Department of Veterinary Surgery, Institute of Veterinary Medicine, Nicolaus Copernicus University in Torun, Torun, Poland.
Department of Anatomy, Poznan University of Medical Sciences, Poznan, Poland.
Theriogenology. 2020 May;148:76-88. doi: 10.1016/j.theriogenology.2020.02.044. Epub 2020 Mar 3.
The oogenesis and folliculogenesis are closely linked and occur simultaneously in the growing ovarian follicles. Biochemical and morphological changes in oocytes (OC) and surrounding granulosa cells (GCs) are highly complex and depend on many factors, including intercellular communication. GCs are cells with many functions, often crucial for the proper viability of the oocyte and subsequent positive fertilization. The purpose of this study was to analyze gene expression in porcine GCs, to define differentially expressed genes belongs to the "cell growth", "aging", "positive regulation of cell death", "apoptotic process", "regulation of cell death", "cell death" and "negative regulation of cell death" ontology groups during the short - term primary in vitro culture. Microarrays were employed to study the transcriptome contained in the total RNA of the cultured GCs. Ovaries were obtained after slaughter, from 40 gilts of swine aged 170 days. The cells were obtained through puncture of the ovaries, collection of follicular fluid, removal of the cumulus - oocyte complexes and centrifugation. The cells were then cultured in vitro. The RNA material was obtained before the culture was established (0h) and then after 48h, 96h and 144h of its course. From 182 differently expressed genes belonging to the these ontology groups, we have selected POSTN, FN1, FMOD, ITGB3, DCN, SERPINB2, SFRP2, IGFBP5, EMP1, and CCL2 which were upregulated, as well as DAPL1, ESR1, IHH, TGFBR3, PPARD, PDK4, TXNIP, IFIT3, CSRNP3, and TNFSF10 genes whose expression was downregulated during the time of in vitro culture of the GCs. The significance of the differential gene expression is to provide new information on the molecular aspects of in vitro granulosa cell culture.
卵母细胞发生和卵泡发生密切相关,并同时发生在生长中的卵巢卵泡中。卵母细胞(OC)和周围颗粒细胞(GC)的生化和形态变化非常复杂,取决于许多因素,包括细胞间通讯。GC 是具有多种功能的细胞,通常对卵母细胞的适当存活和随后的受精阳性至关重要。本研究旨在分析猪 GC 的基因表达,定义在短期原代体外培养过程中属于“细胞生长”、“衰老”、“细胞死亡的正调控”、“凋亡过程”、“细胞死亡的调控”、“细胞死亡”和“细胞死亡的负调控”本体论组的差异表达基因。使用微阵列研究培养 GC 总 RNA 中的转录组。卵巢取自 170 天大的 40 头母猪屠宰后,通过穿刺卵巢、收集卵泡液、去除卵丘-卵母细胞复合体和离心获得细胞。然后将细胞在体外培养。在建立培养物之前(0 小时)获得 RNA 材料,然后在其过程中 48 小时、96 小时和 144 小时后获得。从属于这些本体论组的 182 个差异表达基因中,我们选择了 POSTN、FN1、FMOD、ITGB3、DCN、SERPINB2、SFRP2、IGFBP5、EMP1 和 CCL2,这些基因上调,以及 DAPL1、ESR1、IHH、TGFBR3、PPARD、PDK4、TXNIP、IFIT3、CSRNP3 和 TNFSF10 基因在 GC 的体外培养过程中表达下调。差异基因表达的意义在于为体外颗粒细胞培养的分子方面提供新的信息。