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. Author manuscript; available in PMC: 2013 Oct 22.
Published in final edited form as: Gastroenterology. 2008 Nov 6;136(3):902–911. doi: 10.1053/j.gastro.2008.11.012

Figure 4. elys mRNA is expressed maternally and its expression pattern becomes more restricted by 72hpf.

Figure 4

(A) Schematic diagram illustrating the positions of primers used to analyze elys mRNA expression by allele-specific (AS) RT-PCR. Asterisk indicates the position of the C→T mutation in exon 29 that provides the basis for AS primer annealing. (B) Wildtype elys is expressed continuously in wildtype embryos as expected. Maternal expression of wildtype elys mRNA in flo embryos is comparable to that of wildtype embryos until 8hpf; thereafter maternal elys expression progressively diminishes and is barely detectable at 24hpf. (C) Zygotic expression of mutant elys RNA is induced in flo from 12hpf. M=DNA markers, DW=distilled water. (D, E) WISH was used to determine the elys mRNA expression pattern from 24–96hpf. (D) elys mRNA is broadly expressed over the first 52hpf. (D, left panel) elys expression in the retina (arrow), pharyngeal region (bracket), midbrain and hindbrain (arrowhead) at 24hpf. (D, second panel) elys expression in the forebrain (arrowhead), midbrain-hindbrain boundary (arrow), liver (bracket) at 48hpf. (D, third panel) elys expression in the fin buds (arrowhead) and intestine (bracket) at 52hpf. (D, fourth panel) elys expression in the retina (arrow), pharyngeal region (arrowhead) and liver (bracket). (E) The elys mRNA expression pattern becomes more restricted at 72–96hpf. (E, first panel) elys expression in the pharyngeal region (arrowhead), liver and anterior intestine (bracket), MHB (arrow) and retina at 72hpf. (E, second panel) elys expression in the liver and intestinal bulb (bracket) and pharyngeal region (arrowhead) at 96hpf. (E, third panel) Transverse section of a WISH embryo counterstained with nuclear fast red showing elys mRNA in the cytoplasm of intestinal epithelial cells, liver and pancreas at 96hpf. (E, fourth panel) Barely detectable expression of elys in comparable regions of a flo embryo at 96hpf. Scale bar: D, E = 500µm (except E, third panel).