PKCδ promotes fertilization of mouse embryos in early development via the Cdc25B signaling pathway
2019
Protein kinase C type δ (PKCδ) is involved in B-cell signaling and the regulation of growth, apoptosis and differentiation of a variety of cell types. Cell division cycle 25 (Cdc25) is a key mediator of cell cycle progression that activates cyclin-dependent kinase complexes that drive the cell cycle and participates in the regulation of DNA damage checkpoints. Cdc25B is a member of the Cdc25 family of phosphatases. The present study investigated the role and mechanism of PKCδ in regulating the fertilization of mouse embryos in early development. The expression and subcellular localization of PKCδ and Cdc25B were detected using reverse transcription-quantitative polymerase chain reaction, western blotting and immunofluorescence in one-cell stage mouse embryos. Specific small interfering RNAs targeting PKCδ were used to knockdown the expression of PKCδ. Subsequently, Scansite software was used to predict the target of phosphorylated Cdc25B. Western blotting was used to measure the effects of phosphorylation and dephosphorylation in one-cell stage mouse embryos at different cell cycle phases. PKCδ was expressed during M phase and served a positive role in one-cell stage mouse embryos. Immunofluorescence data revealed that PKCδ and Cdc25B were expressed during G1, S, G2 and M phases of the cell cycle. Furthermore, phosphorylated levels of Cdc25B-Ser96 were observed during G2 and M phases. Microinjection with mimics of phosphorylated Cdc25B-Ser96 mRNA promoted the development of one-cell stage mouse embryos. When PKCδ was suppressed, microinjection with mimics of phosphorylated Cdc25B-Ser96 mRNA reversed the inhibition of PKCδ. To conclude, PKCδ serves a positive role in the first cell cycle of mouse embryos by phosphorylating Cdc25B-Ser96, and provides novel insights for the regulation of early embryonic development.
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