NEK2 supports porcine embryonic development by modulating the AKT signaling pathway

Abstract

Aims: Serine/threonine kinase NIMA-related kinase 2 (NEK2) plays a crucial role in regulating the cell cycle and DNA damage response. This study explored the mechanisms by which NEK2 inhibition affects porcine embryonic development. Materials and methods: To explore the role of NEK2 in porcine embryonic development, we used the NEK2 inhibitor JH295 and the AKT activator SC79. Various staining methods, including EdU, EU, OPP, TUNEL assay, real-time PCR, immunocytochemistry, and Western blotting, were used to identify the effects of NEK2 inhibition on developmental competence, DNA damage, and the related mechanisms in porcine embryos. Key findings: NEK2 inhibition significantly reduced the cleavage rate and blastocyst formation rate. Abnormal development was associated with decreased expression of genes related to zygotic genome activation and significantly reduced the levels of EdU, EU, and OPP. Notably, NEK2 inhibition decreased the levels of p-AKT and AKT, as well as their transcript levels. While NEK2 inhibition reduced the rates of cleavage and blastocyst formation as well as total cell number, all of these effects were reversed by SC79 co-treatment. The proportions of expanded blastocyst and cell survival and the trophectoderm cell numbers were similarly restored to control levels following combined treatment with SC79. Furthermore, the reduced levels of EdU, EU, and OPP by NEK2 inhibition were completely restored by SC79 co-treatment. NEK2 inhibition had a negative impact on DNA integrity, and this effect was inhibited by SC79 co-treatment. Significance: Together, these results suggest that NEK2 plays a crucial role in porcine embryonic development by regulating the AKT signaling pathway.