组织原位杂交和Taq Man实时荧光定量PCR检测enJSRV及其受体HYAL2在妊娠蒙古绵羊绒毛膜中的表达  

Expression of enJSRV and Its Receptor HYAL2 Gene in Chorion of Pregnant Mongolian Sheep by ISH and Real-time PCR

作　　者：齐景伟[1] 徐萌杰[2] 刘淑英[2] 

机构地区：[1]内蒙古农业大学动物科学学院,内蒙古呼和浩特010018 [2]内蒙古农业大学兽医学院,内蒙古呼和浩特010018

出　　处：《动物医学进展》2012年第1期1-6,共6页Progress In Veterinary Medicine

基　　金：国家自然科学基金项目(30960271);国家自然科学基金项目(31160493)

摘　　要：为了探究内源性绵羊肺腺瘤病毒(enJSRV)及其受体HYAL2在妊娠蒙古绵羊胚胎发育过程中的可能作用,本试验应用组织原位杂交技术和Taq Man实时荧光定量PCR对其在不同妊娠时期(30、50、70、90、110、130d)蒙古绵羊绒毛膜的分布定位和表达规律进行了研究。原位杂交结果显示,enJSRV及其受体HYAL2 mRNAs在妊娠30、50、130d蒙古绵羊绒毛膜组织的滋养层巨型双核细胞(BNC)、多核合胞体小半鞘翅(SP)和滋养外胚层(Tr)细胞中均有阳性信号表达;荧光定量PCR结果显示,在妊娠各时期绒毛膜组织中均有enJSRV及受体HYAL2的mRNA表达,通过统计学分析enJSRV mRNA的表达量于妊娠50d时最低,70d和110d相对较高,且差异都极显著(P<0.01),到130d时又降低到起始水平;受体HYAL2的表达于130d时表达量最低,90d相对较高,且差异极显著(P<0.01)。而enJSRV与其受体HYAL2 mRNA表达量之间无线性相关性。研究结果提示,enJSRV在绵羊胚胎发育过程中可能参与调节绒毛膜滋养层细胞的生长并影响胎盘的发育。In order to assess the function of enJSRV and its receptor HYAL2 mRNAs in the chorion of Mongolian ewes throughout gestation.In situ hybridization showed that enJSRV and HYAL2 mRNAs were specifically expressed in the trophoblastic giant binucleated cell,multinucleate syncytial plaque and trophectoderm cell in the chorine on days 30,50,130.By real-time reverse transcription PCR analyses,enJSRV and HYAL2 mRNAs were found to be expressed throughout gestation in the chorion.The abundance of enJSRV mRNA was lower in day 50 and greater in day 70 and 110,and recovered to near original level till day 130.The abundance of receptor HYAL2 mRNA was lower in day 130 and greater in day 90,and the difference is very significant（P〈0.01）.Whereas the level of HYAL2 mRNA was no linear correlation with enJSRV.These results indicated that enJSRV may regulate the growth of trophoblastic cells in the chorion and development of embryos.

关 键 词：内源性绵羊肺腺瘤病毒 透明质酸酶2 绒毛膜 TAQ Man实时荧光定量 原位杂交 

分 类 号：S857.23[农业科学—临床兽医学] Q789[农业科学—兽医学]