Using cysteine/cystine to overcome oxidative stress in goat oocytes and embryos cultured in vitro.
Assisted reproductive techniques expose gametes to excessive concentrations of reactive oxygen species. The present study aimed to evaluate the effects of oxidative stress on apoptosis in goat oocytes and embryonic development. The results demonstrated that the addition of 100 µM hydrogen peroxide (H2O2) into media produces an oxidative environment during oocyte maturation. The number of cumulus cells positive for terminal deoxynucleotidyl transferase UTP nick end labeling, and the activity of caspase 3 in mature oocytes were increased, compared with the control group (P<0.05). In addition, the expression levels of mitochondrial regulators, including peroxisome proliferator‑activated receptor γ coactivator-1 α (PGC-1α) and nuclear respiratory factor‑1 (NRF‑1) were increased in the oxidative oocytes, compared with those in the control group (P<0.05). The ratio of the proapoptotic gene, B cell lymphoma (Bcl-2)-associated X protein (BAX), to the anti‑apoptotic gene, BCL‑2, was higher in the H2O2 group, compared with the control group (P<0.05). To overcome oxidative stress in oocytes and embryos cultured in vitro, 200 µM cysteine and 200 µM cystine were added to the media, thereby increasing the concentration of intracellular glutathione (GSH) and assisting in maintaining the redox state of the cells. In conclusion, cysteine and cystine reduced the oxygen tension caused by H2O2, thereby providing a novel strategy for optimizing in vitro embryonic development systems.
Jiangsu Livestock Embryo Engineering Laboratory, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, Jiangsu 210095, P.R. China.
Zhengrong
Z
Zhou
Jiangsu Livestock Embryo Engineering Laboratory, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, Jiangsu 210095, P.R. China.
Ruo-Xin
RX
Jia
Jiangsu Livestock Embryo Engineering Laboratory, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, Jiangsu 210095, P.R. China.
Guomin
G
Zhang
Jiangsu Livestock Embryo Engineering Laboratory, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, Jiangsu 210095, P.R. China.
Yongjie
Y
Wan
Jiangsu Livestock Embryo Engineering Laboratory, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, Jiangsu 210095, P.R. China.
Yanli
Y
Zhang
Jiangsu Livestock Embryo Engineering Laboratory, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, Jiangsu 210095, P.R. China.
Yixuan
Y
Fan
Jiangsu Livestock Embryo Engineering Laboratory, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, Jiangsu 210095, P.R. China.
Ziyu
Z
Wang
Department of Histology and Embryology, School of Medicine, Jiangsu University, Zhenjiang, Jiangsu 212013, P.R. China.
Pan
P
Huang
Jiangsu Livestock Embryo Engineering Laboratory, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, Jiangsu 210095, P.R. China.
Feng
F
Wang
D000818:Animals; D015415:Biomarkers; D001755:Blastocyst / Q000187:drug effects / Q000378:metabolism; D053148:Caspase 3 / Q000378:metabolism; D002478:Cells, Cultured; D003545:Cysteine / Q000494:pharmacology*; D003553:Cystine / Q000494:pharmacology*; D004622:Embryo, Mammalian / Q000187:drug effects* / Q000378:metabolism*; D047108:Embryonic Development / Q000187:drug effects / Q000235:genetics; D005260:Female; D015870:Gene Expression; D020869:Gene Expression Profiling; D005978:Glutathione / Q000378:metabolism; D006041:Goats; D006861:Hydrogen Peroxide / Q000494:pharmacology; D066298:In Vitro Techniques; D009865:Oocytes / Q000187:drug effects* / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects; D017382:Reactive Oxygen Species / Q000378:metabolism
D015415:Biomarkers; D017382:Reactive Oxygen Species; D003553:Cystine; D006861:Hydrogen Peroxide; D053148:Caspase 3; D005978:Glutathione; D003545:Cysteine