Iloprost supports early development of in vitro-produced porcine embryos through activation of the phosphatidylinositol 3-kinase/AKT signalling pathway

Reprod Fertil Dev. 2017 Jul;29(7):1306-1318. doi: 10.1071/RD15391.

Abstract

Despite evidence of the presence of prostaglandin (PG) I2 in mammalian oviducts, its role in early development of in vitro-produced (IVP) embryos is largely unknown. Thus, in the present study we examined the effects of iloprost, a PGI2 analogue, on the in vitro developmental competence of early porcine embryos and the underlying mechanism(s). To examine the effects of iloprost on the development rate of IVF embryos, iloprost was added to the in vitro culture (IVC) medium and cultured for 6 days. Supplementation of the IVC medium with iloprost significantly improved developmental parameters, such as blastocyst formation rate, the trophectoderm:inner cell mass ratio and cell survival in IVF and parthenogenetically activated (PA) embryos. In addition, post-blastulation development into the expanded blastocyst stage was improved in iloprost-treated groups compared with controls. Interestingly, the phosphatidylinositol 3-kinase (PI3K)/AKT signalling pathway was significantly activated by iloprost supplementation in a concentration-dependent manner (10-1000nM), and the beneficial effects of iloprost on the early development of porcine IVF and PA embryos was completely ablated by treatment with 2.5μM wortmannin, a PI3K/AKT signalling inhibitor. Importantly, expression of the PI3K/AKT signalling pathway was significantly reduced in somatic cell nuclear transfer (SCNT) compared with IVF embryos, and iloprost supported the early development of SCNT embryos, as was the case for IVF and PA embryos, suggesting a consistent effect of iloprost on the IVC of IVP porcine embryos. Together, these results indicate that iloprost can be a useful IVC supplement for production of IVP early porcine embryos with high developmental competence.

MeSH terms

  • Androstadienes / pharmacology
  • Animals
  • Culture Media
  • Embryo Culture Techniques / methods
  • Embryo Culture Techniques / veterinary
  • Embryonic Development / drug effects*
  • Embryonic Development / physiology
  • Epoprostenol / analogs & derivatives
  • Epoprostenol / pharmacology
  • Female
  • Fertilization in Vitro / methods
  • Fertilization in Vitro / veterinary
  • Iloprost / pharmacology*
  • Models, Biological
  • Nuclear Transfer Techniques / veterinary
  • Parthenogenesis
  • Phosphatidylinositol 3-Kinases / metabolism*
  • Proto-Oncogene Proteins c-akt / metabolism*
  • Signal Transduction / drug effects
  • Sus scrofa / embryology*
  • Sus scrofa / metabolism*
  • Wortmannin

Substances

  • Androstadienes
  • Culture Media
  • Epoprostenol
  • Phosphatidylinositol 3-Kinases
  • Proto-Oncogene Proteins c-akt
  • Iloprost
  • Wortmannin