利用数字基因表达谱对鸡卵泡发育相关基因的筛选

摘要

To identify the key genes involved in follicular recruitment,hierarchies and ovulation, the differently expressed genes profile in pre-hierarchical follicles(small white follicles,SWF)and the pre-ovulatory follicles(the first largest follicle,F1)were obtained by using Solexa/Illusion RNA-seqencing.In this study,784 up-regulated and 1 526 down-regulated genes in F1 follicles were identified compared to SWF follicles.The genes up-regulated more than five folds included PTGS1 ,TEK,VEGF-A ,MMP-3 and CDK1 .As the same down-regulated genes included β-catenin,ZO-1 ,Nectin-3 ,PTPX15 and Gadd45 .GO analysis indicated that the protein encoded by the differently expressed genes mainly involved in biological processes including activation of different transferases,molecular connectivity enzymatic hydrolysis and proteolysis.The pathway analysis suggested that the differently expressed genes mainly took part in cell adherence junction,p53 pathway,protein synthesis,and cell cycle pathways during follicle development. Furthermore,the expression patterns of MMP-3 and MMP-9 genes in chicken hierarchy follicles showed that the two genes expression levels increased during follicular maturation,MMP-3 expression level reached maximum in the first largest follicle(F1),while MMP-9 expression level continued to rise in POF1 and POF2 after ovulation.%为筛选禽类卵泡募集、等级化维持及排卵过程中的关键基因，通过数字基因表达谱检测鸡卵巢内等级前小白卵泡（SWF）和发育到最大期的等级卵泡（F1）中的差异表达基因。结果显示，随着卵泡的成熟，显著上调表达的基因有784个，显著下调的基因有1526个。基因表达差异倍数达到5倍以上的上调基因包括 PTGS1、TEK、VEGF A 、MMP 3、CDK1等；下调基因包括β cate-nin 、ZO 1、Nectin 3、PTPX15、Gadd45等。对差异表达基因进行基因本体（GO）生物学分析和信号通路显著富集分析发现，差异表达的基因所编码的蛋白主要参与生物学催化反应、分子连接、转移酶活性、水解等生物学过程；主要与细胞黏合通路、p53信号通路、蛋白合成通路及细胞周期通路等信号通路有关。对卵泡发育过程中的基质金属蛋白酶家族（MMPs）的 MMP 3与 MMP 9基因表达变化进行测定发现，MMP 3基因表达量在卵泡发育过程中显著升高，排卵后的卵泡中显著下降；MMP 9基因表达量则持续显著升高，在排卵后第2天的卵泡中下降，但下降不明显。