Figure 5

Apoptosis in primary injury phase. Representative examples of high magnification (63X, zoom 2) confocal images of Neurons (NeuN, A and B) or oligodendrocytes (OLIG1, C and D) (green), Caspase-3 (red) and their colabeling in ventral horns (VH) of injured spinal cord from saline- (SAL) or DHA-treated animals, 7 days after SCI, at the epicenter (EPI) and peri-lesioned (PERI) areas. DAPI was used to stain nuclei. Manders’ coefficient was utilized to evaluate co-staining between NeuN+ or OLIG-1+ cells and Caspase-3: the R value approximating +1 or −1 indicates linear correlation and 0 indicates absence of correlation. (E) Representative images of NeuN positive cells (green) and quantification of neurons (n = 3 mice/group) in ventral horn (VH NeuN+ cells) 30 days after SCI: DHA significantly (p < 0.05; Student’s t-test) counteracted neuronal loss. (F) Brightness values of Caspase-3 expression at the epicenter and perilesioned areas in ventral horns of spinal cord from naïve, SAL- and DHA-treated mice (n = 3 mice/group). In comparison with naïve animals, SCI induced a higher expression of Caspase-3 at the epicenter, significantly reduced by DHA treatment. After SCI Caspase-3 expression was less enhanced in the perilesioned area of SAL-treated animals, while a higher expression was present in DHA-treated mice. (G) Brightness values of OLIG-1 expression was significantly enhanced after SCI in ventral horns of spinal cord from both saline- and DHA-treated mice, mainly at the epicenter but also in perilesioned area, in comparison with naïve (n = 3 mice/group). °°p < 0.001, °°°p < 0.0001 vs naïve; **p < 0.001 vs SAL. Tukey/Kramer).