Clq-like Factor, a Target of miR-430, Regulates Primordial Germ Cell Development in Early Embryos of Carassius auratus

摘要

Clq-like is a significant maternal factor of TNF/Clq super-family, and the abundant protein has been observed in both mature eggs of Carassius auratus and Carassius auratus gibelio, but its biological function in early embryo development has remained unclear. In this study, we firstly revealed a high level of maternal Clq-like transcript existence only in mature eggs of Carassius auratus, whereas no any maternal Clq-like transcript was observed in that of Carassius auratus gibelio. During embryonic development the Clq-iike zygotic expression begins around cardiopalmus stage in embryos of both Carassius auratus and Carassius auratus gibelio. Then, we examined the biological role of Clq-like by morpholino-mediated knockdown in early embryo development. Knockdown of CaOCIq resulted in a significant reduction of primordial germ cells (PGCs) in Carassius auratus, as shown by whole mount in situ hybridization with voso-specific RNA probe, fluorescence im-munostaining of vasa protein, and GFP imaging of the GFP-nanosl-3'UTR mRNA reporter. In vitro and in vivo evidence indicated that a microRNA, miR-430 could repress the Clq-like expression and PGC development. These data suggest that Clq-like should be a direct target of miR-430 and play an essential role in PGC development of Carassius auratus.