Fig. 2

Hyperammonemia increases the content of IL-17 receptor in microglia and its membrane expression in cerebellum. Blocking IL-17 with anti-IL-17 reverses these effects. Membrane expression of the IL-17 receptor (G) was analyzed using BS3 cross-linker in slices from control (C) and hyperammonemic (HA) rats. The effects of hyperammonemia and of blocking IL-17 with anti-IL-17 in white matter (A) and Purkinje neurons (B) were analyzed by immunohistochemistry with DAB staining using antibodies against IL-17 receptor. Representative images are shown in A-B. The content of IL-17 receptor in Purkinje neurons and granular neurons was negligible. Double immunofluorescence staining was performed using anti-IL-17 receptor and anti-IBA1 (C, E) or GFAP (D, F). Representative images of the double fluorescence staining are shown (C, D). Values are mean ± SEM of 10–20 rats per group in G and 4–6 rats per group in A–D. One-way ANOVA followed by Fisher’s LSD post-hoc test was performed to compare all groups. Values are the mean ± SEM. Values significantly different from control group are indicated by asterisk (*p < 0.05) and values significantly different from HA group are indicated by a (a = p < 0.05, aa = p < 0.01). Yellow arrows indicate co-localization of the two proteins while white arrows indicate lack of co-localization. As the IL-17 receptor is expressed both in astrocytes and microglia, there are some instances of double immunofluorescence in which the markers (Iba1 or GFAP) and the IL-17 receptor do not co-localize effectively. We indicate this situation with white arrows. For example, in C there are some cells that are stained by anti-IL-17R but not by anti-Iba1. These cells may be for example astrocytes, which also express IL-17R. The same occurs in D, some cells (likely microglia) are stained by anti-IL-17R but not by anti-GFAP