Division of Infertility and Reproductive Endocrinology, Department of Gynecology, Obstetrics and Gynecological Oncology, Poznan University of Medical Sciences, 60-535 Poznań, Poland.
Department of Obstetrics and Gynecology, University Hospital and Masaryk University, 601 77 Brno, Czech Republic.
Int J Mol Sci. 2019 May 7;20(9):2238. doi: 10.3390/ijms20092238.
This paper aims to identify and describe new genetic markers involved in the processes of protein expression and modification reflected in the change of mitochondrial activity before and after in vitro maturation of the oocyte. Porcine oocytes collected from the ovaries of slaughtered landrace gilts were subjected to the process of in vitro maturation. Transcriptomic changes in the expression profile of oocyte genes involved in response to hypoxia, the transmembrane protein receptor serine threonine kinase signaling pathway, the "transforming growth factor β receptor signaling pathway", "response to protein stimulus", and "response to organic substance" were investigated using microarrays. The expression values of these genes in oocytes was analyzed before (immature) and after (mature) in vitro maturation, with significant differences found. All the significantly altered genes showed downregulation after the maturation process. The most changed genes from these gene ontologies, , , , , , , , , , , were chosen to be further validated, described and related to the literature. Additionally, the mitochondrial activity of the analyzed oocytes was measured using specific dyes. We found that the mitochondrial activity was higher before the maturation process. The analysis of these results and the available literature provides a novel insight on the processes that occur during in vitro oocyte maturation. While this knowledge may prove to be useful in further research of the procedures commonly associated with in vitro fertilization procedures, it serves mostly as a basic reference for further proteomic, in vivo, and clinical studies that are necessary to translate it into practical applications.
本文旨在鉴定和描述新的遗传标记物，这些标记物涉及蛋白质表达和修饰过程，反映了卵母细胞体外成熟前后线粒体活性的变化。从屠宰长白母猪卵巢中采集猪卵母细胞，进行体外成熟过程。使用微阵列技术研究卵母细胞中参与缺氧反应、跨膜蛋白受体丝氨酸苏氨酸激酶信号通路、“转化生长因子 β 受体信号通路”、“对蛋白质刺激的反应”和“对有机物质的反应”的基因表达谱的转录组变化。分析这些基因在卵母细胞体外成熟前后（不成熟和成熟）的表达值，发现存在显著差异。所有成熟后表达显著改变的基因均表现出下调。从这些基因本体中选择变化最大的基因进行进一步验证、描述和文献相关分析，,,,,,,,,,, 。此外，使用特定染料测量分析卵母细胞的线粒体活性。我们发现成熟前的线粒体活性更高。分析这些结果和现有文献为体外卵母细胞成熟过程中发生的过程提供了新的见解。虽然这些知识可能对进一步研究与体外受精程序相关的程序有用，但它主要是进一步进行蛋白质组学、体内和临床研究的基础参考，以便将其转化为实际应用。
