2-氨基乙基联苯基硼酸酯对玻璃化冷冻牛成熟卵母细胞发育能力的影响

doi:10.16431/j.cnki.1671-7236.2022.07.025

Abstract

Abstract: 【Objective】 The study was aimed to investigate the effects of endoplasmic reticulum IP3 receptor (IP3R) inhibitor 2-aminoethyl diphenylborinate (2-APB) on the developmental capacity of frozen-thawed bovine MⅡoocytes.【Method】 Oocytes matured in vitro for 24 h were digested with 0.1% hyaluronidase to remove cumulus cells,and were randomly divided into control group and 2-APB group,the open pulled straw (OPS) method was used for vitrification.The control group was vitrified directly,2-APB group was pretreated with 10 μmol/L 2-APB for 10 min before vitrification.Oocytes were thawed after 2 d,the survival rate of oocytes after thawed (0 h) and warmed for 2 h were counted.After recovering for 2 h, cortical granules (CG)were labeled with FITC-PNA fluorescent probe to detect distribution,intracellular reactive oxygen species (ROS) and glutathione (GSH) contents were deteced by 2',7'- DCHFDA and Cell Tracker Blue CMF2HC respectively.The non-vitrified oocytes (Fresh group)matured in vitro for 24 h were parthenogenetic activated as well as those of the control and 2-APB groups,the cleavage and blastocyst rates were counted at 2 and 7 d,respectively.【Result】 Compared with control group,the survival rate of frozen-thawed oocytes at 0 and 2 h in 2-APB group was significantly increased (P＜0.05),the proportion of CG distributed in cortical area in 2-APB group was significantly increased (P＜0.05),the proportion of CG injury type was significantly decreased (P＜0.05),the content of GSH was significantly increased (P＜0.05),and the content of ROS was significantly decreased (P＜0.05),the cleavage and blastocyst rates of parthenogenetic activated embryos in 2-APB group were significantly increased (P＜0.05),and there was no significant difference with the Fresh group (P＞0.05).【Conclusion】 Pretreatment of bovine MⅡ oocytes with 2-APB before vitrification could increase GSH content,decrease the proportion of CG injury type and ROS content,improve the frozen-thawed oocyte quality and early embryo developmental capacity.

Key words: vitrification; 2-aminoethyl diphenylborinate; oocyte; development capacity; bovine

CLC Number:

S823

MO Xianhong, YUE Kaiping, SUN Liyao, LI Bing, ZHAO Bing, GUO Cheng, XU Zhenjun. Effect of 2-APB on Development Capacity of Vitrified Bovine Mature Oocytes[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(7): 2669-2676.

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Effect of 2-APB on Development Capacity of Vitrified Bovine Mature Oocytes

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