Journal: Scientific Reports
Article Title: EAAC1 gene deletion reduces adult hippocampal neurogenesis after transient cerebral ischemia
Figure Lengend Snippet: Genetic deletion of EAAC1 reduces survival of newly generated neurons after ischemia. ( A ) The schematic diagram shows the experimental procedure used in this study. BrdU was intraperitoneally injected twice per day for 4 consecutive days from day 3 after ischemia. Mice were killed on day 30. ( B ) Photomicrographs show the distribution of BrdU-labeled cells in the SGZ/GCL of the hippocampus of young and aged mice (WT or EAAC1 −/− ) after ischemia. Newborn cells were labeled with BrdU, and their survival was assessed 30 days after ischemia. Scale bar = 100 μm. ( C ) Graph shows the number of BrdU-labeled cells in the SGZ/GCL of young and aged mice (WT or EAAC1 −/− ) at 30 days after ischemia. Data are expressed by the mean + SEM; n = 3–7 from each group, * p < 0.05 versus WT mice; # p < 0.05 versus young mice. (D,E) Representative immunofluorescence images show the phenotype of cells that have survived in the SGZ/GCL. Scale bar = 20 μm. ( F , G ) Bar graph represents the number of BrdU-labeled cells that were colocalized with NeuN (F) or GFAP (G) in the SGZ/GCL from young and aged mice (WT or EAAC1 −/− ) at 30 days after ischemia. Data are expressed as the mean + SEM; n = 3–7 from each group, * p < 0.05 versus WT mice; # p < 0.05 versus young mice.
Article Snippet: We used the following antibodies as primary antibodies; guinea pig polyclonal anti-DCX (diluted 1:1k, Millipore), mouse monoclonal anti-beta 3 tubulin (TUJ1, diluted 1:500, Abcam), mouse monoclonal anti-microtubule-associated protein 2 (MAP2, for recognizing a neuron-specific cytoskeletal protein, diluted 1:200, Millipore), and rabbit polyclonal anti-excitatory amino acid transporter 3 ( EAAT3 , also known as EAAC1 , diluted 1:200, Almone labs, Jerusalem, Israel).
Techniques: Generated, Injection, Labeling, Immunofluorescence