Fibronectin 1 supports oocyte in vitro maturation in pigs.

Oocyte in vitro maturation (IVM) based on the follicular fluid (FF) environment can exploit untapped resources, however, what FF factors regulate oocyte maturation remains unclear. This work demonstrated that serum and FF significantly promoted oocyte polar body extrusion (PBE) and subsequent embryo development, and FF was especially effective. Fibronectin 1 (FN1) was predicted as one potential candidate to regulate oocyte maturation by proteomics. FN1 transcription obviously decreased, and the protein expression significantly increased and migrated to plasma membrane or even outside during oocyte IVM. Treatment with 10 ng/mL FN1 significantly improved oocyte PBE rate. FN1 significantly upregulated the percentage of regular spindle morphology, downregulated the γ-H2AX level, decreased the levels of ROS and apoptosis, and increased GSH and mitochondrion contents by ameliorating the expression of corresponding genes. Moreover, FN1 significantly increased the p-PI3K level to enhance the activation of PI3K signaling pathway. In conclusion, this study discovers and confirms that FN1 is one factor in FF that significantly enhances oocyte maturation, and the underlying mechanism is that FN1 ameliorates oocyte nuclear and cytoplasmic maturation by promoting the activation of PI3K signaling pathway.

Astaxanthin enhances the development of bovine cloned embryos by inhibiting apoptosis and improving DNA methylation reprogramming of pluripotency genes.

Low cloning efficiency limits the wide application of somatic cell nuclear transfer technology. Apoptosis and incomplete DNA methylation reprogramming of pluripotency genes are considered as the main causes for low cloning efficiency. Astaxanthin (AST), a powerfully antioxidative and antiapoptotic carotenoid, is recently shown to improve the development of early embryos, however, the potential role of AST during the development of cloned embryos remains unclear. This study displayed that treating cloned embryos with AST significantly increased the blastocyst rate and total blastocyst cell number in a concentration dependent manner, and also alleviated the damage of H2O2 to the development of cloned embryos. In addition, compared with the control group, AST significantly reduced the apoptotic cell number and rate in cloned blastocysts, and the significantly upregulated expression of anti-apoptotic gene Bcl2l1 and antioxidative genes (Sod1 and Gpx4) and downregulated transcription of pro-apoptotic genes (Bax, P53 and Caspase3) were observed in the AST group. Moreover, AST treatment facilitated DNA demethylation of pluripotency genes (Pou5f1, Nanog and Sox2), in accompany with the improved transcription levels of DNA methylation reprogramming genes (Tet1, Tet3, Dnmt1, Dnmt3a and Dnmt3b) in cloned embryos, and then, the significantly upregulated expression levels of embryo development related genes including Pou5f1, Nanog, Sox2 and Cdx2 were observed in comparison with the control group. In conclusion, these results revealed that astaxanthin enhanced the developmental potential of bovine cloned embryos by inhibiting apoptosis and improving DNA methylation reprogramming of pluripotency genes, and provided a promising approach to improve cloning efficiency.

Astaxanthin protects oocyte maturation against cypermethrin-induced defects in pigs.

Cypermethrin (CYP), a pyrethroid insecticide, exerts the detrimental effect on the reproductive system, while astaxanthin (AST), a xanthophyll carotenoid, possesses the powerful antioxidant property and can protect oocyte maturation. However, the toxicity of CYP and the protective role of AST against CYP during oocyte maturation remain unclear. Here, porcine oocytes were applied to investigate the potential effects and underlying mechanisms of CYP and AST during oocyte maturation. This work demonstrated that CYP significantly decreased oocyte maturation rate and subsequent embryo development in a dose-dependent manner (P < 0.05). And, CYP obviously induced the overproduction of reactive oxygen species and the reduction of glutathione content by downregulating the expression of redox genes in oocytes (P < 0.05). Moreover, CYP significantly caused oocyte DNA damage and disturbed the function of endoplasmic reticulum by altering the transcription of DNA damage repair and endoplasmic reticulum stress related genes (P < 0.05). Whereas CYP-exposed oocytes were treated with AST, these defects caused by CYP were significantly ameliorated (P < 0.05). In conclusion, this study demonstrated that CYP exerted the toxic effect on porcine oocytes, while AST effectively alleviated CYP-induced defects. This work provides a potential strategy to prevent pesticide toxicity and protect oocyte maturation in mammalian reproduction.