Journal
THERIOGENOLOGY
Volume 158, Issue -, Pages 358-367Publisher
ELSEVIER SCIENCE INC
DOI: 10.1016/j.theriogenology.2020.09.031
Keywords
Microcystin-LR; Porcine oocyte; Spindle; Oxidative stress; Apoptosis
Categories
Funding
- National Natural Science Foundation of China [31972759, 31572589]
- Nanjing Agricultural University College Students Innovation and Entrepreneurship Training Special Plan [S20190021]
- National College Students Innovation and Entrepreneurship Training Program [201910307022Z]
- Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD)
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Microcystin-LR (MC-LR), as a well-known hepatotoxin, was recently found to accumulate in gonads and induce a variety of reproductive damages in zebrafish, mice and other model organisms, however, little information is available on whether MC-LR has toxic effects on the mammalian oocytes, especially in livestock species. In this study, the effects of MC-LR on meiotic maturation of porcine oocytes were investigated, and the potential mechanism of MC-LR toxicity was explored. Germinal vesicle (GV)-stage oocytes were exposed to 0, 20, 40 and 60 mu M MC-LR, respectively, during the in vitro maturation for 44 h, and the results showed that the first polar body (PbI) extrusion rate of the oocytes decreased significantly when the MC-LR concentration reached 40 (P < 0.01) or 60 mu M (P < 0.001). After treated with 60 mu M MC-LR for 44 h, a significant higher percentage of the oocytes arrested at anaphase-telophase I (ATI) stage (P < 0.01). Laser scanning confocal results further confirmed that a significantly larger proportion of the 60 mu M MC-LR-treated oocytes exhibited aberrant spindles and misaligned chromosomes, suggesting a failure of spindle assembly and homologous chromosome segregation during the ATI stage. Furthermore, the ROS levels in the 60 mu M MC-LR-exposed oocytes were significantly higher than the control group (P < 0.01), while the expression of antioxidant related genes (SOD1, CAT and GPX) were much lower compared with control group, indicating that oxidative stress was induced and the antioxidant capacity of oocytes was depleted by 60 mu M MC-LR treatment. Additionally, markedly decreased mitochondrial membrane potential (MMP) (P < 0.01) and significantly higher incidence of early apoptosis (P < 0.01) were observed in the 60 mu M MC-LR-treated oocytes, suggesting that MC-LR exposure induced apoptosis in porcine oocytes. Moreover, the protein expression of PP2A was remarkably inhibited, whereas the expression of p53, BAX, Caspase3 and Cleaved-caspase3 were prominently increased in the 60 mu M MC-LR-exposed oocytes. Together, these results suggested that 60 mu M of MC-LR exposure can induce oxidative stress, and lead to aberrant spindles, impaired MMP, and trigger apoptosis, which eventually result in failure of porcine oocyte maturation. (C) 2020 Elsevier Inc. All rights reserved.
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