Figure 5

DHA potentiates Iduna expression in neurons and astrocytes. Immunohistochemistry on days 1, 3 and 7 after MCAo. Iduna-positive cells were counted in the cortex at two bregma levels: (a) +1.2 mm (A1, A2, A3) and (b) −2.8 mm (P1, P2, P3; see diagrams on the top). DHA treatment increased Iduna expression in neurons on day 1 and in astrocytes on days 1, 3 and 7. (c) Iduna expression in the neurons and astrocytes from the ischemic cerebral cortexes (see diagram) of vehicle- and DHA-treated rats on day 1. (d) Iduna expression in the neurons and astrocytes from the contralateral cerebral cortex (see diagram on the top) from vehicle- and DHA-treated rats on day 1. Iduna expression was mostly located in the cytoplasm of the neurons and also expressed in astrocytes. (e) Immunohistochemistry shows a perinuclear Iduna expression compared with vehicle-treated animals (picture inside the white box) on days 1 and 7 after DHA treatment. Iduna expression was observed in the nucleus and projections of the neurons on day 1. Contrary to this, perinuclear localization of Iduna occurred on day 7. (f) Western blot: The quantitative data of Iduna expression from ipsi- and contralateral areas on days 1 and 3, and (g) cortical and subcortical areas (see diagram) on days 1 and 3 and cortex on days 1, 3 and 7. DHA treatment upregulated the expression of Iduna on day 1 and 3. (g) Immunohistochemical detection of Iduna. (h) Western blot of ipsilateral penumbra on day 1 from MCAo treated with saline, MCAo treated with DHA and sham-MCAo (no MCA occlusion) treated with DHA groups. Data are mean±S.E.M. *, significantly different from saline (P<0.05; repeated measures analysis of variance (ANOVA) followed by Bonferroni tests). Saline n=7; DHA n=6. Values shown are means±S.D. *P<0.05, versus saline group (repeated measures ANOVA followed by Bonferroni tests). Iduna (green); NeuN (red), magnification × 40. Neurons (arrows) and astrocytes (arrowheads)