SP600125 reduced the dopamine neurons loss in scamp5a KO zebrafish. A, B larvae injected with DMSO or 5 μM SP600125 at the one-cell stage. Brains extracts from 5 dpf larvae as indicated were harvested for Western blot analysis. A Lysates were probed with indicated antibodies. The primary antibody was anti-p-JNK, anti-t-JNK. GAPDH was used as a loading control. n = 20 larvae per group for proteins extracts. B Quantification of relative expression levels of apoptosis-related factors compared to those in control group. C, D Representative images (C) and relative analysis (D) of the th1-positive dopamine neurons level in 72 hpf larvae. Zebrafish treated with DMSO: WT (n = 27) and scamp5a−/− larvae (n = 31) and Zebrafish treated with SP600125: WT (n = 30) and scamp5a−/− larvae (n = 34). Dopamine neurons are indicated by whole-mount in situ hybridization (WISH) staining of th1 mRNA. Scale bars: 100 μm. Inhibition of JNK rescued dopamine neurons loss in scamp5a KO zebrafish brain. E RT-qPCR analysis of the transcript level of th1 in 5 dpf larvae brains treated with DMSO or SP600125. The relative transcript level of th1 in scamp5a−/− larvae was compared to that in WT treated with DMSO. Housekeeping gene actb was used as internal control for RT-qPCR assay. n = 30 larvae per group for mRNA extracts. The data in panel B and D are presented as mean ± SEM. n ≥ 3 independent experiments. Significance levels are denoted as follows: ***P < 0.001. ns not significant
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