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调控激素刺激和蛋白信号反应的基因在猪卵母细胞体外成熟过程中表现出不同的表达模式,这一结果得到了脂质浓度的证实。

Genes regulating hormone stimulus and response to protein signaling revealed differential expression pattern during porcine oocyte in vitro maturation, confirmed by lipid concentration.

机构信息

Division of Infertility and Reproductive Endocrinology, Department of Gynecology, Obstetrics and Gynecological Oncology, Poznan University of Medical Sciences, Poznan, Poland.

Department of Obstetrics and Gynecology, University Hospital and Masaryk University, Brno, Czech Republic.

出版信息

Histochem Cell Biol. 2020 Jul;154(1):77-95. doi: 10.1007/s00418-020-01866-w. Epub 2020 Mar 18.

Abstract

Genes influencing oocyte maturation may be valuable for predicting their developmental potential, as well as discerning the mechanistic pathways regulating oocyte development. In the presented research microarray gene expression analysis of immature and in vitro matured porcine oocytes was performed. Two groups of oocytes were compared in the study: before (3 × n = 50) and after in vitro maturation (3 × n = 50). The selection of viable oocytes was performed using the brilliant cresyl blue (BCB) test. Furthermore, microarrays and RT-qPCR was used to analyze the transcriptome of the oocytes before and after IVM. The study focused on the genes undergoing differential expression in two gene-ontology groups: "Cellular response to hormone stimulus" and "Cellular response to unfolded protein", which contain genes that may directly or indirectly be involved in signal transduction during oocyte maturation. Examination of all the genes of interest showed a lower level of their expression after IVM. From the total number of genes in these gene ontologies ten of the highest change in expression were identified: FOS, ID2, BTG2, CYR61, ESR1, AR, TACR3, CCND2, EGR2 and TGFBR3. The successful maturation of the oocytes was additionally confirmed with the use of lipid droplet assay. The genes were briefly described and related to the literature sources, to investigate their potential roles in the process of oocyte maturation. The results of the study may serve as a basic molecular reference for further research aimed at improving the methods of oocyte in vitro maturation, which plays an important role in the procedures of assisted reproduction.

摘要

影响卵母细胞成熟的基因可用于预测其发育潜能,以及辨别调节卵母细胞发育的机制途径。本研究对未成熟和体外成熟的猪卵母细胞进行了基因表达谱分析。在研究中比较了两组卵母细胞:体外成熟前(3×n=50)和体外成熟后(3×n=50)。使用亮甲酚蓝(BCB)测试选择有活力的卵母细胞。此外,还使用微阵列和 RT-qPCR 分析了 IVM 前后卵母细胞的转录组。该研究集中在两个基因本体组中差异表达的基因上:“细胞对激素刺激的反应”和“细胞对未折叠蛋白的反应”,其中包含可能直接或间接参与卵母细胞成熟过程信号转导的基因。对所有感兴趣基因的检查表明,IVM 后其表达水平降低。在这些基因本体中,从总基因数中确定了表达变化最大的前 10 个基因:FOS、ID2、BTG2、CYR61、ESR1、AR、TACR3、CCND2、EGR2 和 TGFBR3。还使用脂滴测定法进一步确认了卵母细胞的成熟。简要描述了这些基因,并与文献来源相关联,以研究它们在卵母细胞成熟过程中的潜在作用。该研究结果可为进一步研究改善卵母细胞体外成熟方法提供基础分子参考,因为这在辅助生殖程序中起着重要作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8866/7343741/7c443e1a3621/418_2020_1866_Fig1_HTML.jpg

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