Reverse Genetics Screen in Zebrafish Identifies a Role of miR-142a-3p in Vascular Development and Integrity
A - Schematic of the experimental approach. MiRNA duplexes were ectopically overexpressed in Tg(fli1:EGFP, gata1a: dsRed) zebrafish embryos through microinjection at 1–2 cell stage. The injected embryos were scored at 2 dpf under microscope for visual observation of phenotype under bright field, GFP and RFP filters. Representative images of miRNA-injected Tg(fli1:EGFP, gata1a: dsRed) zebrafish embryos are displayed. B,C,D - Non-injected control (NIC) zebrafish embryos with normal vascular development. E,F,G - Zebrafish embryos injected with miRNA annealing buffer display normal vascular development. H,I,J - Zebrafish embryos injected with miR-144 display reduced or absence of blood in intersegmental vessels. K,L,M - Zebrafish embryos injected with miR-1 display accumulation of blood cells in LDA/YSL. N,O,P - Zebrafish embryos injected with miR-142a-3p display pooling of blood cells in head/trunk region. Q,R,S and T,U,V - Zebrafish embryos injected with miR-181a and miR-181b respectively display no visible phenotype. Zebrafish embryos injected with miR-221, miR-222 and miR-451 display no observable phenotype (figure not shown). Arrowheads indicate the site of vascular defects. The embryos were imaged at 2.5× magnification.