Fig. 5

Fig. 5

A Cartoon schematic of zebrafish larvae showing the approximate location of sparsely labeled clusters of neurons. B Neurons in the zebrafish larvae spinal cord co-expressing NTR-BFP, EGFP, and RGEDI at 0, 24, and 48 h after mounting for automated imaging in MTZ. White arrows indicate neurons co-expressing NTR-BFP and EGFP. Scale = 50 μm. C Quantification of GEDI ratio in neurons co-expressing NTR-BFP and RGEDI-P2a-EGFP exposed to MTZ (n = 7) or DMSO (n = 5) showing no increase in GEDI signal (ANOVA Sidak’s, ns not significant). D Neurons in the zebrafish larvae spinal cord co-expressing NTR-BFP, mApple, and GC150 at 0, 24, and 48 h after mounting for automated imaging in DMSO. White arrows indicate neurons with the fluorescence of BFP and mApple, asterisks indicate autofluorescence from pigment in larvae skin, scale = 20 μm. E Quantification of average GEDI ratio of neurons from zebrafish larvae incubated in DMSO (n = 7) or 10 μM MTZ (n = 17) over time showing an increase in GEDI ratio in neurons indicating neuronal death after 24 h in MTZ (ANOVA Tukey’s ****p < 0.0001, ***p < 0.0005). F Neurons in zebrafish larvae spinal cord co-expressing NTR-BFP, mApple, and GC150 at 0, 24, and 48 h after mounting for automated imaging in 10 μM MTZ. White arrows indicate neurons with the fluorescence of BFP and mApple, yellow arrows indicate neurons with the fluorescence of BFP, mApple, and GC150, indicating neuronal death. Asterisks indicate autofluorescence from pigment in larvae skin, scale = 20 μm. Error bars represent SEM. The experiment was on 12 larvae with similar results.