牛ODF2和PDF2转录组测序筛选卵泡发育的相关基因

Screening and analysis of genes associated with follicular development in bovine ODF2 and PDF2 transcript

为研究牛卵泡发育过程中影响卵泡发育的重要调控基因及其表达模式,通过B超声波监测并采集牛卵泡发育波中出现偏差期后的第二大卵泡(ODF2)和偏差期前的第二大卵泡(PDF2),建立卵泡颗粒细胞RNA文库并进行高通量测序;测序结果经数据库搜索和差异表达筛选,采用DAVID软件GO和KEGG通路分析,再经Gene Cards功能查询筛选牛卵泡发育的相关基因.结果表明:转录组测序共获得35 325个基因,其中,高表达基因15 536个,进一步筛选出504个差异表达基因;GO分析表明,504个差异表达基因中参与生物过程的基因占39.49%,细胞组分占46.96%,分子功能占13.55%;KEGG通路分析表明,共有97个差异表达基因通过10条信号通路参与牛卵泡发育调控(P<0.05),经Gene Cards进行功能查询后,共筛选出18个与牛卵泡发育密切相关的调控基因.在卵泡发育过程中,PYCARD、MYC、PRICKLE1、TGFBR3、PIK3R3、SOX4、TOPORS、KREMEN1、STK11和NTRK1可能直接参与细胞增殖或凋亡.

The aim of this study was to identify bovine follicular development important genes and their expression patterns. In bovine follicular development wave,the second largest follicle at onset of deviation（ ODF2） and the second largest follicle at onset of predeviation（ PDF2） follicles were collected after B ultrasound monitoring. RNA library of granulesa cells was established and carried out high-throughput sequencing. Sequencing results were searched by database and differentially expressed genes were screened,then analyzed by gene ontology（ GO） and KEGG pathway. Bovine follicular development related genes were screened using GeneCards Function query. Results suggested that 35 325 genes were obtained from the RNA-seq,of which 15 536 significantly expressed genes were selected（ fold change ≥2）,and then 504 differentially expressed genes were selected. GO assignments were used to classify the functions of the 504 genes,and could be categorized into three main catrgories： biological process（ 39.49%）,cellular component（ 49.96%）,molecular function（ 13.55%）. KEGG pathway analysis indicated that 97 differentially expressed genes were involved in bovine follicular development by ten signal paths（ P〈0.05）. Combining with Gene Cards,18 genes have been screened associated with follicle development. PYCARD,MYC,PRICKLE1,TGFBR3,PIK3 R3,SOX4,TOPORS,KREMEN1,STK11,NTRK1 may were directly participated in cell proliferation or apoptosis in the process of follicular development.