Division of Infertility and Reproductive Endocrinology, Department of Gynecology, Obstetrics and Gynecological Oncology, Poznan University of Medical Sciences, Poznan 60‑535, Poland.
Department of Anatomy, Poznan University of Medical Sciences, Poznan 60‑781, Poland.
Mol Med Rep. 2020 Mar;21(3):1537-1551. doi: 10.3892/mmr.2020.10963. Epub 2020 Jan 27.
Oocyte maturation is essential for proper fertilization, embryo implantation and early development. While the physiological conditions of these processes are relatively well‑known, its exact molecular mechanisms remain widely undiscovered. Oocyte growth, differentiation and maturation are therefore the subject of scientific debate. Precious literature has indicated that the oocyte itself serves a regulatory role in the mechanisms underlying these processes. Hence, the present study performed expression microarrays to analyze the complete transcriptome of porcine oocytes during their in vitro maturation (IVM). Pig material was used for experimentation, as it possesses similarities to the reproductive processes and general genetic proximities of Sus scrofa to human. Oocytes, isolated from the ovaries of slaughtered animals were assessed via the Brilliant Cresyl Blue test and directed to IVM. A number of oocytes were left to be analyzed as the 'before IVM' group. Oocyte mRNA was isolated and used for microarray analysis, which was subsequently validated via RT‑qPCR. The current study particularly focused on genes belonging to 'positive regulation of transcription, DNA‑dependent', 'positive regulation of gene expression', 'positive regulation of macromolecule metabolic process' and 'positive regulation of transcription from RNA polymerase II promoter' ontologies. FOS, VEGFA, ESR1, AR, CCND2, EGR2, ENDRA, GJA1, INHBA, IHH, INSR, APP, WWTR1, SMARCA1, NFAT5, SMAD4, MAP3K1, EGR1, RORA, ECE1, NR5A1, KIT, IKZF2, MEF2C, SH3D19, MITF and PSMB4 were all determined to be significantly altered (fold change, >|2|; P<0.05) among these groups, with their downregulation being observed after IVM. Genes with the most altered expressions were analyzed and considered to be potential markers of maturation associated with transcription regulation and macromolecule metabolism process.
卵母细胞成熟对于正常受精、胚胎着床和早期发育至关重要。尽管这些过程的生理条件相对较为明确，但确切的分子机制仍广泛未知。因此，卵母细胞的生长、分化和成熟是科学争论的主题。大量文献表明，卵母细胞本身在这些过程的机制中起调节作用。因此，本研究通过表达微阵列分析了猪卵母细胞在体外成熟（IVM）过程中的全转录组。选择猪作为实验材料，是因为它在生殖过程方面与人类具有相似性，并且在遗传上与Sus scrofa 也较为接近。从屠宰动物的卵巢中分离卵母细胞，通过 Brilliant Cresyl Blue 试验进行评估，并进行 IVM 处理。一部分卵母细胞被留作“IVM 前”组进行分析。分离卵母细胞 mRNA 并用于微阵列分析，随后通过 RT-qPCR 进行验证。本研究特别关注属于“转录的正调控，DNA 依赖性”、“基因表达的正调控”、“大分子代谢过程的正调控”和“从 RNA 聚合酶 II 启动子转录的正调控”等本体论的基因。FOS、VEGFA、ESR1、AR、CCND2、EGR2、ENDRA、GJA1、INHBA、IHH、INSR、APP、WWTR1、SMARCA1、NFAT5、SMAD4、MAP3K1、EGR1、RORA、ECE1、NR5A1、KIT、IKZF2、MEF2C、SH3D19、MITF 和 PSMB4 均被确定为在这些组中显著改变（倍数变化，>|2|；P<0.05），并且在 IVM 后观察到下调。对表达变化最大的基因进行了分析，并认为它们是与转录调节和大分子代谢过程相关的成熟潜在标记物。
