DPPA2/DPPA4在牛着床前胚胎发育过程中的作用及机制  

Functions of developmental pluripotency-associated 2 and 4(DPPA2/DPPA4)in bovine preimplantation embryo development and their potential mechanisms

作　　者：邱金龙 罗磊 胡冰洁 党燕娜 李爽 史延 王少华 张坤 QIU Jinlong;LUO Lei;HU Bingjie;DANG Yanna;LI Shuang;SHI Yan;WANG Shaohua;ZHANG Kun(Laboratory of Mammalian Molecular Embryology,College of Animal Sciences,Zhejiang University,Hangzhou 310058,Zhejiang,China)

机构地区：[1]浙江大学动物科学学院哺乳动物分子胚胎学实验室,浙江杭州310058

出　　处：《浙江大学学报（农业与生命科学版）》2023年第2期253-260,共8页Journal of Zhejiang University:Agriculture and Life Sciences

基　　金：国家自然科学基金项目(31872348,32072731,32161143032);浙江省自然科学基金重点项目(LZ21C170001)。

摘　　要：多能发育相关因子2和4(developmental pluripotency-associated 2 and 4,DPPA2/DPPA4)是激活类2细胞期胚胎干细胞(2-cell-like embryonic stem cells,2CL ESCs)基因组的重要调控因子,同时调控胚胎干细胞的增殖。DPPA2/DPPA4在牛早期胚胎发育过程中的功能和机制尚不清楚。本研究利用RNA干扰技术结合胚胎显微注射技术敲低牛胚胎中的DPPA2/DPPA4后发现,与阴性对照组相比,敲低组中牛胚胎的8—16细胞阶段卵裂率和囊胚阶段囊胚率无显著差异,但是囊胚总细胞数、滋养层细胞数和内细胞团细胞数均显著减少(P<0.001、P<0.05、P<0.01)。进一步通过RNA测序(RNA-sequencing,RNA-Seq)分析发现,与阴性对照组相比,敲低组的晚期桑葚胚中,NOTCH信号通路的核心转录因子RBPJ(recombination signal binding protein gene for immunoglobulin kappa J region)的表达量显著下调(P<0.05),而本课题组前期实验证明,RBPJ敲低会导致牛早期胚胎的囊胚质量严重受损。本研究结果揭示了DPPA2/DPPA4在牛着床前胚胎发育中可能通过调节RBPJ的表达来影响细胞增殖。Developmental pluripotency-associated 2 and 4(DPPA2/DPPA4)are important regulators of genome activation in 2-cell-like embryonic stem cells(2CL ESCs),meanwhile,they can regulate the proliferation of ESCs.However,the functions of DPPA2/DPPA4 during early bovine embryo development and their potential mechanisms remain unclear.In this study,RNA interference technology combined with embryo microinjection technology were used to knock down DPPA2/DPPA4 in bovine embryos.It was found that,compared with the negative control(NC)group,there was no significant difference in the cleavage rate from 8-cell to 16-cell stages and blastocyst rate at the blastocyst stage,but the total cell number,trophoblast cell number,and inner cell mass(ICM)cell number of blastocysts were significantly decreased in the knockdown(KD)group(P<0.001,P<0.05,and P<0.01).RNA-sequencing analysis further showed that,when compared with the NC group,the core transcription factor RBPJ(recombination signal binding protein gene for immunoglobulin kappa J region)of NOTCH signaling pathway was significantly down-regulated(P<0.05)in the late morula stage at the KD group.The previous studies demonstrated that RBPJ knockdown would lead to severe damage to blastocyst quality of early bovine embryos.Therefore,the results revealed that DPPA2/DPPA4 can affect cell proliferation by regulating the expression of RBPJ during bovine preimplantation embryo development.

关 键 词：牛胚胎 着床前 多能发育相关因子2和4 RBPJ基因 

分 类 号：S823.91[农业科学—畜牧学]