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Fig. 2 | Fluids and Barriers of the CNS

Fig. 2

From: Microglia activated by microbial neuraminidase contributes to ependymal cell death

Fig. 2

Viability of ependymocytes in ventricular wall explants co-cultured with NA-activated microglia. Septal and striatal explants with an intact ependymal cell layer were obtained from the lateral ventricles of adult rats. The explants were exposed to microglial cells, either resting (b) or stimulated with LPS (c) or NA (d). Some explants were exposed to NA in the absence of microglia (a). After 24 h, explants were stained with trypan blue, fixed, paraffin-embedded and sectioned. Dead ependymal cells were stained blue (arrows in a, c and d), and were easily distinguishable from alive cells, which appeared purple due to haematoxylin staining. Live and dead ependymal cells were counted, and viability was expressed as the percentage of living cells (e). In explants cultured alone and treated with NA (a) and in those co-cultured with non-stimulated microglia (b), only few dead ependymal cells could be found (arrows); ependymal cell viability was similar in both conditions (e). However, in those explants co-cultured with microglia activated either with LPS (c) or with NA (d) the ependymal layer appeared partially disrupted, with more dead cells (arrows in c and d) and some nude spaces probably due to detached cells (arrowheads in c and d). The co-culture of the explants with microglia activated with NA or with LPS provoked a similar decrease of ependymal cells viability, compared to the viability in explants only exposed to NA or cultured with non-stimulated microglia (e). Bars in histogram represent mean ± s.d. of n = 4–6 independent experiments with explants and microglial cultures obtained from different rats. Means were compared by one-way ANOVA (P = 0.007). Letters a and b on top of the bars indicate the absence (if the same letter appears) or existence (if different letters appear) of a significant difference between the compared conditions, as indicated by Mann–Whitney test (P < 0.005). Ex: explant; NA: neuraminidase added to medium; LPS: lipopolysaccharide added to medium; (-): control medium. Scale bar: 25 μm

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