The Effects of Trichostatin A on mRNA Expression of Chromatin Structure-, DNA Methylation-, and Development-Related Genes in Cloned Mouse Blastocysts

Li, Xiangping; Kato, Yuki; Tsuji, Yuta; Tsunoda, Yukio

doi:10.1089/clo.2007.0066

Cited by 67 publications

(51 citation statements)

References 54 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Their dynamic balance regulates gene transcription and gene expression of eukaryotes at the DNA level. Some anomalous expression of HDACs has been observed in SCNT embryos (Suteevun et al 2006;Li et al 2008;Imsoonthornruksa et al 2010;Lee et al 2010). In this study, at the blastocyst stage HDAC2 expression was similar for AMSC-NT embryos and IVF embryos, whereas a lower level was detected in FF-NT embryos.…”

Section: Discussionsupporting

confidence: 51%

Differential developmental competence and gene expression patterns in buffalo (Bubalus bubalis) nuclear transfer embryos reconstructed with fetal fibroblasts and amnion mesenchymal stem cells

Shah

Yadav

2015

Cytotechnology

View full text Add to dashboard Cite

The developmental ability and gene expression pattern at 8-to 16-cell and blastocyst stages of buffalo (Bubalus bubalis) nuclear transfer (NT) embryos from fetal fibroblasts (FFs), amnion mesenchymal stem cells (AMSCs) and in vitro fertilized (IVF) embryos were compared in the present studies. The in vitro expanded buffalo FFs showed a typical ''S'' shape growth curve with a doubling time of 41.4 h and stained positive for vimentin. The in vitro cultured undifferentiated AMSCs showed a doubling time of 39.5 h and stained positive for alkaline phosphatase, and these cells also showed expression of pluripotency markers (OCT4, SOX2, NANOG), and mesenchymal stem cell markers (CD29, CD44) and were negative for haematopoietic marker (CD34) genes at different passages. Further, when AMSCs were exposed to corresponding induction conditions, these cells differentiated into adipogenic, chondrogenic and osteogenic lineages which were confirmed through oil red O, alcian blue and alizarin staining, respectively. Donor cells at 3-4 passage were employed for NT. The cleavage rate was significantly (P \ 0.05) higher in IVF than in FF-NT and AMSC-NT embryos (82.6 ± 8.2 vs. 64.6 ± 1.3 and 72.3 ± 2.2 %, respectively). However, blastocyst rates in IVF and AMSC-NT embryos (30.6 ± 2.7 and 28.9 ± 3.1 %) did not differ and were significantly (P \ 0.05) higher than FF-NT (19.5 ± 1.8 %). Total cell number did not show significant (P [ 0.05) differences between IVF and AMSC-NT embryos (186.7 ± 4.2, 171.2 ± 3.8, respectively) but were significantly (P \ 0.05) higher than that from FF-NT (151.3 ± 4.1). Alterations in the expression pattern of genes implicated in transcription and pluripotency (OCT4, STAT3, NANOG), DNA methylation (DNMT1, DNMT3A), histone deacetylation (HDAC2), growth factor signaling and imprinting (IGF2, IGF2R), apoptosis (BAX, BCL2), metabolism (GLUT1) and oxidative stress (MnSOD) regulation were observed in cloned embryos. The transcripts or expression patterns in AMSC-NT embryos more closely followed that of the in vitro derived embryos compared with FF-NT embryos. The results demonstrate that multipotent amnion MSCs have a greater potential as donor cells than FFs in achieving enhanced production of cloned buffalo embryos.

show abstract

Section: Discussionsupporting

confidence: 51%

Differential developmental competence and gene expression patterns in buffalo (Bubalus bubalis) nuclear transfer embryos reconstructed with fetal fibroblasts and amnion mesenchymal stem cells

