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538979.fig.002b
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Figure 2: Hypertrophy of astroglia and a loss of dendritic spines following 3-day letrozole treatment. (a) Gender mixed hippocampal neural cells were isolated from 3-day old C57/BL6 mouse pups and fixed in 10% Formalin on DIV7. Shown is confocal images of immunocytochemically labelled hippocampal cell cultures with GFAP (astroglia, green) and β-III tubulin (neurons, red). Arrowheads outline astrocytic hypertrophy in letrozole-treated cultures after 72 hours. Scale bar, 20 μm. (b) Representative 3D reconstructions of dendritic segments from organotypic hippocampal sister slice cultures that were treated with either control or letrozole (1 μm)-containing media for either 24 or 72 hours. Scale bar, 2 μm. (c) Quantification of the spine subtype densities following treatments. There is a significant decrease in the densities for all spine subtypes following letrozole treatment compared to control. 24-hour treatment, control, n = total dendritic segment lengths of 441 μm from 10 cells in 4 cultures; letrozole, μm of dendrite from 9 cells in 4 cultures. 72-hour treatment, control, μm of dendrite from 12 cells in 4 cultures; letrozole, μm of dendrite from 11 cells in 4 cultures.