Anthocyanin stimulates in vitro development of cloned pig embryos by increasing the intracellular glutathione level and inhibiting reactive oxygen species

Abstract

The objective was to examine the nuclear maturation of oocytes, embryonic development after parthenogenetic activation (PA) and somatic cell nuclear transfer (SCNT), and gene expression in SCNT embryos in pigs ( Sus scrofa) when anthocyanin was added to oocytes during maturation and in vitro culture (IVC) of embryos. Immature oocytes were untreated or treated with 0.1 μg/mL anthocyanin during in vitro maturation (IVM). Next, PA and SCNT embryos were produced from oocytes and cultured in medium supplemented with or without 0.1 μg/mL anthocyanin for 7 d. Anthocyanin treatment during IVM did not improve the nuclear maturation of oocytes, but significantly increased intracellular glutathione (GSH) levels and reduced reactive oxygen species (ROS). Oocytes treated with anthocyanin during IVM had higher ( P < 0.05) rates of blastocyst formation after PA (55.7 vs. 44.9 %) and SCNT (32.2 vs. 16.1%) compared to untreated oocytes. In PA and SCNT embryos, anthocyanin treatment during IVM or IVC significantly increased the intracellular GSH level, which led to the reduced ROS level. Somatic cell nuclear transfer embryos derived from anthocyanin-treated oocytes had increased ( P < 0.05) expression of DNMT1, PCNA, FGFR2, and POU5F1 mRNA compared to control embryos. In conclusion, anthocyanin treatment during IVM improved developmental competence of SCNT embryos, most likely by increasing intracellular GSH synthesis, reducing ROS level, and stimulating nuclear reprogramming via increased transcription factor expression.