Fig. 5

Fig. 5 Whole-mount immunostaining of the otoliths of wild type and MO-injected embryos. (A, D, G, J) Live embryos; all other panels show fixed, immunostained embryos. (A–C) Wild-type embryos at 27 hpf; (B) zOMP-1 is detected in both otoliths and in the ventro-lateral epithelium posterior to the anterior macula (arrowheads), but zOtolin-1 is not observed yet (C). (D–O) Embryos at 72 hpf. In control MO-injected embryos, anti-OMP-1 (E) and anti-Otolin-1 (F) antibodies recognize both otoliths. (G–I) zomp-1 MO-injected embryos; zOMP-1 is effectively knocked down (H); the anti-Otolin-1 antibody fails to detect zOtolin-1 in the small otoliths (I). (J–L) zotolin-1 MO-injected embryos; zOtolin-1 is not detected (K) while the anti-OMP-1 antibody reacts strongly with the fused otoliths (L). (M–O) Pre-immune serum shows non-specific staining (arrowheads) of the sensory hair cells of the anterior macula (M), posterior macula (N) and anterior and lateral cristae (O). In panels (A–G), the posterior otoliths were inlayed from a deeper focal plane onto the main pictures, which are focused on the anterior otoliths. Dotted lines outline otoliths in panels (H), (I) and (K) and the posterior macula in (N). Through the panels, anterior is to the left, dorsal to the top. Scale bar in (A) and (O) indicates 25 μm in (A–N) and (O), respectively.