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Journal: The European journal of neuroscience
Article Title: Expression pattern of miR-146a, an inflammation-associated microRNA, in experimental and human temporal lobe epilepsy.
doi: 10.1111/j.1460-9568.2010.07122.x
Figure Lengend Snippet: Fig. 2. In situ hybridization analysis of miR-146a expression in hippocampal tissue of control rats and after induction of SE. (A, C, E, G) Control hippocampus showing neuronal miR-146a expression in the different hippocampal subfields, including pyramidal neurons of CA3 (C; insert in C) and CA1 (E; insert in E) regions, granule cells of the dentate gyrus (DG; G) and hilar neurons (G; insert in G: high-magnification of hilar neurons). gcl, granule cell layer. (B, D, F, H) Hippocampus 1 week post-SE showing increased miR-146a expression within the different hippocampal regions, including CA3 (D; insert in D: high-magnification of CA3), CA1 (F; insert in F: high-magnification of CA1) and DG (H). miR-146a expression was observed in both neuronal and glial cells; arrows in D, F (and insert in D) indicate positive pyramidal neurons of CA3 and CA1; positive glial cells (arrowheads in D, F and insert in D) were particularly abundant in regions of prominent gliosis (CA1, CA3 and the inner molecular layer, iml, of the DG). Insert in (H) shows a high-magnification of positive glial cells in the DG (iml). (I and J) In situ hybridization analysis of miR-146a expression in the hilar region of the hippocampus 1 week (I) and 3–4 months post-SE (J) showing increased expression in glial cells (arrows; insert in I). Sections are counterstained with haematoxylin. Inserts in (J): in situ hybridization and immunohistochemistry analysis showing in (a) absence of miR-146a (red) expression in lectin-positive microglial cells (blue) and in (b) colocalization with the astroglial marker glial fibrillary acidic protein (GFAP; purple) in astrocytes. Scale bars: 1250 lm (A and B); 70 lm (C–J); 35 lm (inserts in C, D, F and G); 20 lm (insert in E); 17 lm (inserts in H and I); 11 lm (insert in J). LT, long-term, 3–4 months after SE.
Article Snippet: For the double-staining, combining immunocytochemistry with in situ hybridization, sections were first processed for immunocytochemistry as previously described (Aronica et al., 2001a, 2003) with glial fibrillary acidic protein (GFAP; polyclonal rabbit; DAKO, Glostrup, Denmark; 1 : 4000), neuronal nuclear protein (NeuN; mouse clone MAB377; Chemicon, Temecula, CA, USA; 1 : 2000), HLA-DR [anti-human leukocyte antigen (HLA)-DP, DQ, DR (mouse clone CR3 ⁄ 43); DAKO, Glostrup, Denmark; 1 : 400], CFH (polyclonal goat; Quidel, San Diego, CA, USA; 1 : 100) or the
Techniques: In Situ Hybridization, Expressing, Control, Immunohistochemistry, Marker