Shah

Yadav

2015

Cytotechnology

View full text Add to dashboard Cite

show abstract

“…TSA treatment of SCNT embryos has been reported to be effective for improving the blastocyst formation rate in mice [20][21][22]38] and pigs [23,24]. In contrast, when bovine SCNT embryos were treated with TSA at 50 nM for 13 h [26] and at 5, 50 or 500 nM for 12 h [25] after NT, no significant effects on the development to the blastocyst stage were observed.…”

Section: Discussionmentioning

confidence: 98%

“…One possible explanation is that HDACi treatment of NT embryos may lead to amendment of abnormal epigenetic modifications, followed by normal expression of some development-related genes. Recent reports have demonstrated that TSA treatment of murine NT embryos influences the expression of genes that regulate chromatin structure and DNA methylation at the blastocyst stage [38] and that the histone acetylation levels of TSA-treated NT embryos are more similar to those of in vitro fertilized embryos than those of untreated cloned embryos in cattle [26]. In addition, Shao et al reported that TSA treatment of mouse SCNT embryos enhances the expression of two embryonic genes shown to be transcriptionally active during zygotic gene activation (ZGA) [39].…”

Section: Discussionmentioning

confidence: 99%

Treatment with a Histone Deacetylase Inhibitor after Nuclear Transfer Improves the Preimplantation Development of Cloned Bovine Embryos

Akagi

Matsukawa

Mizutani

et al. 2011

Journal of Reproduction and Development

View full text Add to dashboard Cite

Abstract. We examined the effects of treatment with histone deacetylase inhibitors (HDACi), trichostatin A (TSA) and scriptaid (SCR), on the blastocyst formation rate in bovine somatic cell nuclear transferred (SCNT) embryos derived from fibroblast cells. Three fibroblast cell lines (L1, L2 and L3) were used as somatic cell donors to produce SCNT embryos (L1, L2 and L3 embryos, respectively). In Experiment 1, we compared the in vitro developmental competence of L1 embryos treated with various concentrations of TSA for different time periods following chemical activation. Embryos treated with 5 nM TSA for 20 h showed a significantly increased blastocyst formation rate compared with untreated controls. In Experiment 2, we examined the effect of TSA (5 nM) treatment of L1, L2 and L3 embryos as well as the effect of treatment of L1, L2 and L3 embryos with various concentrations of SCR on in vitro developmental competence. It was found that 5 nM TSA treatment significantly increased the blastocyst formation rate in L1 and L3 embryos but did not have an influence on the development of L2 embryos. On the other hand, 5 nM SCR treatment significantly increased the blastocyst formation rates of L1 and L2 embryos compared with controls. However, there was no significant increase in the blastocyst formation rate of L3 embryos when they were treated with SCR. In Experiment 3, acetylation of H4K12 was examined in donor cells and pronuclear-stage L1, L2 and L3 embryos treated with 5 nM TSA or 5 nM SCR by immunostaining. The level of H4K12 acetylation was different among donor cells. The staining intensities in the TSA-treated L1 and L3 embryos and SCR-treated L2 embryos were significantly higher than those of untreated embryos. These results suggest that HDACi treatment of bovine SCNT embryos improves the blastocyst formation rate; however, the optimal treatment conditions may differ among donor cell lines. Key words: Bovine, Nuclear transfer, Histone deacetylase inhibitor (J. Reprod. Dev. 57: [120][121][122][123][124][125][126] 2011) ive cloned offspring have been produced in many species by somatic cell nuclear transfer (SCNT), although the cloning efficiency is still very low. It has been reported that, in SCNT, embryo losses occur at various stages, and the anomalies in fetal and perinatal stages have been observed [1][2][3][4][5][6]. Because such abnormalities of clones are not transmitted to their progenies [7][8][9][10][11][12], most of the developmental problems of clones are believed to be the results of epigenetic defects [13]. In fact, several studies have revealed abnormal epigenetic modifications such as DNA methylation and histone modifications in SCNT embryos [14][15][16][17].A wide variety of histone deacetylase inhibitors (HDACi) of both natural and synthetic origin has been reported [18]. Trichostatin A (TSA), which is a natural product isolated from Streptomyces hygroscopicus [19], is one of the most frequently used drugs for various studies as a selective HDACi. Recently, it has been reported from two la...

