Fig. 4

Correction by LGZ of the bioenergetic component in hippocampus and cerebellum, together with a phenotypic amelioration in symptomatic Rett mice. A Treatment with LGZ was assayed in Rett mice, treating them from weaning until sacrifice at the symptomatic stage (7 m.o.). Both ATP concentration (detected by bioluminescence) and lipid peroxidation (detected by TBARS) were assayed in untreated and LGZ-treated Rett mice in B cortex, C hippocampus and D cerebellum. E to G show results regarding oxidative stress and antioxidant markers in the brain areas of interest. E Detection of the RNA oxidative damage marker 8-OHdG in cerebellum by immunofluorescence. 8-OHdG staining is shown in magenta and calbindin in yellow for Purkinje cells in the upper images at 10× magnification; in the lower images 8-OHdG is shown with the intensity-revealing LUT “Green Fire Blue”, with a 2× electronic zoom; scale bar represents 50 µm in both images. F Antioxidant markers MnSOD and GPX in cortex, hippocampus and cerebellum detected by western blot, and their respective densitometry quantification, normalized by tubulin (G). Phenotypical characterization of the treated mice, registering an improvement in the General Health Score (H), which was maintained until the endpoint (I) and a recovery of the explorative behaviour of Rett mice upon treatment in the NOR-test (J). All experiments were done in at least three different mice, with three technical replicates and each experiment was done in triplicated (except for lipid peroxidation). Statistical analysis was performed as described in Materials and Methods. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Absence of asterisk means no statistically significant difference. Wild-type female mice in grey; Rett female mice in yellow; LGZ-treated Rett female mice in green