Increased in vitro production of bovine embryos resulting from oocyte maturation in the presence of triciribine, a specific inhibitor of AKT

IF 2.4 2区 农林科学 Q3 REPRODUCTIVE BIOLOGY Theriogenology Pub Date : 2024-10-24 DOI:10.1016/j.theriogenology.2024.10.024
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Abstract

The aim of this study was to evaluate the effect of different concentrations of triciribine, a selective Akt inhibitor, on various aspects of oocyte maturation and on the IVF of bovine embryos. Cumulus-oocyte complexes (COCs) were matured in vitro in medium supplemented with: 0 (control), 1, 5, 10, and 20 μM of triciribine. The nuclear maturation was assessed by staining with acetic orcein, while the cytoplasmic maturation was evaluated by mitochondrial (MitoTracker® Red CMXRos) and lipid droplets distribution (LipidTOX). COCs were fertilized in vitro and cultured for nine days. Cleavage rates, blastocyst production, and hatching rates were determined on days three, seven, and nine of in vitro culture, respectively. Oocytes from COCs treated with 1 μM of triciribine were stained at 3, 6, and 9 h of IVM to determine the inhibitor's involvement in germinal vesicle breakdown. Analysis of variance (ANOVA) of the data was performed and the means were compared using the SNK test at a 5 % significance level. Exposure of COCs to 1, 5, and 10 μM of triciribine did not alter the number of matured oocytes (P < 0.05), a concentration of 20 μM reduced the number of oocytes in MII with a consequent increase in oocytes in MI (P < 0.05). This concentration markedly reduced the number of oocytes with peripheral cortical granules and the rates of cleavage and blastocysts (P < 0.05). On the other hand, when COCs were matured in the presence of 1 μM, there was an increase in the blastocyst rate (P < 0.05), but without altering the timing of meiosis resumption (P < 0.05). It is concluded that the Akt pathway participates in the nuclear and cytoplasmic events of in vitro maturation of bovine oocytes, but through mechanisms that do not interfere with germinal vesicle breakdown. Modulation of Akt activity in bovine COCs during IVM with 1 μM of triciribine increases the in vitro production of bovine embryos.
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在特异性 AKT 抑制剂曲克来宾的作用下,卵母细胞成熟所产生的牛胚胎体外产量增加。
本研究旨在评估不同浓度的选择性 Akt 抑制剂曲克芦丁对卵母细胞成熟和牛胚胎体外受精各方面的影响。卵丘-卵母细胞复合体(COCs)在体外成熟的培养基中添加了以下添加剂:0(对照组)、1、5、10 和 20 μM 的三尖杉酯碱。核成熟度通过乙酸orcein染色进行评估,而细胞质成熟度则通过线粒体(MitoTracker® Red CMXRos)和脂滴分布(LipidTOX)进行评估。COC 体外受精并培养 9 天。分别在体外培养的第三天、第七天和第九天测定卵裂率、囊胚生成率和孵化率。用 1 μM 三尖杉酯碱处理的 COC 卵母细胞在体外培养 3、6 和 9 小时时进行染色,以确定抑制剂是否参与了生殖囊的破裂。对数据进行方差分析(ANOVA),并在 5 % 的显著性水平上使用 SNK 检验比较平均值。将 COC 暴露于 1、5 和 10 μM 的曲克瑞滨不会改变成熟卵母细胞的数量(P<0.05)。
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来源期刊
Theriogenology
Theriogenology 农林科学-生殖生物学
CiteScore
5.50
自引率
14.30%
发文量
387
审稿时长
72 days
期刊介绍: Theriogenology provides an international forum for researchers, clinicians, and industry professionals in animal reproductive biology. This acclaimed journal publishes articles on a wide range of topics in reproductive and developmental biology, of domestic mammal, avian, and aquatic species as well as wild species which are the object of veterinary care in research or conservation programs.
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