Developmental expression and possible functional roles of mouse Nlrp4e in preimplantation embryos

Increasing evidence suggests that some Nlrp genes are crucial for oogenesis, folliculogenesis, and early embryonic development. Nlrp4e is one of seven copies of Nlrp4, which plays a putative role in the reproduction system in mice. Gene duplication is regarded as an important driving force behind the evolution of novel genes with new or altered functions. We investigated the role of Nlrp4e in oocyte and preimplantation embryos by determining its expression profile using quantitative real-time polymerase chain reaction. Nlrp4e mRNA accumulated during oogenesis. Moreover, Nlrp4e transcripts were upregulated during the two-cell stage and then declined sharply and became almost undetectable, which represents a crucial time for major embryonic genome activation in the mouse. Knockdown of Nlrp4e in fertilized eggs using RNA interference resulted in arrested development between the two- and eight-cell stages in a dose-dependent manner. However, targeted inhibition of Nlrp4e in germinal-vesicle-stage oocytes had no phenotypic effects on oocyte maturation. The above experiments were also carried out in parthenogenetic embryos to determine the effects of Nlrp4e in embryos without a paternal genome. The results of this study indicate that Nlrp4e, a maternal-zygotic-effect gene, may not be involved in oocyte maturation but may play a critical role in early embryogenesis.