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ids mutant fish-derived primary neurons show short axons and impaired lysosomal acidification. A Lysotracker staining (red) on wild-type and ids mutant fish-derived primary cell cultures at 2 DIV. Concanavalin (green) and Hoechst staining (blue) are reported to visualize cell membranes and nuclei, respectively. Magnifications of the Lysotracker-stained cells with reduced signal in mutant fish-derived cells are shown. On the right, bar graphs represent the area and integrated density quantifications normalized to cell number. In both cases, a significant decrease is measured in mutant fish-derived primary neurons when compared to controls (n = 4 independent experiments). B On the left, a scheme of the exploited microfluidic device is depicted. It is constituted by four wells and two main compartments divided by a microgroove barrier. Cells are seeded in one chamber and axons extend towards the other one. On the right, representative confocal images of wild-type and ids mutant fish-derived primary neurons seeded in the microfluidic device (2 DIV). Cultures were stained with concanavalin to easily trace processes. Below, magnifications of crossing axons are shown. The graph on the right resumes the results obtained by measuring crossing axons lengths (n = 3 independent experiments). ids mutant fish-derived neurons presented consistently shorter axons when compared to controls at 2 DIV. Scale bar: 50 μm. Data are expressed as the mean ± SD (*p < 0.05; t-test).
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