FIGURE 2.

Isolation of a functional mct8 promoter and generation of a Tg(mct8:EGFP) stable transgenic fish. A, bioinformatic analysis of putative 2000-bp orthologous mct8 promoters from zebrafish, mouse, rat, and human. Oval color-coded marks drawn above or below the black and gray bars indicate the sense or antisense direction of the transcription-factor binding sites, respectively. Purple represents binding sites for the purine-rich single-stranded DNA-binding protein α (PURA). Pink represents binding sites for Huntington disease gene regulatory region binding proteins (HDBP). Green represents putative binding sites for GATA. B, schematic illustration of the pT2-mct8:EGFP DNA construct that was used to generate the Tg(mct8:EGFP) transgenic line. C-I, Tg(mct8:EGFP) transgenic larvae. C, lateral view of a 24-hpf embryo. EGFP expression driven by the mct8 promoter is observed in the CNS, including the forebrain (FB), midbrain (MB), hindbrain (HB), and along the spinal cord (SC). EGFP is also expressed in the eyes and in the notochord (No). D, dorsal view of the head of a 4-dpf larva. E, lateral view of a 3-dpf larva. EGFP is expressed in the CNS, eyes, otic vesicle (OV), and heart (He). F, lateral view of the spinal cord (SC) and the notochord (No) of a 48-hpf embryo. G, lateral view of the otic vesicle of a 4-dpf larva. White arrows point to epithelial cells located above the otic vesicle. H, dorsal view of a 4-dpf larva. EGFP expression is observed in the posterior cerebral veins (PCeV) and in cells within the choroid plexus (CP). I, dorsal view of the olfactory bulbs (OB) of a 4-dpf larva.