基于转录组测序筛选山羊卵泡发育相关上调基因

Transcriptome Sequencing Screened the Up-Regulated Genes Related to Follicle Development in Goats

本研究旨在通过转录组测序筛选并研究与卵泡发育相关的上调表达基因。以3只健康的1岁龄贵州白山羊为供试对象,在其发情3 d后,屠宰并采集卵巢中的优势卵泡与从属卵泡,分离颗粒细胞,转录组测序。通过生物信息学分析,并对其中的上调表达基因进行GO、KEGG信号通路分析,经实时荧光定量PCR对筛选出的上调表达基因进行验证。结果显示:GO分析共分为三大类别44组,其中参与生物学过程(BP)的基因占72.7%,参与细胞组分(CC)的基因占13.6%,参与分子功能(MF)的基因占13.6;KEGG信号通路分析发现20条通路,其中参与核糖体通路基因富集最为显著。共筛选出4个可能促进山羊卵泡发育的基因(CALM2、TUT7、LUM和PDLIM3)。荧光定量PCR结果表明,TUT7和CALM2在优势卵泡颗粒细胞中的表达量显著高于从属卵泡。本研究推测获得的2个上调表达基因可能会促进卵泡的发育,从而导致优势卵泡被选择。

To screen and study the up-regulated genes associated with follicular development using transcriptome sequencing. Three healthy 1-year old guizhou white goat was selected as the test subjects. Estrus after 3 days, the goats were slaughtered, and granulosa cells were isolated from dominant and subordinate follicles in every pair of ovaries,and then transcriptome sequencing was performed. After bioinformatics analysized, up-regulated genes were analyzed by GO and KEGG signaling pathways, and verified by RT-PCR. GO analysis was divided into three categories with 44 groups: genes involved in biological process(BP) accounted for 72.7%, genes involved in cell component(CC) accounted for 13.6%, and genes involved in molecular function(MF) accounted for 13.6%. KEGG signaling pathway analysis found 20 pathways, among which gene enrichment in ribosome pathway was the most significant. Four genes that may promote follicle development were selected from up-regulated expression genes, namely CALM2, TUT7, LUM and PDLIM3. RT-PCR Results showed that TUT7 and CALM2 were significantly higher in DF granulosa cells than in SF(P<0.05).We speculated that the two up-regulated genes obtained may promote the follicle development in goat, and ultimately lead to the selection of dominant follicles.