Knockdown of zebrafish Nanog increases primordial germ cells during early embryonic development

Nanog is a homeodomain transcription factor that plays a prominent role in maintaining the pluripotency and self‐renewal capacity of embryonic stem cells (ESCs) in mammals. Medaka Nanog is necessary for S‐phase transition and proliferation during embryonic development. However, whether Nanog regulates the proliferation of primordial germ cells (PGCs) during embryonic development has not yet been investigated. In this study, we identified the homologue of the mammalian Nanog gene in zebrafish (zNanog). The expression of both zNanog mRNA and protein was demonstrated in the spermatogonia (male germ stem cells) of the testis and the early oocytes of the ovary. During the embryonic development, zNanog mRNA is expressed in the cytoplasm of PGCs, and its protein is localized to the PGC nuclei. We also found that zNanog depletion using morpholinos resulted in the increases and aberrant localization of PGCs in the zebrafish embryos from the sphere stage to the 50% epiboly stage. These data indicated that zNanog inhibits the PGCs proliferation in early embryonic development of zebrafish. Using Vasa RNA probes, whole mount in situ hybridization and quantitative RT–PCR to study the functions of zNanog, we demonstrated that zNanog depletion resulted in the increases and aberrant localization of PGCs in the embryos from the sphere stage to the 50% epiboly stage. These data indicated that zNanog inhibits the PGC proliferation in early embryonic development.