Fig. 4

Fig. 4

Embryonic inhibition of col8a1a function or lysyl oxidases is sufficient to generate vertebral defects. (A) Timeline of vertebral column development in zebrafish. In one experiment, the injection of MO2-col8a1a at the 1-cell stage, is effective in blocking the splicing of col8a1a until~3–5dpf (purple gradient bar) (C–C′′). In the second experiment the lysyl oxidases inhibitor, βAPN, is added at 4hpf and washed out at 1dpf (orange bar) (D–D3). Both treatments completed well before the onset of centra formation started in the anterior regions at 5dpf. Representative brightfield images of 1dpf (B, C, and D) and 60dpf (B′, C′, D′) and microCT images (45dpf) (B′′, C′′, D′′) of WT (B–B′′), MO2-col8a1a injected (C–C′′), and βAPN pulsed adult zebrafish (D–D′′). (D) Categorical frequency of VM for 2 and 4 ng MO2-col8a1a morpholino injected adults and uninjected WT siblings and 2, 4, and 10 mM βAPN treated adults and DMSO treated sibling controls for both thoracic (T) and caudal (C) regions. Scale bars for (B, C, and D)=4 mm, (B′, C′, D′)=5 mm, and (B′′, C′′, D′′)=1 mm.