Mature neurons were identified with Anti-NeuN antibody in control and AβO microinjected rats sacrificed on days 15 and 30 after stereotaxic surgery (n = 4, per group and time). For degenerated neurons, the FJB staining was evaluated in control and AβO groups at 18 h, 72 h, and 7 days post-surgery (n = 4, per group and time).
Quantification of NeuN and FJB positive cells was realized with 8 µm-thick sagittal sections obtained from injury (2.5–2.7 mm lateral from bregma), one out every 10 sections was evaluated. Cell count was performed on the Leica Application Suite software version 4.0 (Leica Microsystems, Wetzlar, Germany) on a 265 × 365 µm frame at 40× magnification, analyzing 3 adjacent fields on CA1 region and the same for dentate gyrus. The experimental groups were blinded in during the count and we considered positive cells with whole cell bodies within the counting frame. Average of positive cells per mm2 were reported [80 (link),81 (link)].
Quantification of NeuN and FJB positive cells was realized with 8 µm-thick sagittal sections obtained from injury (2.5–2.7 mm lateral from bregma), one out every 10 sections was evaluated. Cell count was performed on the Leica Application Suite software version 4.0 (Leica Microsystems, Wetzlar, Germany) on a 265 × 365 µm frame at 40× magnification, analyzing 3 adjacent fields on CA1 region and the same for dentate gyrus. The experimental groups were blinded in during the count and we considered positive cells with whole cell bodies within the counting frame. Average of positive cells per mm2 were reported [80 (link),81 (link)].
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