Fig. 3

Gas2l3 is essential for brain morphogenesis and development. (A) The zebrafish gas2l3 gene consists of 9 exons interspersed with 8 introns. Black square indicates target sequence of gas2l3 splicing-block MO. Gray arrow indicates transcript direction. Black arrows indicate forward and reverse primers designed to test MO efficiency (see Fig. S2 for more information). (B) One-cell stage embryos were injected with 2 pmol gas2l3 MO or standard control MO. Representative images of MO-injected embryos (mild or severe morphants), alongside an uninjected embryo (normal), were taken at 24 hpf. Panels ii, iv, and vi are magnified images of panels i, iii, and v respectively. Matching dorsal views are depicted in panels vii, viii, and ix. FB, forebrain; MB, midbrain; HB, hindbrain. Black arrows point toward abnormal forebrain protrusions. (C) Embryos were injected with gas2l3 MO (1 or 2 pmol) or control MO (2 pmol) in combination with mock, gas2l3 RNA, EGFP-gas2l3 RNA, and EGFP RNA (100 pg). Injected embryos were categorized as normal or abnormal according to their brain morphology at 24 hpf. Values are shown as mean normal or abnormal percentages ± SEM of abnormal. We used the ANOVA Bonferroni test in order to determine the statistical significance of the results; Means with distinct letters on top of the chart are significantly different (p<0.001, F=25.846, df=5, 31). Following microinjection, dead, mechanically damaged embryos or embryos with unspecified morphology were excluded from the analysis.