FIGURE 4.

zGas8 is important for early development and proper Hh signaling. A, zebrafish embryos injected with Gas8 MO displayed pericardial edema (arrow), hydrocephaly (arrowhead) and misshaped somites. These phenotypes can be rescued by the co-injection of a MO-resistant zGas8 mRNA and by dnPKA mRNA. Gas8 control MO (Gas8 CT MO) injected embryos were indistinguishable from the non-injected wildtype embryos (see also Fig. 5). 72 hpf embryos are shown laterally. B–D, whole mount in situ hybridization of Nkx2.2 (B), Pax2 (C), and Shh (D), showing attenuated Nkx2.2 expression in the floor plate (fp) and hind brain (hb), and Pax2 in the otic vesicle (ov) of 24 hpf embryos injected with Gas8 MO, but not with its CT MO. Normal expression levels achieved by co-injection of zGas8 or dnPKA mRNAs. Shh expression levels remained similar under all conditions. E–H, stacked bar graph analysis of the percentage of embryos with normal or abnormal phenotype (E), and the percentage of embryos that displayed attenuation or no change in the indicated transcript levels (F–H) following the different injections. Stacked bar graphs summarize at least 4 separate injections of Gas8 MO and their controls. Two of these experiments included zGas8 mRNA and the other two included dnPKA mRNA for rescue. Numbers of injected embryos are indicated in the text.