Melatonin reduces apoptotic cells,SOD2 and HSPB1 and improves the in vitro production and quality of bovine blastocysts |
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| Authors: | TC Marques EC da Silva Santos TO Diesel LO Leme CF Martins MAN Dode BG Alves FPH Costa EB de Oliveira ML Gambarini |
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| Affiliation: | 1. Center for Studies and Research in Animal Reproductive Biology, College of Veterinary and Animal Science, Federal University of Goiás, Goiania, GO, Brazil;2. Embrapa Genetic Resources and Biotechnology, Laboratory of Animal Reproduction, Brasília, DF, Brazil;3. Center of Animal Production Systems, Embrapa Cerrados, Brasília, DF, Brazil;4. Laboratory of Manipulation of Oocytes and Preantral Follicles, Faculty of Veterinary, State University of Ceará, Fortaleza, CE, Brazil |
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| Abstract: | Effects of adding different concentrations of melatonin (10?7, 10?9 and 10?11 M) to maturation (Experiment 1; Control, IVM + 10?7, IVM + 10?9, IVM + 10?11) and culture media (Experiment 2; Control, IVC + 10?7, IVC + 10?9, IVC + 10?11) were evaluated on in vitro bovine embryonic development. The optimal concentration of melatonin (10?9 M) from Experiments 1–2 was tested in both maturation and/or culture media of Experiment 3 (Control, IVM + 10?9, IVC + 10?9, IVM /IVC + 10?9). In Experiment 1, maturated oocytes from Control and IVM + 10?9 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?7 (43.5%; 56.7%) and IVC + 10?9 (47.4%; 57.4%). Moreover, a lower number and percentage of apoptotic cells in blastocysts were observed in the IVC + 10?9 group compared to Control (3.8 ± 0.6; 3.6% versus 6.1 ± 0.6; 5.3%). In Experiment 3, the IVC + 10?9 treatment increased percentage of Grade I blastocysts with a lower number of apoptotic cells compared to IVM /IVC + 10?9 group (52.6%; 3.0 ± 0.5 versus 46.0%; 5.4 ± 1.0). The IVC + 10?9 treatment also had a higher mRNA expression of antioxidant gene (SOD 2) compared to the Control, as well as the heat shock protein (HSPB 1) compared to the IVM + 10?9. Reactive oxygen species production was greater in the IVM /IVC + 10?9 treatment group. In conclusion, the 10?9 M concentration of melatonin and the in vitro production phase in which it is used directly affected embryonic development and quality. |
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| Keywords: | antioxidants cellular apoptosis embryonic development melatonin reactive oxygen species SOD2 |
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