体外抑制Piwi基因对鸡原始生殖细胞相关基因表达的影响  

In Vitro Inhibition of Piwi Gene Expression and the Influence on Related Genes in Primordial Germ Cells

作　　者：李建超[1] 张颖[1] 戴爱琴[1] 王洪志[1] 翟飞[1] 华登科[1] 夏明秀[1] 常国斌[1] 陈国宏[1] 

机构地区：[1]扬州大学动物科学与技术学院,扬州225009

出　　处：《畜牧兽医学报》2014年第6期871-878,共8页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(31372297;31301966);国家科技支撑计划(2011BAD28B03)

摘　　要：本研究以鸡原始生殖细胞(Primordial germ cells,PGCs)为材料进行Piwi基因的干扰,并探究Piwi和生殖特异性标记基因CVH、Dazl和CDH及调控干细胞多能性转录因子基因Sox2、Nanog和PouV间的关系。由孵化至19期的鸡胚分离出PGCs,经鉴定后,利用RNA干扰技术抑制Piwi基因表达,相对实时荧光定量PCR检测目的基因的表达。结果发现,体外抑制Piwi后,CVH、Dazl和Nanog表达量显著增加(P<0.05),PouV和Sox2表达量显著下降(P<0.05),CDH表达量变化不显著(P>0.05)。本研究在家禽PGCs中建立Piwi基因干扰模型,并发现其干扰后影响生殖特异性标记基因和干性转录因子的表达,为其在生殖生物学中的功能研究提供了新的依据。This study was designed to interfere Piwi gene in chicken primordial germ cells(PGCs)and explore its interaction with CVH,Dazl,CDH,Sox2,Nanogand PouVgenes.PGCs was isolated from 19 stage chicken embryo,and after identification of PGCs,we analyzed expression of CVH,Dazl,CDH,Nanog,Sox2 and PouVunder the condition of Piwi interference in PGCs by using real time quantitative PCR.The results showed that expression of CVH,Dazl and Nanoggenes increased significantly(P<0.05),while the expression of PouVand Sox2 genes decreased significantly followed with Piwi interference in PGCs(P<0.05).Collectively,we constructed a Piwi gene interference model in vitro and found,and which influenced expression of CVH,Dazl,CDH,Sox2,Nanogand PouV,which provided new basis for studying Piwi gene function in reproductive biology.

关 键 词：鸡 原始生殖细胞 PIWI RNA干扰 

分 类 号：S831[农业科学—畜牧学]