FIGURE 4.
Effect of PrPC and mGluR5 absence and of Ln-γ1 treatment on AβO toxicity hallmarks. A and B, the effect of pretreatment of neuronal cultures with Ln-γ1 or Ln-SCR (scr) on AβO-induced cell death. Cell death was registered for cultures prepared from at least five embryos for each condition (A). Representative images show live (green) and dead (red) cells (B). Scale bar = 100 μm. C and D, AβO binding to neurites lacking PrPC, mGluR5, or both proteins. Representative images showing immunostaining of AβOs (red) and tubulin (green) (C). Levels of bound AβOs were normalized to neurite area (calculated by tubulin immunostaining) from eight random fields of view of neuronal cultures prepared from at least five embryos for each genotype (D). Scale bar = 20 μm. E and F, AβO-induced increase in intracellular Ca2+ in neuron lacking PrPC (red), mGluR5 (blue), or both proteins (purple). E, calcium response (relative to baseline Fluo-4 fluorescence, F1/F0) kinetics in the neurons treated with 500 nm AβOs. F, quantification of the peak signal from E. Data were collected from at least 20 cells in at least 3 independent experiments for each genotype. Statistical comparison was done with one-way analysis of variance followed by Tukey post hoc test. Error bars indicate mean ± S.E. For a and b, p < 0.05; for a and c, p < 0.001.