Figure 6.

Overt oxidative stress and mitochondrial deficits occurring in brains from ICV-STZ mice are recovered by 12A12mAb immunization. (A–I) Cortical homogenates from animals of four experimental groups (STZ 0 plus vehicle; STZ 0 plus mAb; STZ 3 plus vehicle; STZ 3 plus mAb) were assessed for oxidative stress and oxidative phosphorylation. In (A,B): ROS level. Cytosolic and mitochondrial O2^−• productions were detected according to the adrenochrome method (A) and by using the MitoSOX dye (B), respectively. The O2^−• level value was expressed as percentage of STZ 0 plus vehicle, to which value 100 was given. In C–D: Lipid peroxidation and COX activity. Index of lipid peroxidation (C) are expressed as fluorescence values at the beginning of the reaction (t0) minus the value measured after 30 min (t30), accordingly to [66]. COX activity (D) was determined by measuring polarographically O2 ^{− •} consumption in the oxygen electrode chamber [74]. In (E–G): GSH, NADH and NADPH level determinations were calculated as in [68]. In (H): MRC complex activities: the activities of Complex I (NADH:ubiquinone oxidoreductase), Complex II (succinate:ubiquinone oxidoreductase), Complex III (cytochrome c reductase), Complex IV (cytochrome c oxidase) and Complex V (ATP synthase) [39]. In (I): Cellular ATP content [39]. Values are from at least three independent experiments and statistically significant differences were calculated by one-way ANOVA followed by Bonferroni’s post-hoc test for multiple comparison among more than two groups. p < 0.05 was accepted as statistically significant (** p < 0.01; *** p < 0.0005; **** p < 0.0001).