This study was designed not only to measure the effect of delipation on the developmental viability of pig parthenogenetically activated (PA) embryos, but also to evaluate the changes of mitochondria DNA (mtDNA), reactive oxygen species (ROS) level, adenosine triphosphate (ATP) content and gene (Acsl3, Acadsb, Acaa2, Glut1) expression level at different stages after delipation. Results showed that no effect was observed on the cleavage ability, but significant lower blastocyst rate was obtained in delipated embryos. Copy number of mtDNA decreased gradually from MII to four-cell stages and subsequently kept consistent with blastocyst stage both in delipated and control embryos, but the copy number of mtDNA in delipated embryos was similar to that in the control groups no matter at which developmental stage was observed. Both in delipated and control embryos, ATP content progressive decreased from one-cell to blastocyst stages, while just at one-cell stage, a significant decrease of ATP level was observed in delipated embryos compared with that of control. The level of ROS increased obviously after delipation at cleavage stage, but no difference was seen at blastocyst stage. Finally, the expression level of genes related to fatty acids beta-oxidation (Acadsb and Acaa2) was decreased, while the expression level of genes related to glucose metabolism (Glut 1) was upregulated after delipation. In conclusion, the reduction of lipids in pig oocytes will affect the developmental competence of pig PA embryos by disturbed energy metabolism and ROS stress.

ABSTRACT. In Brassicaceae, a self-incompatibility (SI) system mediates pollen-pistil interactions. Self-pollen could be recognized and rejected by incompatible pistils. Several components involved in the SI response have been determined, including S-locus receptor kinase (SRK), S-locus cysteine-rich protein/S-locus protein 11, and arm repeatcontaining protein 1 (ARC1). However, the components involved in the SI system of Brassicaceae are not fully understood. Here, we detected expression patterns of 24 SI-related genes in non-heading Chinese cabbage (Brassica campestris ssp chinensis Makino) after compatible and incompatible pollination, and potential interaction relationships of these genes were predicted. SRK and ARC1 transcripts increased initially 0.25 h after incompatible pollination, while kinase-associated protein phosphatase had an expression pattern that was opposite that of SRK transcripts during self-pollination. Plant U-box 8 was not required in the SI response of non-heading Chinese cabbage. Our results showed that the SI signal of non-heading Chinese cabbage could occur within 0.25 h after self-pollination. The hypothetical interaction relationships indicated that plastid-lipid-associated protein and malate dehydrogenase could be negatively regulated by chaperonin 10, glutathione transferase, cytidylate kinase/uridylate kinase, and methionine synthase by indirect interactions. Our findings could be helpful to better understand potential roles of these components in the SI system of non-heading Chinese cabbage.

Proteins in the LIPIN family play key roles in lipid synthesis mainly on triacylglycerol (TAG) biosynthesis, and they also act as transcriptional coactivators to regulate the expression of genes involved in lipid metabolism with other nuclear factors. Hence, this study was designed to investigate LIPIN1 in pig oocytes and embryos by the delipidation. After delipidation, the content of lipids (LDs) and TAG in MII oocyte was significantly reduced; however, a similar increasing tendency of TAG was shown during embryos development. Subsequently, the expression of genes related to TAG biosynthesis including GPAT1, AGPAT1, AGPAT2, LIPIN1, DGAT and the nuclear factors interacted with LIPIN1 including PPARα and PPARγ was investigated. It is obvious that DGAT and GPAT1, and LIPIN1 increased significantly after delipidation at 1-cell and 4-cell stage, and the expression of PPARα and PPARγ also increased at 4-cell stage. By immunofluorescence staining and Western blots, LIPIN1 was found to exhibit a dynamic localization pattern and gradually increase with the development of delipated embryo. In the early developmental stages (1-, 2- and 4-cell stages), it was distributed over the cortical layer. But at the blastocyst stage, a homogeneous distribution of LIPIN1 was observed in cytoplasm. At 2-cell stage, the expression of PPARα decreased when LIPIN1 was interfered by small interfering RNA, but PPARγ has no significant difference. Therefore, in this study, we find after delipidation, the content of TAG and LIPIN1 will gradually increase during embryo development and nuclear factor PPARα and PPARγ can also be affected by delipidation. The interaction of LIPIN1 and PPARα exists in porcine embryo.

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