Fig. 6.

Toxicity of LCBs to cultured neurons. (A–L). Dihydrosphingosine (dhSph) treatment of cortical neurons. Cortical neurons from E18 embryos were untreated (A–D), or treated with 1 µM (E–H) or 15 µM (I–L) dhSph. (M–P) Deoxysphinganine (DeoSa) treatment of cortical neurons. Neurons were treated with 0.15 µM DeoSa. An area is enlarged in the Inset in P to show axon fragmentation at its early stage (arrowheads). The rationale for using these dhSph and deoSa concentrations was explained in detail in the Results. Neurons, after fixation, were incubated with an antibody against neuron-specific class III β-tubulin (A, E, I, and M; Tuj1), and then subjected to TUNEL assay (B, F, J, and N) and counterstained with DAPI (C, G, K, and O). Merged images were shown (D, H, L, and P). (Scale bar, 20 µm.)