卵泡细胞中的基因表达水平与牛卵母细胞的发育能力相关

Gene expression levels in cumulus cells are correlated with developmental competence of bovine oocytes

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THERIOGENOLOGY

影响因子:2.5

分区:农林科学2区 / 兽医学1区生殖生物学3区

发表日期:2025 Jan 01

作者: Mayuko Anazawa, Shiori Ashibe, Yoshikazu Nagao

DOI: 10.1016/j.theriogenology.2024.09.031

摘要

哺乳动物胚胎的体外培养产生受阻，主要原因是许多胚胎无法发育到囊胚阶段。即使选择形态良好的卵泡-卵母细胞复合体（COCs）用于培养，也会出现此问题。由于卵泡细胞对卵母细胞的成熟和随后的胚胎发育至关重要，本研究比较了在体外能发展到囊胚的COCs与未能完成该过程的COCs中卵泡细胞的基因表达模式。取自牛的COCs中的卵泡细胞，评估其发育能力，通过检测细胞分裂及发育至囊胚的情况。对收集的卵泡细胞进行mRNA水平检测，包括促卵泡刺激素受体（FSHR）、胰岛素样生长因子-1受体（IGF-1R）、抗米勒管激素（AMH）、AMH受体II（AMHRII）、表皮生长因子受体（EGFR）、雌激素受体β（ERβ）、B细胞淋巴瘤/白血病-2相关X（Bax）及半胱天冬酶-3（Caspase-3）。结果显示，发育至囊胚的COCs，其卵泡细胞中FSHR、IGF-1R、AMH和EGFR的表达水平较高，而Bax与Caspase-3的表达较低。发育至囊胚的COCs的卵泡细胞中，FSHR与IGF-1R表达呈正相关（r=0.59），ERβ与EGFR表达也呈正相关（r=0.43）。这些发现表明，卵泡细胞中的基因表达水平与牛卵母细胞的发育能力相关。测定卵泡细胞中的基因表达提供了一种非侵入性预测卵母细胞发育能力的方法。

Abstract

The generation of mammalian embryos by in vitro culture is hampered by the failure of many of the embryos to develop to the blastocyst stage. This problem occurs even when cumulus-oocyte complexes (COCs) with good morphology are visually selected and used for culture. Because cumulus cells are important for oocyte maturation and subsequent embryo development, here we compared gene expression patterns in cumulus cells of COCs that developed in vitro to the blastocyst stage with those of COCs that failed to develop. Cumulus cells were aspirated from bovine COCs selected for in vitro culture. Oocyte developmental competence was evaluated by screening for cleavage and development to the blastocyst stage. The collected cumulus cells were used to quantify mRNA levels of FSH receptor (FSHR), insulin-like growth factor-1 receptor (IGF-1R), anti-Müllerian hormone (AMH), AMH receptor II (AMHRII), epidermal growth factor receptor (EGFR), estrogen receptor β (ERβ), B cell lymphoma/leukemia-2 associated X (Bax), and cysteine-aspartic acid protease-3 (Caspase-3). We found that the expression levels of FSHR, IGF-1R, AMH, and EGFR were higher in cumulus cells from COCs that developed to blastocysts as compared with those that failed to develop, whereas expression levels of Bax and Caspase-3 were lower in cumulus cells of COCs that matured to the blastocyst stage. Positive correlations were found between FSHR and IGF-1R expression (r = 0.59) and between ERβ and EGFR expression (r = 0.43) in cumulus cells from COCs that developed to the blastocyst stage. Our findings indicate that gene expression levels in cumulus cells are correlated with the developmental competence of bovine oocytes. Measurement of gene expression in cumulus cells therefore offers a non-invasive means of predicting oocyte developmental competence.