Journal: Translational psychiatry
Article Title: Maternal immune activation results in complex microglial transcriptome signature in the adult offspring that is reversed by minocycline treatment.
doi: 10.1038/tp.2017.80
Figure Lengend Snippet: Figure 4. Signs of classical pro-inflammatory activation in Poly(I:C) hippocampal microglia. (a) Representative image of a sagittal slice (from a control animal) used for the analysis of the binding capacity with the TSPO-specific radioligand [18F]GE180. (b) Shows a representative image of a Nissl-stained sagittal slice (control animal) used to localize specific brain regions. (c) Shows that Poly(I:C) mice exhibit an increased binding potential to the TSPO in the hippocampus (n = 5 mice) with respect to controls (n = 6 mice). On the other hand, slices from Poly(I:C) mice treated with minocycline display a normalized binding potential in the latter region (n = 6 mice), significantly lower than untreated Poly(I:C) animals and comparable to controls. (d) Representative pictures illustrating increased Iba1 immunoreactivity in the proximity of the DG of Poly(I:C) mice as compared with controls and minocycline-treated mice. Pictures were taken at a 63-times magnification. (e) Iba1 immunoreactivity was increased in the dentate gyrus of the hippocampus (DG) of Poly(I:C) mice (n = 7) as compared with controls (n = 5). Poly (I:C) animals treated with minocycline (n = 4) displayed a normal Iba1 immunoreactivity. (f–h) Enzyme-linked immunosorbent assay (ELISA) measurement in whole hippocampal homogenates of the pro-inflammatory cytokines interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α). Only IL-6 was found to be significantly increased (h), while the levels of the other cytokines remained unchanged (f and g; Controls, n = 5; Poly(I:C), n = 5; Poly(I:C) treated with minocycline, n = 4). Error bars represent s.e.m. in all the panels. The data from the radioligand-binding assay were analyzed by one-way analysis of variance (ANOVA) followed by Bonferroni post hoc test. The data from the immunoreactivity and ELISA were analyzed by one-way ANOVA followed by Newman–Keuls post hoc test. *Po0.05, **Po0.001. Cb, cerebellum; Cc, corpus callosum; ctrl., control animals; Ctx, cortex; Hip, hippocampus; Ob, olfactory bulbs; Poly, Poly(I:C) animals; Poly/Mino, Poly(I:C) animals treated with minocycline; Th, thalamus; TSPO, translocator protein.
Article Snippet: Complement component 4a (C4a) levels in hippocampal lysates were measured using the mouse C4a enzyme-linked immunosorbent assay (ELISA) kit (Elabscience Biotechnology, Wuhan, China), according to the manufacturer’s manual.
Techniques: Activation Assay, Control, Binding Assay, Staining, Enzyme-linked Immunosorbent Assay, Radio Ligand Binding Assay
![Figure 4. Signs of classical pro-inflammatory activation in Poly(I:C) hippocampal microglia. (a) Representative image of a sagittal slice (from a control animal) used for the analysis of the binding capacity with the TSPO-specific radioligand [18F]GE180. (b) Shows a representative image of a Nissl-stained sagittal slice (control animal) used to localize specific brain regions. (c) Shows that Poly(I:C) mice exhibit an increased binding potential to the TSPO in the hippocampus (n = 5 mice) with respect to controls (n = 6 mice). On the other hand, slices from Poly(I:C) mice treated with minocycline display a normalized binding potential in the latter region (n = 6 mice), significantly lower than untreated Poly(I:C) animals and comparable to controls. (d) Representative pictures illustrating increased Iba1 immunoreactivity in the proximity of the DG of Poly(I:C) mice as compared with controls and minocycline-treated mice. Pictures were taken at a 63-times magnification. (e) Iba1 immunoreactivity was increased in the dentate gyrus of the hippocampus (DG) of Poly(I:C) mice (n = 7) as compared with controls (n = 5). Poly (I:C) animals treated with minocycline (n = 4) displayed a normal Iba1 immunoreactivity. (f–h) Enzyme-linked <t>immunosorbent</t> assay <t>(ELISA)</t> measurement in whole hippocampal homogenates of the pro-inflammatory cytokines interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α). Only IL-6 was found to be significantly increased (h), while the levels of the other cytokines remained unchanged (f and g; Controls, n = 5; Poly(I:C), n = 5; Poly(I:C) treated with minocycline, n = 4). Error bars represent s.e.m. in all the panels. The data from the radioligand-binding assay were analyzed by one-way analysis of variance (ANOVA) followed by Bonferroni post hoc test. The data from the immunoreactivity and ELISA were analyzed by one-way ANOVA followed by Newman–Keuls post hoc test. *Po0.05, **Po0.001. Cb, cerebellum; Cc, corpus callosum; ctrl., control animals; Ctx, cortex; Hip, hippocampus; Ob, olfactory bulbs; Poly, Poly(I:C) animals; Poly/Mino, Poly(I:C) animals treated with minocycline; Th, thalamus; TSPO, translocator protein.](https://pub-med-unpaywalled-images-cdn.bioz.com/pub_med_ids_ending_with_5733/pm28485733/pm28485733__page8_image1.jpg)
