Figure 4: Expression of mGluR5 mRNA (a, c, e, and g) and protein (b, d, f, and h) in forebrain of control rats and rats with EAE at different times post-immunization (a and b) and after therapeutic treatment with antagonists of glutamate receptors: amantadine (c and d), memantine (e and f), and MPEP (g and h). Total RNA was prepared from healthy control rats, rats with EAE, and rats with EAE after therapy at the indicated d.p.i. The mGluR5 mRNA levels were determined by quantitative real-time PCR (see Section 2) and normalized to actin. Graphs (a), (c), (e), and (g) present the results expressed as percentage of control from four independent experiments. ; ; significantly different versus control (healthy untreated rats). ; ; significantly different versus rats with EAE not subjected to therapy at the corresponding d.p.i. (one-way ANOVA followed by Dunnett’s multiple comparison posttest). Representative immunoblots show the expression of mGluR5 receptor protein in forebrain homogenates of (b) control rats and rats with EAE at different times post-immunization, (d) amantadine-treated rats with EAE, (f) memantine-treated rats with EAE, and (h) MPEP-treated rats with EAE. The results are expressed as percentage of control. Graphs (b), (d), (f), and (h) present the results of densitometric analysis, normalized to β-actin, of four independent immunoblots, each done from distinct brain; (one-way ANOVA with post hoc Dunnett’s test).