hdac3 is specifically required for liver formation in zebrafish. (a) hdac3 was knocked down by antisense morpholino (12 ng/embryo) in Tg(lfabp:RFP; elaA:EGFP) and liver formation was observed from 3 dpf to 5 dpf. Control (Con.) embryos were injected with 5 bp mismatch antisense morpholino. Blood vessels formation was monitored in Tg(fli-1:EGFP) at 3 dpf. Liver (thick white arrow) was present at 3 dpf, but its size is much smaller (A vs. B). In addition, it failed to grow from 3 dpf to 5 dpf or reduced growth was observed compared to control embryos (E–H). ISV (thin white arrow) was disrupted. All the images are lateral view, anterior to the left. Scale bar represents 100 μm for all panels except panels C and D which is 50 μm. (b) Liver formation in hdac3 morphants monitored by WISH with liver marker Ceruloplasmin (Cp) at 32 hpf (A, B), 2 dpf (C, D), 3 dpf (E, F) and 5 dpf (G, H). In hdac3 morphants, Cp expression (white arrow) was absent up to 2 dpf (A, D) whereas a reduced liver was observed at 3 dpf (E, F) and 5 dpf (G, H). All images are dorsal view, anterior to the left. Arrowhead indicates Cp expression in YSL. (c) hdac3 is not involved in pancreas formation. Exocrine and endocrine pancreas were monitored by GFP in Tg(lfabp:RFP; elaA:EGFP) and WISH with insulin, respectively. Control embryos were injected with same amount of 5 bp mismatch morpholino. Both exocrine pancreas (white arrowhead, A–D) and endocrine pancreas (black arrow, E–J) were not affected in hdac3 morphants, whereas liver (red fluorescence, white arrow) was affected (A vs. B). All the images are dorsal view, anterior to left. Scale bar is 50 μm.
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