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曲古抑菌素A改善牛克隆胚胎的植入前发育并改变克隆囊胚中表观遗传和多能性基因的表达。

Trichostatin a improves preimplantation development of bovine cloned embryos and alters expression of epigenetic and pluripotency genes in cloned blastocysts.

作者信息

Oh Hyun Ju, Lee Tae Hee, Lee Ji Hyun, Lee Byeong Chun

机构信息

Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul National University, Seoul, Korea.

出版信息

J Vet Med Sci. 2012 Nov;74(11):1409-15. doi: 10.1292/jvms.11-0510. Epub 2012 Jun 18.

Abstract

We investigated the effects of exposure time and concentration of trichostatin A (TSA) on in vitro development and quality of bovine SCNT embryos. At multiple time points, the relative expression of genes related to pluripotency and reprogramming was analyzed in order to assess the quality of preimplantation embryos cultured in media with TSA using real-time PCR. Development into blastocysts was higher in 100 nM TSA than in controls (35.96 vs. 28.30%, P<0.05). Study of 100 nM TSA exposure time showed development into blastocysts was higher during both short-term and long-term exposure than in controls (36.17 and 34.04 vs. 23.45%), but there was no significant difference between TSA groups. Nanog expression in blastocysts after long-term TSA exposure was similar to that in IVF blastocysts and greater than in controls and short-term exposed embryos. The Oct4 levels in the short-term exposure group were similar to those of IVF blastocysts, while Oct4 expression in long-term exposed embryos was significantly higher than in other groups. Measurement of DNMT1 and HDAC1 in blastocysts showed a similar expression profile among IVF and TSA groups regardless of treatment duration. In conclusion, this study suggests that TSA treatment after SCNT in bovine embryos can improve in vitro development of embryos by increasing the blastocysts formation and positive reprogramming of the reconstructed embryo genome caused by downregulation of DNA methylation and up-regulation of pluripotency.

摘要

我们研究了曲古抑菌素A(TSA)的暴露时间和浓度对牛体细胞核移植(SCNT)胚胎体外发育及质量的影响。在多个时间点,使用实时定量PCR分析了与多能性和重编程相关基因的相对表达,以评估在含TSA的培养基中培养的植入前胚胎的质量。100 nM TSA组胚胎发育为囊胚的比例高于对照组(35.96%对28.30%,P<0.05)。对100 nM TSA暴露时间的研究表明,短期和长期暴露期间发育为囊胚的比例均高于对照组(36.17%和34.04%对23.45%),但TSA组之间无显著差异。长期TSA暴露后囊胚中的Nanog表达与体外受精(IVF)囊胚相似,且高于对照组和短期暴露胚胎。短期暴露组的Oct4水平与IVF囊胚相似,而长期暴露胚胎中的Oct4表达显著高于其他组。对囊胚中DNMT1和HDAC1的检测显示,无论处理时间长短,IVF组和TSA组之间的表达谱相似。总之,本研究表明,牛胚胎SCNT后进行TSA处理可通过增加囊胚形成以及由DNA甲基化下调和多能性上调引起的重构胚胎基因组的正向重编程来改善胚胎的体外发育。

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