similar in the sham, model and model + EX-4 (Cg). Data are represented as mean ± SD (n = 8 per experimental group – unpaired t-test ^∗P < 0.05, ^∗∗P < 0.01).