Improvement of in vitro-produced bovine embryo treated with coagulansin-A under heat-stressed condition

Reproduction. 2017 Apr;153(4):421-431. doi: 10.1530/REP-16-0530. Epub 2017 Jan 9.

Abstract

Heat stress has large effects on reproduction including conception rate in cattle. In this study, we examined the effects of coagulansin-A (coa-A), a steroidal lactone, on acquired thermo tolerance during in vitro production of bovine embryos. Oocytes were incubated in in vitro maturation (IVM) media with or without coa-A at two different temperatures, 40.5˚C and 42˚C, for 20 h. The treatment of coa-A significantly improved blastocyst development only at 40.5˚C (P < 0.05). Interestingly, immunofluorescence analysis demonstrated that coa-A induced heat shock protein 70 (HSP70) and phosphatidylinositol-3-kinase (PI3K), but significantly attenuated nuclear factor kappa B (NF-κB) and cyclooxygenase-2 (COX2). To determine the expression patterns of related genes at the transcription level, qRT-PCR was performed. Expression of HSP70 and PI3K was elevated, whereas expression of NF-κB, COX2 and inducible nitric oxide synthase (iNOS) was significantly (P < 0.05) downregulated in the coa-A-treated group compared with the control group. Moreover, pro-apoptotic genes were downregulated, and antiapoptic genes were upregulated in the coa-A group. We also counted the total cell number and apoptotic nuclei at the blastocyst and found that more cell numbers (143.1 ± 1.5) and less apoptotic damages (6.4 ± 0.5) in the coa-A treatment group comparing to control group (131.4 ± 2.0 and 10.8 ± 0.5), indicating the enhanced embryo quality. In conclusion, our results demonstrate that the coa-A not only improved the blastocyst development in vitro but also increased their resistance to heat stress condition through induction of HSP70/PI3K.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cattle
  • Embryo, Mammalian / cytology*
  • Embryo, Mammalian / drug effects
  • Embryo, Mammalian / metabolism
  • Embryonic Development / drug effects*
  • Female
  • Fertilization in Vitro / drug effects
  • Gene Expression Regulation, Developmental / drug effects*
  • HSP70 Heat-Shock Proteins / metabolism
  • Heat Stress Disorders / prevention & control*
  • Hot Temperature / adverse effects
  • Oocytes / cytology
  • Oocytes / drug effects
  • Oocytes / metabolism
  • Withanolides / chemistry
  • Withanolides / pharmacology*

Substances

  • HSP70 Heat-Shock Proteins
  • Withanolides