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Fig. 5
TNT in pink water increases the number of apoptotic cells in zebrafish embryos at 35 hpf. (A) Bright field images (a,b) and fluorescence images of acridine orange staining (a′,b′) by confocal microscopy show apoptosis in live whole embryos following treatment from 5–35 hpf with pink water containing TNT at 0 (a,a′) and 13.5 µg/mL (b,b′). Four tiled confocal images were combined to show the whole embryo in each case. Scale bar 500 µm. TNT treatment causes defective development of blood island and abnormal blood circulation (b, arrow) and more apoptosis in actively developing tissues of the tail (b′, arrowhead) compared with control (a′, arrowhead); (B) Magnified view of the tail regions of the embryos in (A) showing apoptotic cells (green dots); (a) is from A (a′) and (b) is from A (b′) respectfully. Scale bar 150 µm; (C) Number of acridine orange-positive apoptotic cells (AO + cells) in the boxed area in actively developing tail tissue (B, arrowhead) was counted at different concentrations of TNT using an Axioimager II fluorescence microscope. Cell death in TNT-treated embryos increased in a dose-dependent manner. n = 10 for each treatment, * p < 0.001.