畜牧与饲料科学 ›› 2017, Vol. 38 ›› Issue (5): 15-15.doi: 10.12160/j.issn.1672-5190.2017.05.004

• 基础科学 • 上一篇    下一篇

Scriptaid对猪移植体细胞核在胚胎外发育的影响

卢富春;史兆国   

  1. 甘肃农业大学动物科学技术学院,甘肃兰州730070
  • 出版日期:2017-05-20 发布日期:2017-05-20
  • 通讯作者: 卢富春
  • 作者简介:卢富春(1987-),男,硕士研究生,主要研究方向为畜禽养殖。 通讯作者:史兆国(1966-),男,教授,硕士,硕士生导师,主要研究方向为家禽营养与生产。

Effect of Scriptaid on Extraembryonic Development of Transplanted Somatic Cell Nuclear in Swine

LU Fu-chun, SHI Zhao-guo (College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, China)   

  • Online:2017-05-20 Published:2017-05-20

摘要: 组蛋白去乙酰化酶(histone deacetylases,HDACs)是调控基因的关键蛋白酶,乙酰化是一种可逆的蛋白共价修饰。组蛋白去乙酰化酶可以改变特定基因的转录和表达水平,诱导细胞的分化和凋亡。为了检测一种低毒新型的组蛋白去乙酰化酶抑制剂Scriptaid对克隆猪胚胎发育的影响,进行了不同浓度和不同时间的处理,并在体外检测了胚胎分裂率和囊胚发育能力。研究以对照试验作为基础,以凌源禾丰种猪场长白猪胎儿成纤维作为供体细胞,然后通过体细胞核移植(SCNT)技术获得体外发育胚胎。将重构胚胎以不同浓度(0、200、500、700、900 nmol/L)Scriptaid处理后,并进行不同时间(0、15、36、72 h)的培养,然后通过观察不同处理浓度和处理时间下胚胎在2细胞阶段分裂率和囊胚发育率。再通过Real-time PCR检测2细胞阶段未处理组与Scriptaid处理组的Oct4、Sox2两个因子的相对表达变化。通过统计学分析发现,与未处理组相比,500 nmol/L Scriptaid处理15 h囊胚发育率显著增加(P〈0.05),但2细胞阶段分裂率基本不变,而未处理组克隆胚胎在囊胚期Oct4基因表达的倍数明显低于经过Scriptaid处理的克隆胚胎在囊胚期的表达倍数(P〈0.05),这说明经过500 nmol/L Scriptaid处理后,克隆胚胎的Oct4表达量明显提高。未经过处理的克隆胚胎在囊胚期Sox2基因的表达倍数和经过Scriptaid处理的克隆胚胎在囊胚期Sox2基因的表达倍数有差异但是差异不明显,这说明经过500 nmol/L Scriptaid处理之后,基因Sox2表达虽然有提高,但总体来说和未处理的胚胎相比差异不大(P〉0.05)。

Abstract: Histone deacetylases (HDACs) are key proteinases involving in gene regulation. Acetylation is a reversible covalent modification action for protein. HDACs are able to regulate the transcription and expression level of a specific gene and to induce cell differentiation and apoptosis. The aim of the present study was to assess the effect of a novel type of HDACs, Scriptaid, on development of cloned embryos in swine. The cloned pig embryos were treated with different concentrations of Scriptaid for different duration. The cleavage rate and developmental ability of the embryos were determined in vitro. The fetal fibroblasts of landrace pig from Lingyuan Hefeng Pig Breeding Farm were used as donor cells and the embryos were obtained by somatic cell nuclear transfer (SCNT) technology. The reconstructed embryos were treated with different concentrations of Scriptaid (0, 200, 500, 700, 900 nmol/L) for 0, 15, 36, 72 hours, respectively. The cleavage rate and blastocyst development rate of the embryos in two-cell stage were determined. The relative expression of Oct4 and Sox2 genes of the embryos in two-cell stage in untreated group and Scriptaid treatment group were evaluated by using Real-time assay. The results showed that compared to the untreated group, the 500 nmol/L Scriptaid treatment group had significantly higher blastocyst development rate (P〈0.05); no significant differences were observed in cleavage rate of embryos in two-cell stage. In blastocyst stage, the relative expression of Oct4 gene of the cloned embryos in untreated group was significantly lower than that in Scriptaid treatment group (P〈0.05), indicating that the relative expression of Oct4 gene of the cloned embryos was significantly increased after the treatment of 500 nmoFL Scriptaid; no significant differences in the relative expression of Sox2 gene of the cloned embryos between untreated group and Scriptaid treatment group were observed (P〉0.05), indicating that although the relative expression of Oct4 gene was increased after the treatment of 500 nmol/L Scriptaid, there were no significant differences between control group and Scriptaid treatment group.

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