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首页> 外文期刊>Veterinary World >The effect of vitrification after warming on the expressions of p38, CDK1, and cyclin B in immature goat oocytes followed by in vitro maturation
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The effect of vitrification after warming on the expressions of p38, CDK1, and cyclin B in immature goat oocytes followed by in vitro maturation

机译：在不成熟的山羊卵母细胞中升温后玻璃化玻璃化在不成熟的羊毛卵母细胞中的效果，然后进行体外成熟

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Background and Aim: The combination of vitrification techniques and in vitro maturation can reduce oocyte competence. Mitogen-activated protein kinase and maturation-promoting factor are significant in oocyte meiotic maturation regulation. This study aimed to analyze vitrification's effect, after warming followed by in vitro maturation, on the expressions of protein 38 (p38), cyclin-dependent kinase 1 (CDK1), and cyclin B and oocyte maturation level. Materials and Methods: Immature goat oocytes were soaked in vitrification and warming solutions. The procedure was followed by in vitro maturation and in vitro maturation without post-warming vitrification as a control. These oocytes, along with their cumulus, were vitrified using hemistraw in liquid nitrogen. Oocyte maturation was carried out in a maturation medium that was added with 10 μg/mL of FSH, 10 μg/mL of LH, and 1 μg/mL E 2 for 22 h. The expressions of p38, CDK1, and cyclin B were observed using immunocytochemical methods, which were assessed semiquantitatively according to the modified Remmele method. The oocyte maturation level was observed using the aceto-orcein staining method based on the achievement of chromosomes up to the metaphase II stage and/or the formation of the polar body I. Results: p38 expression in vitrified oocytes after warming, followed by in vitro maturation, increased insignificantly (p≥0.05), with the acquisition of 3.91±2.69 and 2.69±0.50 in the control oocytes. CDK1 expression in vitrified oocytes decreased significantly (p≤0.05) after warming, followed by in vitro maturation, with the acquisition of 2.73±1.24 and 7.27±4.39 in the control oocytes. Cyclin B expression in vitrified oocytes decreased insignificantly (p≥0.05) after warming, followed by in vitro maturation, with the acquisition of 3.09±1.4 and 4.18±2.61 in the control oocytes. The proportion of vitrified oocyte maturation levels after warming, followed by in vitro maturation, decreased significantly (p≤0.05), with the acquisition of 45.45% and 77.27% in the control oocytes. Conclusion: This study concluded that vitrification after warming resulted in an insignificant increase in p38 expression, a significant decrease in CDK1 expression, an insignificant decrease in cyclin B expression, and a significant reduction in oocyte maturation levels.
机译：背景和目的：玻璃化技术和体外成熟的组合可以降低卵母细胞能力。促丝糖活性蛋白激酶和成熟促进因子在卵母细胞的降低成熟调节中是显着的。本研究旨在在玻璃蛋白38（p38），细胞周期蛋白依赖性激酶1（CDK1）和细胞周期蛋白B和卵卷B和卵母细胞成熟水平上进行玻璃化后分析玻璃化效果。材料和方法：将未成熟的山羊卵母细胞浸泡在玻璃化和升温溶液中。然后在体外成熟和体外成熟的过程，而不会使玻璃化剂作为对照。这些卵母细胞以及它们的积积菌在液氮中玻璃化。卵母细胞成熟在成熟培养基中进行，所述成熟培养基中加入10μg/ ml的FSH，10μg/ ml的LH和1μg/ ml E 2 22小时。使用免疫细胞化学方法观察P38，CDK1和细胞周期蛋白B的表达，其根据改性的雷梅尔法来分半定量评估。使用基于染色体的染色体染色方法和/或极性体I的形成，观察卵母细胞效果水平。结果：升温后玻璃化卵母细胞的P38表达，然后进行体外成熟，无关紧要（P≥0.05），获取控制卵母细胞中的3.91±2.69和2.69±0.50。在加热后，玻璃化卵母细胞中的CDK1表达显着降低（p≤0.05），然后进行体外成熟，在对照卵母细胞中采集2.73±1.24和7.27±4.39。在加热后，玻璃化卵母细胞中的细胞周期蛋白B表达在升温后（p≥0.05）减少，然后在体外成熟后，采集3.09±1.4和4.18±2.61，在对照卵母细胞中。玻璃化卵母细胞成熟水平的比例升温，然后体外成熟，显着降低（p≤0.05），获取控制卵母细胞中的45.45％和77.27％。结论：本研究得出结论，玻璃化在升温后导致P38表达的微不足道，CDK1表达的显着降低，细胞周期蛋白B表达的微不足道降低，以及卵母细胞成熟水平的显着降低。

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