Dnd is required for primordial germ cell specification in Oryzias celebensis

Dead end (dnd) is a germ plasm component that plays an essential role for primordial germ cell (PGC) development in vertebrates. Previously, we have found that dnd is the first fish PGC specifier in medaka. Here, we present an additional evidence that dnd is the determinant for PGC specification in Oryzias celebensis. In adult tissues, the O. celebensis dnd (Ocdnd) RNA shows germ cells specific expression in gonads. In the testis, Ocdnd RNA is strongly detected in spermatogonia and meiotic cells and gradually decreases during the spermatogenesis. In the ovary, Ocdnd RNA is present throughout oogenesis. In the embryos, Ocdnd RNA is maternally provided and asymmetrically localized to prominent particles of presumptive PGCs before gastrulation stage and restricted to PGCs subsequently. In addition, Ocdnd 3' UTR can induce specific and stabilized GFP reporter expression in PGCs. Furthermore, knockdown of Ocdnd by morpholino (MO) injection abolishes the PGCs formation and this can be rescued by co-injection of medaka dnd (Oldnd) mRNA. More importantly, overexpression of Oldnd mRNA surprisingly boosts PGCs number. These results provide insights into function of dnd as a conserved specifier of PGCs in the genus Oryzias.