Exportin家族鉴定及其在尼罗罗非鱼中的表达分析  

Identification of Exportin Family and Its Expression in Oreochromis niloticus

作　　者：张清清 游慧芹 杜康 陶文静 ZHANG Qingqing;YOU Huiqin;DU Kang;TAO Wenjing(School of Life Sciences,Southwest University/Key Laboratory of Freshwater Fish Reproduction and Development(Ministry of Education),Chongqing 400715,China)

机构地区：[1]西南大学生命科学学院/淡水鱼类资源与生殖发育教育部重点实验室,重庆400715

出　　处：《西南大学学报（自然科学版）》2024年第1期77-86,共10页Journal of Southwest University(Natural Science Edition)

基　　金：国家自然科学基金面上项目(31972778);西南大学大学生创新创业训练计划项目(S202110635298).

摘　　要：输出蛋白(Exportins,XPOs)负责细胞核中蛋白质和RNA的输出,对基因表达调控起着重要的作用,然而Exportin家族的系统进化和表达模式尚不清楚.研究首先对22种代表性动物的Exportin家族进行鉴定并分析了其系统进化,再通过转录组分析和qRT-PCR探究了Exportin家族成员在尼罗罗非鱼各组织中的表达水平,最后通过荧光原位杂交对xpo1a,xpo1b和xpo5在尼罗罗非鱼性腺中的细胞定位进行了分析.结果表明:脊椎动物Exportin家族成员的数量在经过第2轮和第3轮全基因组复制的物种中变化不大,而在经过第4轮时显著增加,暗示Exportin家族成员在第4轮全基因组复制中发生了扩张.xpos主要在精巢中表达,其中xpo1a表达最高;xpos在尼罗罗非鱼孵化后5 d和30 d的表达没有雌雄差异,而在孵化后90,180和300 d出现明显差异,其中负责miRNA出核转运的xpo5在精巢中的表达明显高于卵巢.在尼罗罗非鱼卵巢中,xpo1a,xpo1b和xpo5主要在卵原细胞和卵母细胞中表达;在精巢中,xpo1a和xpo5主要在精原细胞和精母细胞中表达,而xpo1b主要在精细胞中表达.研究系统分析了Exportin家族成员的进化及其在尼罗罗非鱼性腺中的表达,为解析其在脊椎动物生殖发育中的作用奠定了基础.Exportins(XPOs)are responsible for the export of proteins and RNAs in the nucleus,which play an important role in the regulation of gene expression.The evolution and the expression pattern of the XPOs remain unclear.In this study,we firstly characterized XPOs in 22 representative animal species and analyzed its evolution systematically.Then,we analyzed the expression level of XPOs in different tissues of Oreochromis niloticus by transcriptome analysis and qRT-PCR.Finally,we analyzed the cellular location of xpo1a,xpo1b and xpo5 in tilapia gonads by fluorescence in situ hybridization.The results showed that the number of XPOs changed slightly in species with the second and third rounds of whole genome duplication,but increased significantly in species with the fourth round of whole genome duplication,indicating an expansion of this gene family.xpos was mainly expressed in the testis,where the expression of xpo1a was the highest.xpos displayed sexual dimorphic expression at 90,180 and 300 days after hatching(dah).Among them,xpo5,a gene responsible for miRNA export,expressed higher in testis than in ovary.No sexual dimorphic expression of xpos was found in the gonads of tilapia at 5 dah and 30 dah.Our results showed that xpo1a,xpo1b and xpo5 were expressed in oocytes and somatic cells.xpo1a and xpo5 were mainly expressed in spermatogonia and spermatocytes,xpo1b was mainly expressed in spermatocytes.Taken together,in this study,we analyzed the evolution of XPOs and their expressions in tilapia.Our results laid the foundation for elucidating the role of XPOs in the reproduction of vertebrates.

关 键 词：输出蛋白 系统进化分析 表达分析 尼罗罗非鱼 

分 类 号：Q111[生物学—普通生物学]