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编码负责猪卵巢颗粒细胞结构的蛋白质的新基因标记的转录组图谱。

Transcriptomic Profile of New Gene Markers Encoding Proteins Responsible for Structure of Porcine Ovarian Granulosa Cells.

作者信息

Kulus Jakub, Kulus Magdalena, Kranc Wiesława, Jopek Karol, Zdun Maciej, Józkowiak Małgorzata, Jaśkowski Jędrzej M, Piotrowska-Kempisty Hanna, Bukowska Dorota, Antosik Paweł, Mozdziak Paul, Kempisty Bartosz

机构信息

Department of Diagnostics and Clinical Sciences, Institute of Veterinary Medicine, Nicolaus Copernicus University in Torun, 87-100 Torun, Poland.

Department of Veterinary Surgery, Institute of Veterinary Medicine, Nicolaus Copernicus University in Torun, 87-100 Torun, Poland.

出版信息

Biology (Basel). 2021 Nov 20;10(11):1214. doi: 10.3390/biology10111214.

Abstract

The extracellular matrix (ECM) in granulosa cells is functionally very important, and it is involved in many processes related to ovarian follicle growth and ovulation. The aim of this study was to describe the expression profile of genes within granulosa cells that are associated with extracellular matrix formation, intercellular signaling, and cell-cell fusion. The material for this study was ovaries of sexually mature pigs obtained from a commercial slaughterhouse. Laboratory-derived granulosa cells (GCs) from ovarian follicles were cultured in a primary in vitro culture model. The extracted genetic material (0, 48, 96, and 144 h) were subjected to microarray expression analysis. Among 81 genes, 66 showed increased expression and only 15 showed decreased expression were assigned to 7 gene ontology groups "extracellular matrix binding", "extracellular matrix structural constituent", "binding, bridging", "cadherin binding", "cell adhesion molecule binding", "collagen binding" and "cadherin binding involved in cell-cell adhesion". The 10 genes with the highest expression (POSTN, ITGA2, FN1, LAMB1, ITGB3, CHI3L1, PCOLCE2, CAV1, DCN, COL14A1) and 10 of the most down-regulated (SPP1, IRS1, CNTLN, TMPO, PAICS, ANK2, ADAM23, ABI3BP, DNAJB1, IGF1) were selected for further analysis. The results were validated by RT-qPCR. The current results may serve as preliminary data for further analyses using in vitro granulosa cell cultures in assisted reproduction technologies, studies of pathological processes in the ovary as well as in the use of the stemness potential of GCs.

摘要

颗粒细胞中的细胞外基质(ECM)在功能上非常重要,并且参与许多与卵泡生长和排卵相关的过程。本研究的目的是描述颗粒细胞内与细胞外基质形成、细胞间信号传导和细胞-细胞融合相关的基因表达谱。本研究的材料是从商业屠宰场获得的性成熟猪的卵巢。从卵巢卵泡中分离出的颗粒细胞(GCs)在原代体外培养模型中进行培养。提取的遗传物质(0、48、96和144小时)进行微阵列表达分析。在81个基因中,66个基因表达增加,只有15个基因表达减少,这些基因被归为7个基因本体组,即“细胞外基质结合”、“细胞外基质结构成分”、“结合、桥接”、“钙黏蛋白结合”、“细胞黏附分子结合”、“胶原蛋白结合”和“参与细胞-细胞黏附的钙黏蛋白结合”。选择表达最高的10个基因(POSTN、ITGA2、FN1、LAMB1、ITGB3、CHI3L1、PCOLCE2、CAV1、DCN、COL14A1)和下调最明显的10个基因(SPP1、IRS1、CNTLN、TMPO、PAICS、ANK2、ADAM23、ABI3BP、DNAJB1、IGF1)进行进一步分析。结果通过RT-qPCR进行验证。目前的结果可作为辅助生殖技术中使用体外颗粒细胞培养进行进一步分析、卵巢病理过程研究以及颗粒细胞干性潜能利用的初步数据。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b1b6/8615192/c1d5f0baf4a9/biology-10-01214-g0A1.jpg

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