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Oocyte maturation plays a vitally important role in porcine reproduction. Regrettably, the quality of oocytes matured in vitro is weaker than that of in vivo matured oocytes. We collected and cultivated porcine cumulus oocyte complexes (COCs) in vitro with phosphoinositide-dependent kinase 1 (PDK1) activator 5-(4-chloro-phenyl)-3-phenyl-pent-2-enoic acid (PS48), whose concentrations were 0, 2, 5, 10 and 20 µM to investigate whether the phosphatidylinositol-3-kinase/protein kinase B (PI3K/Akt) signalling pathway would impact the oocyte quality. The results showed that 10 µM PS48 increased the oocyte proportion of metaphase II (MII) stage and improved the expansion of cumulus cells (CCs). What's more, the activation of PI3K/Akt signalling pathway could regulate the expression of maturation-related genes and proteins. The results of quantitative real-time PCR showed that 10 µM PS48 increased the mRNA and protein levels of Akt and regulated maturation-related genes, including cyclin B1, MOS, BMP15, GDF9, CDC2, mTOR, BAX, BCL2 and caspase-3. The results of Western blot indicated that 10µM PS48 increased the protein abundance of Akt, phosphorylation of Akt Thr308 (p-AktThr308 ) and cyclin B1, but decreased the protein abundance of pro-apoptotic BAX. These results suggested that adding 10 µM PS48 to mature culture medium could promote the maturation of porcine oocytes, potentially through activating the PI3K/Akt signalling pathway.

The phosphatidylinositol -3- kinase (PI3K) signaling pathway is critical for the cell proliferation, apoptosis, metabolism, DNA repair and protein synthesis. Significant effort has focused on elucidating the relationship between PI3K signaling pathway and other nuclear signal transducers; However, little is known about the connection between PI3K signaling pathway and porcine oocyte meiotic maturation. In this study, we investigated the function of PI3K signaling pathway in porcine oocytes. PI3K signaling pathway was important during oocyte maturation. Furthermore, the PI3K signaling pathway inhibitor LY-294002 blocked porcine oocyte maturation, reducing the percentage of oocytes that first polar body (PBI) extrusion. LY-294002 also decreased the expression of oocyte proliferation-related gene PCNA and reduced the mRNA and protein levels of PI3K. What’s more, LY-294002 also decreased other maturation-related genes that are predominantly expressed duringporcine oocyte maturation, including bone morphogenetic protein 15 (BPM15), growth differentiation factor 9 (GDF9), cell division cycle protein 2 (CDC2), phosphatase and tensin homolog (PTEN), CyclinB1, MOS and Akt. LY-294002 treatment decreased the developmental potential of blastocysts following parthenogenetic activation, increased the level of cell apoptosis and reduced the level of cell-cycle. This study revealed that inhibiting the PI3K signaling pathway could reduce in vitro maturation and developmental competence of porcine oocytes, probably by reducing cell cycle arrest and proliferation, promoting the oocyte apoptosis, and altering the expression of other maternal genes.

In somatic cell reprogramming, cells must escape the somatic cell-specific gene expression program to adopt other cell fates. Here, in vitro chemical induction with RepSox generated chemically induced mammary epithelial cells (CiMECs) with milk secreting functions from goat ear fibroblasts (GEFs). Transplanted CiMECs regenerated the normal mammary gland structure with milk-secreting functions in nude mice. Single-cell RNA sequencing revealed that during the reprogramming process, GEFs may sequentially undergo embryonic ectoderm (EE)-like and different MEC developmental states and finally achieve milk secreting functions, bypassing the pluripotent state. Mechanistically, Smad3 upregulation induced by transforming growth factor β (TGFβ) receptor 1 (TGFβR1) downregulation led to GEF reprogramming into CiMECs without other reprogramming factors. The TGFβR1-Smad3 regulatory effects will provide new insight into the TGFβ signaling pathway regulation of somatic cell reprogramming. These findings suggest an innovative strategy for autogenous cell therapy for mammary gland defects and the production of transgenic mammary gland bioreactors.