乙草胺对小鼠卵母细胞体外成熟的影响

doi:10.16431/j.cnki.1671-7236.2022.10.023

Abstract

Abstract: 【Objective】 The aim of study was to explore the effect of acetochlor on the maturation of mouse oocytes in vitro,in order to provide a basic reference for judging the toxicity standard of acetochlor.【Method】 The collected cumulus-oocyte complexes (COCs) were placed in the in vitro maturation solution containing different concentrations (0 (control group),10,50,100,130,200 μmol/L) of acetochlor for 14 h,the first polar body excretion rate of oocytes was calculated.After in vitro fertilization(IVF),the blastocyst rate of IVF embryos was counted,suitable acetochlor concentrations for oocyte culture in vitrowereselected out.After COCs were cultured in mature medium of control group and selected appropriate concentrations acetochlor group for 14 h,the ROS levels of oocyteswere detected by DCFH-DA.The GSH levels of oocytes were detectedby CellTracker^TM blue dye.The mitochondrial membrane potential (MMP) of oocytes was detected by JC-1 staining method.The expression levels of apoptosis-related genes in oocytes were detected by Real-time quantitative PCR.The total number of blastocyst cells of IVF embryos was detected by Hoechst 33342 staining,and the apoptosis rate of blastocyst cells was detected by Fluorescein-dUTP.【Result】 Compared with control group,the first polar body excretion rate of oocytes in 100 and 130 μmol/L acetochlor groups was significantly decreased (P＜0.05),and the blastocyst rate of VIF embryos in 100 and 130 μmol/L acetochlor groups was significantly decreased (P＜0.05),and IVF embryos in 200 μmol/L acetochlor group did not develop to blastocyst stage,therefore,100 and 130 μmol/L acetochlor were selected for follow-up tests.Compared with control group,in 100 and 130 μmol/L acetochlor groups,the level of ROS in oocytes was significantly increased (P＜0.05),the level of GSH and mitochondrial membrane potential were significantly increased (P＜0.05),the relative expression of the pro-apoptotic gene Caspase9 in the oocytes was significantly increased (P＜0.05),while the relative expression of the anti-apoptotic genes Bax and Bcl-2was significantly increased(P＜0.05).In 100 and 130 μmol/L acetochlor groups,the total number of blastocyst cells was significantly decreased (P＜0.05),the blastocyst cell apoptosis rate was significantly increased (P＜0.05).【Conclusion】 Acetochlor reduced the quality of oocytes by increasing ROS and promoting apoptosis of oocytes,thus affecting the developmental potential of oocytes after fertilization.

Key words: acetochlor; mice; oocytes; in vitro maturation; in vitro fertilization; early embryos

CLC Number:

WANG Yu, ZHAO Yuhan, JIAO Anhui, WANG Yuqi, GAO Qingshan. Effects of Acetochlor on in vitro Maturation of Mouse Oocytes[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(10): 3909-3917.

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Effects of Acetochlor on in vitro Maturation of Mouse Oocytes

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