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Fig. 1 Gpr161 is an evolutionarily conserved ciliary GPCR that is essential during embryonic development. (A) gpr161a and gpr161b transcript levels at different stages of development analysed in whole-embryo lysates in triplicate using RT-qPCR. (B) Wild-type embryos were injected with gpr161a-myc or gpr161b-myc mRNA at the one-cell stage and fixed at 9?hpf before immunostaining for acetylated tubulin (AcTub; purple), a marker for the ciliary axoneme, and Myc (green). Scale bar: 5?µm. (C) Lateral views of wild-type and MZgpr161b?/?; gpr161a?/? embryos at 24?hpf. Insets show ventral views of the developing eyes; arrowheads indicate the otic vesicle. Scale bars: 100?µm. (D) Wild-type and MZgpr161b?/?; gpr161a?/? embryos at 72?hpf. Arrowheads indicate otic vesicle. Scale bars: 200?µm. Insets show a dorsal view of the head. (E) Ventrolateral views of the craniofacial region of wild-type and MZgpr161b?/?; gpr161a?/? embryos at 72?hpf taken by scanning electron microscopy; arrowheads indicate the olfactory pit. Scale bar: 100?µm. (F) Transverse semi-thin sections of the eye in wild-type and MZgpr161b?/?; gpr161a?/? embryos fixed at 72?hpf (Richardson staining). Scale bar: 50?µm.