Scriptaid处理牦牛成纤维细胞对组蛋白乙酰化及重编程的影响  

Effects of Scriptaid on Histone Acetylation and Reprogramming of Yak(Bos grunniens) Fibroblast Cells

作　　者：高川[1] 熊显荣[1] 符梅[1] 王树茂[1] 李宇[1] 李键[1] 

机构地区：[1]西南民族大学生命科学学院,动物遗传育种重点实验室,成都610041

出　　处：《农业生物技术学报》2013年第1期62-68,共7页Journal of Agricultural Biotechnology

基　　金：国家科技支撑计划(No.2012BAD13B06);西南民族大学中央高校基本科研业务费专项(No.82000860和No.11NZYQN33)

摘　　要：供体细胞的不完全重编程是克隆动物成功率低的主要原因。本研究旨在探讨组蛋白去乙酰化酶抑制剂Scriptaid对牦牛(Bos grunniens)成纤维细胞重编程的影响,以及Scriptaid处理供体细胞对牦牛-黄牛(Bos taurus)异种核移植(interspecies somatic cell nuclear transfer,iSCNT)胚胎体外发育的影响。分别利用0、500、1000、1500和3000nmol/LScriptaid处理牦牛胎儿成纤维细胞,观察细胞形态、统计细胞活率、测定细胞周期、检测组蛋白乙酰化水平,并以处理后的成纤维细胞进行体细胞核移植,检测重构胚体外发育水平。结果显示,经Scriptaid处理24h后,细胞形态发生了改变,细胞增殖受到明显抑制,大量细胞被阻滞在G0/G1期;随着Scriptaid处理浓度的增加,细胞组蛋白H3K9ac乙酰化水平逐渐提高;以1000nmol/L Scriptaid处理的成纤维细胞作为供体细胞后的囊胚率显著高于其他组(P<0.05)。研究结果表明,Scriptaid能显著提高牦牛成纤维细胞的乙酰化水平,以1000nmol/L Scriptaid处理供体细胞显著提高了牦牛-黄牛异种克隆胚胎的体外发育。本研究初步证实Scriptaid是通过提高成纤维细胞的乙酰化水平来促进核的重编程,为研究供体细胞的重编程以及异种克隆牦牛提供理论依据。It has been widely suggested that the incompletely reprogramming of donor cell was the major reason of the inefficiency of nuclear transfer. The present study was to investigate the effect of histone deacetylase inhibitor （Scriptaid） on the reprogramming of yak （Bos grunniens） fetal fibroblast cells, and the in vitro developmental capability of yak-bovine （Bos grunniens） interspecies somatic cell nuclear transfer（iSCNT） embryos after donor cells were treated with Scriptaid. The yak fetal fibroblasts were treated with 0, 500, 1 000, 1 500 and 3 000 nmol/L Scriptaid for 24 h, respectively. Then observed cell morphology, accounted the survival rate, analyzed cell cycle by flow cytometry, detected acetylation level of H3K9ac by immunofluorescence, and evaluated the developmental competence of iSCNT embryos, which showed that the cell morphology was changed significantly, the cell became flatten and the area of nuclear was larger after Scriptaid treatment; cell proliferation was inhibited gradually and most cells were arrested at G0/G1 stage after Scriptaid treatment for 24 h; the acetylation level of H3K9ac was enhanced along with the concentrations of Scriptaid; the cleavage and blastocyst formation rate of iSCNT embryos improved significantly when donor cells treated with 1 000 nmol/L Scriptaid for 24 h （P〈0.05）. The results suggested that Scriptaid can significantly improve the histone acetylation level of yak fetal fibroblast cells, pre-treated donor cells with 1 000 nmol/L Scriptaid was helpful for improving in vitro development of yak-bovine iSCNT embryos. It was concluded that Scriptaid enhances the reprogramming in terms of high histone acetylation of donor cells, and it is useful for studying reprogramming and a novel way to improve the efficiency of yak iSCNT.

关 键 词：Scriptaid 组蛋白乙酰化 重编程 异种体细胞核移植 

分 类 号：Q785[生物学—分子生物学]