Fig. 2

zC2orf62 is expressed in ciliated structures during embryonic development and highly expressed in adult testis.

(A) zC2orf62 mRNA levels measured by RT-qPCR at various developmental stages. zC2orf62 is highly expressed in adult testis and during almost all embryonic development. It is down-regulated at shield stage (6 hpf) and re-expressed at tail bud (12 hpf) when Kupffer′s vesicle forms. (B) Whole mount in situ hybridization of zC2orf62 mRNA at 24 hpf and 48 hpf. At 24 hpf, zC2orf62 mRNA is detected at the end of neural tube formation on the tail extremity (arrowhead), in pronephric ducts (P.D.) and in the brain (arrows). At 48 hpf, zC2orf62 is expressed in forebrain (B) and olfactory pits (O.P.) (arrows). Scale bars, 100 μm. (C) A construct containing zC2orf62 52-UTR, 32-UTR and potential regulatory sequences in which zC2orf62 coding sequence was replaced by EGFP was generated and inserted randomly inside the zebrafish genome. Resulting transgenic EGFP-zC2orf62 reporter fish were observed at 12 hpf using a fluorescent stereomicroscope and at 28, 48 and 96 hpf using a confocal microscope. EGFP is expressed at 12 hpf in Kupffer′s vesicle, and at 28 hpf in olfactory placode, eyes, brain, neural tube (N.T), and pronephric ducts (P.D). At 48 hpf, EGFP is expressed in ciliated cells of the olfactory organ (positive for μ-acetylated tubulin (red)). At 96 hpf, EGFP is also expressed in neuromast cells (N), olfactory sensory neurons (OSN) and in the ears (E and lower enlarged panel). Scale bars, 25 μm. See also Figs S1, S2.