Identification and Characterization of Highly Fluorescent Pigment Cells in Embryos of the Arabian Killifish ( Aphanius Dispar )

The Arabian killifish, Aphanius dispar, is a small tropical teleost fish living in wide range of habitats in sea water and fresh water in the Middle East. Here, we report extraordinary fluorescent pigment cells in the Arabian killifish embryo. These cells appear brown in transmitted light, yellowish white in reflected light, and as strong fluorescence in GFP and RFP filters. TEM and confocal microscopy analyses show the fluorescence emanates from leucosome-like pigment organelles. The cells express the gene encoding GTP cyclohydrolase ( gch ), a marker for leucophores and xanthophore. Gene knockdown and knockout of gch using morpholino or CRISPR-Cas9 induced loss of fluorescence in these embryos, indicating a crucial role of the enzyme and the associated pterine biosynthesis pathway in the generation of the fluorescence. We concluded that these cells are a highly fluorescent subtype of leucophores and have named them as fluoroleucophores.