基于转录组测序筛选母马大小卵泡中颗粒细胞相关上调基因

Transcriptome Sequencing-based Screening of Up-regulated Genes Associated with Large and Small Follicular Granulosa Cells in Horses

在这项研究中,通过采用转录组测序来筛选马的大卵泡和小卵泡的颗粒细胞中的差异表达基因。样本取自三匹蒙古马的大卵泡颗粒细胞和小卵泡颗粒细胞,对上调的基因进行了GO和KEGG信号通路的分析,并使用荧光定量PCR验证了差异表达的基因。结果表明:GO分析分为3大类50组,其中50%的基因参与生物学过程(BP),30%的基因参与细胞组成成分(CC),20%的基因参与分子功能(MF);KEGG信号通路分析发现了9条通路,其中参与细胞外基质受体相互作用通路的基因富集程度最为明显。在小卵泡中筛选出7个上调基因(col1a2、col6a1、col6a2、grem、mgp、bgn、ens)。根据荧光定量PCR的结果,这些基因在小卵泡颗粒细胞中的表达远远高于大卵泡。根据研究结果,在小卵泡中可能需要更丰富的转录物表达,以便为将来转化为优势卵泡的发展可能性提供物质准备。

In this study,transcriptome sequencing was used to screen for differentially expressed genes in granulosa cells of large and small follicles of horses.Samples were taken from the granulosa cells of large and small follicles of three Mongolian horses and the up-regulated genes were analysed for GO and KEGG signalling pathways and the differentially expressed genes were validated using fluorescent quantitative PCR.The results showed that GO analysis was divided into 3 categories and 50 groups,in which 50%genes were involved in biological processes(BP),30%genes were involved in cell components(CC),and 20%genes were involved in molecular functions(MF).The KEGG signalling pathway analysis identified 9 pathways,with the most significant enrichment of genes involved in the extracellular matrix receptor interaction pathway.Seven up-regulated genes(col1a2、col6a1、col6a2、grem、mgp、bgn、ens)were screened in small follicles.According to the results of fluorescence quantitative PCR,the expression of these genes was much higher in the granulosa cells of small follicles than in large follicles.Based on the results of this study,a more abundant expression of transcripts in small follicles may be required in order to prepare ma-terially for the possibility of development of future conversion to dominant follicles.