Spatiotemporal expression of dnaaf5 during zebrafish embryogenesis. (A) Schematic representation of dnaaf5 mRNA showing the position of primers used for whole-mount in situ hybridisation (WISH) probe synthesis and RT-PCR analysis. (B) Temporal expression of dnaaf5 mRNA during zebrafish development. Zebrafish ef1α is used as a loading control. (C-N) Whole-mount in situ hybridisation showing dnaaf5 transcript localisation from the 1-cell to 96 hours post-fertilisation (hpf). (C–E) During early development (1-cell to 6 hpf), dnaaf5 is ubiquitously expressed, with strong maternal deposition of the transcript and early zygotic expression. (F-N) Later, it is expressed in specific organs. (F,G) dnaaf5 is expressed in dorsal forerunner cells (DFC) and Kupffer’s vesicle (KV) at 9 hpf and 12 hpf respectively. (H-J) 18 hpf onwards it is expressed in eye, hindbrain (HB), pronephros (P) and floorplate (FP), tail bud (TB) and otic vesicle (OV). (I-N) dnaaf5 is expressed in the telencephalon (TEL), diencephalon (DI) and midbrain (MB) after 24 hpf. (K-N) Expression is more intense in the brain than in other organs by 48 hpf. (L, N) After 72 hpf, it is expressed in the liver (L). (M) Dorsal view of 72 hpf embryo head, clarifying dnaaf5 expression throughout the brain (B), retina (R) of the eye and olfactory pit (OP). (O,P) WISH with the sense dnaaf5 probe in 1 cell and 30 hpf zebrafish embryos shows no detectable signal. Scale bars, 150 µm.
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