Figure 1

Microglia-coculture induces alterations in cellular morphology in cultured Müller cells. (A) Morphological features of cultured mouse Müller cells were visualized by immunolabelling of the actin cytoskeleton (with phalloidin, red), cytoplasm (with glutamine synthetase (GS), green), and nuclei (with DAPI, blue). Representative examples of Müller cells cultured alone (top), co-cultured with mouse microglia (middle), and cultured with microglia previously activated with lipopolysaccharide (LPS, 1 μg/ml) (bottom), are shown. Müller cells cultured alone were similar to those co-cultured with unactivated microglia in having broad lamellipodia and a symmetrical cell shape (top and middle rows) while co-cultured with activated microglia (bottom row) had elongated spindle (arrowhead) or multipolar (arrow) morphologies. Quantitative shape analyses in terms of morphological parameters of cellular area (B), perimeter (C), circularity (D) and elongation factor (E) revealed significant decreases in cellular area and increases in cellular elongation, without changes in overall cellular perimeter (n = 348 to 363 cells per group, * indicates p values < 0.05 for comparisons, one-way ANOVA).