Insufficient epigenetic reprogramming of donor nuclei is believed to be one of the most important causes of low development efficiency of mammalian somatic cell nuclear transfer （SCNT）. Previous studies have shown t...Insufficient epigenetic reprogramming of donor nuclei is believed to be one of the most important causes of low development efficiency of mammalian somatic cell nuclear transfer （SCNT）. Previous studies have shown that both the in vitro and in vivo development of mouse SCNT embryos could be increased significantly by treat- ment with various histone deacetylase inhibitors （HDACi）, including Trichostatin A, Scriptaid, and m-carboxycin- namic acid bishydroxamide （CBHA）, in which only the effect of CBHA has not yet been tested in other species. In this paper we examine the effect of CBHA treatment on the development of porcine SCNT embryos. We have dis- covered the optimum dosage and time for CBHA treat- ment： incubating SCNT embryos with 2 pmol/L CBHA for 24 h after activation could increase the blastocyst rate from 12.7% to 26.5%. Immunofluorescence results showed that the level of acetylation at histone 3 lysine 9 （AcH3K9）, acetylation at histone 3 lysine 18 （AcH3K18）, and acetylation at histone 4 lysine 16 （AcH4K16） was raised after CBHA treatment. Meanwhile, CBHA treatment improved the expression of development relating genes such as pou5fl, cdx2, and the imprinted genes like igf2. Despite these promising in vitro results and histone reprogramming, the full term development was notsignificantly increased after treatment. In conclusion, CBHA improves the in vitro development of pig SCNT embryos, increases the global histone acetylation and corrects the expression of some developmentally important genes at early stages. As in mouse SCNT, we have shown that nuclear epigenetic reprogramming in pig early SCNT embryos can be modified by CBHA treatment.展开更多
Over 50 years of efforts, cellular reprogram- ruing opens a new door for disease modeling and regen- erative medicine. Although induction of pluripotency by transcription factors has become common, only a small portio...Over 50 years of efforts, cellular reprogram- ruing opens a new door for disease modeling and regen- erative medicine. Although induction of pluripotency by transcription factors has become common, only a small portion of basic mechanisms of epigenetic modifications during this process have been revealed. To clearly under- stand reprogramming and devise ways to promote full transition towards pluripotency, we must gain insight from comprehensive characterizations of cells at distinct repro- gramming stages, which involves gene expression profil- ing, chromatin state maps of key activating and repressive marks, and DNA modifications. Here, we review recent advances in epigenetic reprogramming to pluripotency with a focus on the principal molecular regulators and attach importance to the combination of high-throughput sequencing and systematic biology approaches in uncov- ering underlying molecular mechanisms of this unique platform in future researches.展开更多
文摘Insufficient epigenetic reprogramming of donor nuclei is believed to be one of the most important causes of low development efficiency of mammalian somatic cell nuclear transfer （SCNT）. Previous studies have shown that both the in vitro and in vivo development of mouse SCNT embryos could be increased significantly by treat- ment with various histone deacetylase inhibitors （HDACi）, including Trichostatin A, Scriptaid, and m-carboxycin- namic acid bishydroxamide （CBHA）, in which only the effect of CBHA has not yet been tested in other species. In this paper we examine the effect of CBHA treatment on the development of porcine SCNT embryos. We have dis- covered the optimum dosage and time for CBHA treat- ment： incubating SCNT embryos with 2 pmol/L CBHA for 24 h after activation could increase the blastocyst rate from 12.7% to 26.5%. Immunofluorescence results showed that the level of acetylation at histone 3 lysine 9 （AcH3K9）, acetylation at histone 3 lysine 18 （AcH3K18）, and acetylation at histone 4 lysine 16 （AcH4K16） was raised after CBHA treatment. Meanwhile, CBHA treatment improved the expression of development relating genes such as pou5fl, cdx2, and the imprinted genes like igf2. Despite these promising in vitro results and histone reprogramming, the full term development was notsignificantly increased after treatment. In conclusion, CBHA improves the in vitro development of pig SCNT embryos, increases the global histone acetylation and corrects the expression of some developmentally important genes at early stages. As in mouse SCNT, we have shown that nuclear epigenetic reprogramming in pig early SCNT embryos can be modified by CBHA treatment.
基金supported by the National Natural Science Foundation of China(31325019,91319306 and31401247)Ministry of Science and Technology of China(2015CB964800 and 2014CB964601)
文摘Over 50 years of efforts, cellular reprogram- ruing opens a new door for disease modeling and regen- erative medicine. Although induction of pluripotency by transcription factors has become common, only a small portion of basic mechanisms of epigenetic modifications during this process have been revealed. To clearly under- stand reprogramming and devise ways to promote full transition towards pluripotency, we must gain insight from comprehensive characterizations of cells at distinct repro- gramming stages, which involves gene expression profil- ing, chromatin state maps of key activating and repressive marks, and DNA modifications. Here, we review recent advances in epigenetic reprogramming to pluripotency with a focus on the principal molecular regulators and attach importance to the combination of high-throughput sequencing and systematic biology approaches in uncov- ering underlying molecular mechanisms of this unique platform in future researches.
