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. 2019 Jul 10;13:306. doi: 10.3389/fncel.2019.00306

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Copyright © 2019 Tenza-Ferrer, Magno, Romano-Silva, da Silva and Gomez.

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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Peripheral inflammation induces microglia reactivity in the spinal cord. (A) Representative fluorescent images and (B) quantification of Iba1-labeled microglia (Iba1⁺) in the ipsilateral dorsal horn of the lumbar spinal cord, showing the saline and CFA groups in 2, 7, or 14 days after the paw injection. All values are relative to each respective saline group (n = 3 for saline and n = 4 for CFA groups, six slices per animal). (C) Representative fluorescent images of Iba1-labeled microglia in the dorsal horn of the spinal cord, showing the contralateral (left) and ipsilateral (right) sides of the saline (Stevens et al., 2007) and CFA (bottom) groups. (D) Left: Intragroup analysis of the number of Iba1⁺ cells of the ipsilateral relative to the contralateral side in each group (saline in blue, CFA in red). Right: Intergroup analysis of the number of Iba1⁺ cells of the contralateral and ipsilateral sides of CFA-treated relative to the saline-injected rats (contralateral in purple, ipsilateral in gray; n = 4, six slices per animal; #### p < 0.0001). (E) Representative images of the microglia morphology in the ipsilateral saline (Stevens et al., 2007) and CFA (bottom) groups in 2 (left) or 7 (right) days after the paw injection. Left panel: Iba1+ immunofluorescence; Middle panel: the binary transformation; Right panel: the resulting skeleton. The insets represent the respective higher magnification image. (F,G) Quantification of the microglia morphology parameters in ipsilateral side of 2 (F) or 7 (G) days after the paw injection. The graphs show the quantification of the number of branches (Stevens et al., 2007) and junctions (middle), or the total branches length (bottom) per microglia (n = 3–4 rats/group, three slices per animal). Each point in the graphs represents one microglia. Scale bars are 100 μm (A,C), 50 μm (E) or 10 μm (E, insets). M: medial; L: lateral; D: dorsal; V: ventral (coordinates relative to the central canal). Summary data are represented as mean ± SEM. ^∗p < 0.05; ^∗∗p < 0.01; ^∗∗∗p < 0.001; ^∗∗∗∗p < 0.0001; n.s., not significant.