Huang Gao-Bo, Quan Li, Zeng Yong-Lian, Yang Jian, Lu Ke-Huan, Lu Sheng-Sheng
State Key Laboratory for Conservation and Utilisation of Subtropical Agro-Bioresources, Guangxi University, Nanning, Guangxi 53004, China.
Tiandiyang Biotechnology Co. Ltd, Nanning, Guangxi 530004, China.
Reprod Fertil Dev. 2016 Mar;28(3):302-9. doi: 10.1071/RD14051.
During reprogramming, there is exchange of histone H1c and the oocyte-specific linker histone, and H1c may play a critically important role in the reprogramming process of somatic cell nuclear transfer (SCNT). The aim of the present study was to investigate the role of the H1c gene in SCNT reprogramming in Chinese swamp buffalo (Bubalus bubalis) using RNA interference (RNAi). Chinese swamp buffalo H1c gene sequences were obtained and H1c-RNAi vectors were designed, synthesised and then transfected into a buffalo fetal skin fibroblast cell line. Expression of H1c was determined by real-time polymerase chain reaction to examine the efficiency of vector interference. These cells were then used as a nuclear donor for SCNT so as to observe the further development of SCNT embryos. Inhibition of H1c gene expression in donor cells significantly improved the developmental speed of embryos from the 1-cell to 8-cell stage. Furthermore, compared with the control group, inhibition of H1c gene expression significantly reduced the blastocyst formation rate. It is concluded that linker histone H1c is very important in SCNT reprogramming in Chinese swamp buffalo. Correct expression of the H1c gene plays a significant role in preimplantation embryonic development in B. bubalis.
在重编程过程中，组蛋白H1c与卵母细胞特异性连接组蛋白发生交换，并且H1c可能在体细胞核移植（SCNT）的重编程过程中发挥至关重要的作用。本研究的目的是利用RNA干扰（RNAi）研究H1c基因在中国水牛（Bubalus bubalis）SCNT重编程中的作用。获得了中国水牛H1c基因序列，设计、合成了H1c-RNAi载体，然后将其转染到水牛胎儿皮肤成纤维细胞系中。通过实时聚合酶链反应测定H1c的表达，以检测载体干扰效率。然后将这些细胞用作SCNT的核供体，以观察SCNT胚胎的进一步发育。供体细胞中H1c基因表达的抑制显著提高了胚胎从1细胞到8细胞阶段的发育速度。此外，与对照组相比，H1c基因表达的抑制显著降低了囊胚形成率。得出的结论是，连接组蛋白H1c在中国水牛的SCNT重编程中非常重要。H1c基因的正确表达在水牛植入前胚胎发育中起重要作用。
