牛Sry亚细胞定位及其对Sox9基因表达的影响  

Subcellur location of bovine Sry and its regulation on Sox9 expression

作　　者：韩凤桐[1] 吴宁[1] 廖冰[1] 林秀坤[1] 刘娣 

机构地区：[1]中国农业科学院北京畜牧兽医研究所 [2]黑龙江省农业科学院

出　　处：《动物学报》2008年第5期890-896,共7页ACTA ZOOLOGICA SINICA

基　　金：国家自然科学基金资助项目(No.30671501)~~

摘　　要：为鉴别牛Sry基因的下游目的基因,并确定Sry蛋白在细胞内的定位区域,本试验构建pcDNA3.1-Sry表达载体和pEGFP-N1-Sry亚细胞定位表达载体,在原代培养的生殖嵴细胞和卵巢颗粒细胞中进行Sry过表达,并对性别控制相关基因在Sry过表达前后的表达水平变化进行检测,包括Sf1(Steroidogenic fator-1)、Gata4(GATA binding protein 4)、Wt1(Wilms tumor gene)、Sox9(Sry-related HMG box-9)、Amh(Anti Mullerian Hormone)和Dax1(Dosage sensitive sex reversal locus-1);并在两种细胞中进行了Sry亚细胞定位研究。Sry在两种细胞中过表达后,在生殖嵴细胞中Sox9的表达水平被显著上调,但在卵巢颗粒细胞中未观察到Sox9基因被上调,而其他几个基因的表达水平未受Sry影响;亚细胞定位结果表明牛Sry主要分布在细胞核内。本试验说明牛Sry可能间接调控Sox9表达,或存在其他不需要Sry参与的调控通路调控Sox9表达;牛Sry蛋白起转录因子作用。This study attempted to identify the putative targets of Sry in the process of bovine sex-determination and to analyze the localization of Sty protein in cells. The Sry gene was overexpressed by transfecting pcDNA3.1-Sty in primary cultured genital ridge cells and granulosa cells, and six genes which were identified as important in sex determination in mammals were analyzed, including 5f1 （Steroidogenicfator-1）, Gata4 （ GATA binding protein 4）, Wt1 （ Wilms＇ tumor gene ）, Sox9 （ Sry-related HMG box-9）, Amh （Anti Mullerian Hormone ）, and Daxl （Dosage sensitive sex reversal locus-1 ）. Subcellular localization of the expressed Sty protein in genital ridge cells and granulosa cells was also analyzed. When Sry was overexpressed, significant increases of Sox9 mRNA expression were observed in genital ridge cells, but not in granulosa cells. Other gene expressions checked were not affected by Sry overexpression. Subcellular localization analyses indicated that Sty protein was mainly detected in the nuclei of both genital ridge cells and granulosa ceils. Our results indicated that Sry might regulate Sox9 indirectly, or there might be another regulatoty pathway for Sox9 expression without the involvement of Sry. Sty probably works as a transcription factor in bovine sex determination

关 键 词：牛 SRY 过表达 亚细胞定位 SOX9 

分 类 号：Q344[生物学—遗传学]