Melatonin reduces apoptotic cells, SOD2 and HSPB1 and improves the in vitro production and quality of bovine blastocysts

Abstract

Effects of adding different concentrations of melatonin (10 , 10 and 10 M) to maturation (Experiment 1; Control, IVM + 10 , IVM + 10 , IVM + 10 ) and culture media (Experiment 2; Control, IVC + 10 , IVC + 10 , IVC + 10 ) were evaluated on in vitro bovine embryonic development. The optimal concentration of melatonin (10 M) from Experiments 1-2 was tested in both maturation and/or culture media of Experiment 3 (Control, IVM + 10 , IVC + 10 , IVM/IVC + 10 ). In Experiment 1, maturated oocytes from Control and IVM + 10 treatments showed increased glutathione content, mitochondrial membrane potential and percentage of Grade I blastocysts (40.6% and 43%, respectively). In Experiment 2, an increase in the percentage of Grade I blastocysts was detected in IVC + 10 (43.5%; 56.7%) and IVC + 10 (47.4%; 57.4%). Moreover, a lower number and percentage of apoptotic cells in blastocysts were observed in the IVC + 10 group compared to Control (3.8 ± 0.6; 3.6% versus 6.1 ± 0.6; 5.3%). In Experiment 3, the IVC + 10 treatment increased percentage of Grade I blastocysts with a lower number of apoptotic cells compared to IVM/IVC + 10 group (52.6%; 3.0 ± 0.5 versus 46.0%; 5.4 ± 1.0). The IVC + 10 treatment also had a higher mRNA expression of antioxidant gene (SOD2) compared to the Control, as well as the heat shock protein (HSPB1) compared to the IVM + 10 . Reactive oxygen species production was greater in the IVM/IVC + 10 treatment group. In conclusion, the 10 M concentration of melatonin and the in vitro production phase in which it is used directly affected embryonic development and quality.

α-Tocopherol modifies the expression of genes related to oxidative stress and apoptosis during in vitro maturation and enhances the developmental competence of rabbit oocytes

“…In fact, in mice and human MII oocytes, no CATtranscripts have been detected (El Mouatassim et al 1999), and downregulation of SOD2 mRNA transcripts is associated with higher oocyte ´ developmental competence (Arias-Alvarez et al 2017). In contrast, it has been reported that oocytes (Khalil et al 2013) and embryos (Marques et al 2018) with a better antioxidant defence show higher levels of endogenous antioxidant enzymes. These apparent discrepancies could be due to the addition of different types of antioxidants to the IVM medium and differ ences between species.…”

Effects of melatonin on production of reactive oxygen species and developmental competence of bovine oocytes exposed to heat shock and oxidative stress duringin vitromaturation

“…Melatonin affects cellular function by interacting with membrane and nuclear receptors and by functioning as an antioxidant (Tan et al , 2002; Mayo et al , 2017). Melatonin improved oocyte maturation in the pig (Shi et al , 2009), sheep (Tian et al , 2017) and bovine (El-Raey et al , 2011; Tian et al , 2014; Marques et al , 2018).…”

Use of trichostatin A alters the expression of HDAC3 and KAT2 and improves in vitro development of bovine embryos cloned using less methylated mesenchymal stem cells

“…Total RNA was extracted from three pools of four embryos for each treatment using the PicoPure ® Kit (Applied Biosystems) according to manufacturer's instructions. cDNA syntheses and PCR were performed according to Marques et al ().…”