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“生物黏附”是输卵管上皮细胞(OECs)长期原代体外培养过程中受到严格调控的分子过程:一项转录组学和蛋白质组学研究。

"Biological Adhesion" is a Significantly Regulated Molecular Process during Long-Term Primary In Vitro Culture of Oviductal Epithelial Cells (Oecs): A Transcriptomic and Proteomic Study.

机构信息

Department of Histology and Embryology, Poznan University of Medical Sciences, 60-781 Poznan, Poland.

Department of Inorganic and Analytical Chemistry, Poznan University of Medical Sciences, 60-870 Poznań, Poland.

出版信息

Int J Mol Sci. 2019 Jul 10;20(14):3387. doi: 10.3390/ijms20143387.

DOI:10.3390/ijms20143387
PMID:31295879
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6678391/
Abstract

Oviductal epithelial cells (OECs) actively produce stimulating and protecting factors, favoring survival and viability of gametes and early embryos. The oviduct participates in the initial reproductive events, which strongly depends on adhesion. The analysis of differential gene expression in OECs, during long-term in vitro culture, enables recognition of new molecular markers regulating several processes, including "biological adhesion". Porcine oviducts were stained with hematoxylin and eosin, as well as with antibodies against epithelial markers. Then, OECs were long-term in vitro cultured and after 24 h, 7, 15, and 30 days of culture were subjected to transcriptomic and proteomic assays. Microarrays were employed to evaluate gene expression, with Matrix-assisted laser desorption/ionization-time of light (MALDI-TOF) mass spectrometry applied to determine the proteome. The results revealed proper morphology of the oviducts and typical epithelial structure of OECs during the culture. From the set of differentially expressed genes (DEGs), we have selected the 130 that encoded proteins detected by MALDI-TOF MS analysis. From this gene pool, 18 significantly enriched gene ontology biological processes (GO BP) terms were extracted. Among them we focused on genes belonging to "biological adhesion" GO BP. It is suggested that increased expression of studied genes can be attributed to the process of intensive secretion of substances that exhibit favorable influence on oviductal environment, which prime gametes adhesion and viability, fertilization, and early embryo journey.

摘要

输卵管上皮细胞 (OECs) 会积极产生刺激和保护因子,有利于配子和早期胚胎的存活和活力。输卵管参与初始生殖事件,这强烈依赖于黏附。在长期体外培养过程中分析 OECs 中的差异基因表达,可识别出调节包括“生物黏附”在内的多个过程的新分子标记。对猪输卵管进行了苏木精和伊红染色,以及上皮标记物抗体染色。然后,对 OECs 进行长期体外培养,培养 24 h、7 d、15 d 和 30 d 后进行转录组和蛋白质组学检测。采用微阵列评估基因表达,采用基质辅助激光解吸/电离-飞行时间 (MALDI-TOF) 质谱法确定蛋白质组。结果显示,在培养过程中,输卵管具有适当的形态和典型的 OECs 上皮结构。从差异表达基因 (DEGs) 中,我们选择了 130 个由 MALDI-TOF MS 分析检测到的编码蛋白的基因。从这个基因池中提取了 18 个显著富集的基因本体生物学过程 (GO BP) 术语。其中,我们重点关注属于“生物黏附”GO BP 的基因。研究表明,所研究基因的表达增加可能归因于物质的强烈分泌过程,这些物质对输卵管环境具有有利影响,促进配子的黏附和活力、受精和早期胚胎发育。

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