Fig. 7

Effects of PAE on oxidative DNA damage in frontal cortex and hypothalamus of rat brain. Frozen sections from frontal cortex (a) and hypothalami (b) of 6-day-old rat pups were immunostained for γH2Ax (red) and foci were counted using ImageJ (National Institutes of Health, Bethesda, Maryland). Boxed areas in DAPI stained sections indicate the area where representative photographs of prefrontal cortex and hypothalamus were taken. A higher magnification of γH2Ax Foci is shown in the insets. Data are shown as mean ± SE, n = 4–6 animals per group done in duplicate. 3V, third ventricle in the hypothalamus. Stains for γH2Ax are shown in red and DAPI in blue. Extracted and hydrolyzed DNA from frontal cortex and hypothalamus (c) of 6-days-old PF and AF rat pups was used to confirm the PAE-mediated oxidative DNA damage by measuring 8-hydroxy 2 deoxyguanosine (8-OHdG) using a competitive 8-OHdG ELISA, as described in the “Methods” section. Effects of the free radical spin trap PBN (α-phenyl-N-tert-butyl nitrone) on 8-OHdG formation in AF compared to PF animals was measured in prefrontal cortex (d). Data are shown as mean ± SE, n = 5–8 done in triplicate