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农业生物技术学报  2020, Vol. 28 Issue (5): 859-865    DOI: 10.3969/j.issn.1674-7968.2020.05.009
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Effects of MC-LR on in vitro Maturation of Porcine (Sus scrofa) Oocytes
SHI Feng-Yao, LI Wen-Hui, ZHAO Hong-Yu, ZHUANG Rui-Xue, WANG Yang, RUI Rong, JU Shi-Qiang*
College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China
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Abstract  Microcystin-LR (MC-LR) is a widespread toxin produced by Cyanobacteria that pollutes the water environment. Animal studies provide strong evidence of positive associations between microcystins (MCs) exposure and reproductive toxicity, representing a threat to human reproductive health and the biodiversity of wild life. However, little is known about its toxic effects on animal oocytes. This study evaluated the toxic effects of MC-LR on porcine (Sus scrofa) oocyte in vitro maturation. The cumulus-oocyte complexes (COCs) were exposed to 0, 20, 40, 60 μg/mL MC-LR for maturation culture. Immunofluorescent staining coupled with confocal microscopy imaging techniques was used to examine the chromosomes alignment and spindle tubulin distribution of the MC-LR treated oocytes. The results showed that the maturation rate of pig oocytes exposed to MC-LR decreased in a dose-dependent manner. After 44 h in culture, most of the control oocytes had extruded the first polar body and arrested at the M stage in vitro, and the first polar body extrusion rate was 72.81%. However, the rate was significantly reduced to 56.71% (P<0.05) and 45.00% (P<0.01) when treated with 40 and 60 μg/mL of MC-LR, respectively. After MC-LR (60 μg/mL) treatment, the proportion of the oocytes arrested in telophase (T) stage was significantly increased from 10.63% to 45.49% (P<0.001), and the proportion of the oocytes showed obvious defects in homologous chromosome segregation and alignmen was also increased from 11.94% to 44.33% (P<0.001). Further subcellular structure examination results showed that the proportion of oocytes with spindle structure disorder increased significantly after MC-LR treatment. The results of this study indicated that MC-LR had obvious toxic damage to the spindle structure of porcine oocyte, resulting in abnormal alignment of homologous chromosomes and arrestment of meiosis in T stage, eventually resulted in failure of porcine oocyte maturation (P<0.01). The results will provide an experimental basis for further understanding the reproductive toxicity and its mechanism of MC-LR, and also provide experimental reference for effective prevention and treatment of diseases related to the reproductive toxicity of MC-LR
Key wordsMicrocystin-LR (MC-LR)      Oocytes      Meiosis division      Spindle      Chromosome     
Received: 23 October 2019     
ZTFLH:  S857.2  
Corresponding Authors: *jusq@njau.edu.cn   
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SHI Feng-Yao
LI Wen-Hui
ZHAO Hong-Yu
ZHUANG Rui-Xue
WANG Yang
RUI Rong
JU Shi-Qiang
Cite this article:   
SHI Feng-Yao,LI Wen-Hui,ZHAO Hong-Yu, et al. Effects of MC-LR on in vitro Maturation of Porcine (Sus scrofa) Oocytes[J]. 农业生物技术学报, 2020, 28(5): 859-865.
URL:  
http://journal05.magtech.org.cn/Jwk_ny/EN/10.3969/j.issn.1674-7968.2020.05.009     OR     http://journal05.magtech.org.cn/Jwk_ny/EN/Y2020/V28/I5/859
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