Expression of clint1 in epidermal tissues and reduced clint1 expression, inflammation and epidermal defects in hi1520 mutants. (A) Zebrafish clint1 locus showing the hi1520 insertion. (B) RT-PCR amplification of clint1, il1b and ef1a from wild-type embryos (1) and from heterozygous (3,5) and homozygous (2,4,6) clint1 mutant embryos. Arrow and asterisk indicate full-length and alternatively spliced clint1 transcripts, respectively. (C-E) Whole-mount in situ hybridization (WISH) of clint1 (purple) in wild-type (WT) embryos at 36 hpf showing epidermal expression. (C) clint1 expression within lateral epidermis (arrowhead) and fin fold (arrow). Lateral view (D) and transverse sections (E) of embryos subjected to WISH for clint1 (purple) and p63 immunolabeling (brown) showing clint1 expression in both p63-positive (arrows) and p63-negative (arrowheads) cells. (F,F′) WISH of mpo in wild-type (F) and in clint1 mutant (F′) embryos. (F) Anterior (arrow) and posterior (arrowhead, CHT) hematopoietic tissues of wild-type sibling. (F′) Dorsal ridge (arrows) and caudal fin fold (arrowhead) of clint1 mutants. (G,G′) Wild-type (G) and clint1 mutant (G′) embryos at 48 hpf showing cellular aggregates (arrowheads). (H-J′) Hematoxylin and Eosin-stained transverse sections taken anterior to cloaca of wild-type (H-J) and clint1 mutant (H′-J′) embryos at 48 hpf. Red and blue boxed regions in H,H′ are magnified in I,I′ and J,J′, respectively. Arrowheads (I′,J′) identify protruding epidermal cells. YSE, yolk sac extension. Scale bars: 200 μm.
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