Fig. 5

Wild-type cells can rescue pineal progenitor specification cell non-autonomously in otxH morphants. (A,B) Representative fluorescent images of wild-type zebrafish embryos containing cells grafted from an otxH embryo (green) and stained by in situ hybridisation for otx5 (red) at 24 hpf (lateral views, anterior is leftwards). Inset in B shows a dorsal view of the pineal region in more detail. otx5 is expressed in otxH cells that are adjacent to wild-type cells (arrow). Two independent experiments were performed grafting 13 and 19 embryos, respectively. (C) Bright-field image of an otxH embryo containing grafted wild-type cells and stained by in situ hybridisation for otx5 (red) at 24 hpf (lateral view, anterior is leftwards). (C′) Fluorescent images of the pineal region of the embryo shown in C in more detail; grafted wild-type cells are green. There is non-cell-autonomous rescue of otx5 expression (arrow). Inset in C′ shows a magnified dorsal view of the otx5-positive pineal organ in C′. Two independent experiments were performed grafting 17 and 14 embryos: 15/17 and 11/14 showed cell-non-autonomous pineal rescue. Scale bars: 50 µm in A,C′; 50 µm in insets (bar in the inset in C′ applies to the insets in both B and C′).