show abstract

“…Therefore, one of the reasons for the improved developmental competence of SCNT embryos with TSA treatment may be the access of reprogramming-related factors to nucleosomes. In addition, it has been shown that TSA treatment induces not only histone acetylation but also DNA demethylation [39,40]. In fact, Ding et al [29] suggested that TSA treatment induces a higher level of histone acetylation and lower level of DNA methylation at the 2-cell stage in bovine SCNT embryos, which facilitates epigenetic reprogramming of the transferred somatic cell nucleus.…”

Section: Histone Acetylation Of Miniature Pig Scnt Embryosmentioning

confidence: 99%

Acetylation Level of Histone H3 in Early Embryonic Stages Affects Subsequent Development of Miniature Pig Somatic Cell Nuclear Transfer Embryos

Yamanaka

Sugimura

Wakai³

et al. 2009

J. Reprod. Dev.

View full text Add to dashboard Cite

Abstract.Successful cloning by somatic cell nuclear transfer (SCNT) requires a reprogramming process in which the epigenetic state of a differentiated donor nucleus must be converted into an embryonic totipotent state. However, this epigenetic reprogramming is incomplete in SCNT embryos, causing low production efficiency. Recently, it has been reported that trichostatin A (TSA), an inhibitor of histone deacetylase, potentially enhances cloning efficiency. The aim of the present study was to optimize the TSA treatment for miniature pig SCNT embryos and investigate the effect of the acetylation level of histone on developmental competence of SCNT embryos. In order to optimize the TSA treatment, we examined the developmental competence of SCNT embryos under various exposure times (0-50 h) and concentrations (0-500 nM). Treatment with 5 nM TSA for 15 and 20 h beginning at the start of activation significantly increased the blastocyst formation rate (34.6 and 32.4 vs. 18.2%, respectively) and mean cell number (57.0 ± 2.7 and 56.6 ± 2.7 vs. 43.5 ± 2.1, respectively) as compared with the non-treated group (0 h). We then investigated the acetylation levels of histone H3 in SCNT embryos treated with or without TSA (TSA (+) or TSA (-)) as compared with in vitrofertilized (IVF) embryos. The acetylation levels of the TSA (-) SCNT embryos at the pseudo-pronuclear and 2-cell stages were significantly lower than those of the IVF embryos at the same developmental stages. In contrast, the acetylation levels of the TSA (+) SCNT embryos were similar to those of the IVF embryos. There was no difference in the acetylation levels of all groups at the blastocyst stage. Our data therefore suggests that the acetylation level of histone H3 at the pseudo-pronuclear and 2-cell stages is positively correlated with subsequent development of SCNT embryos, which may be an important event for the vital development of SCNT embryos in miniature pigs. Key words: Embryo development, Histone acetylation, Miniature pig, Nuclear transfer, Trichostatin A (J. Reprod. Dev. 55: [638][639][640][641][642][643][644] 2009) lthough many studies have been performed to improve the developmental competence of miniature pig SCNT embryos [1][2][3][4], the overall efficiency remains low. To obtain sufficient developmental competence for SCNT embryos to develop to term, the differentiated cell nucleus should be subjected to epigenetic reprogramming processes including chromatin remodeling and DNA methylation during preimplantation development. However, reprogramming of a differentiated nucleus to an embryonic state is reportedly delayed and incomplete in SCNT embryos [5]. In fact, previous studies have demonstrated that epigenetic modifications such as DNA methylation and histone acetylation are abnormally reprogrammed in SCNT embryos during early development [6,7]. Incomplete reprogramming of epigenetic modifications causes abnormal gene expression including imprinted genes [8], resulting in deleterious effects on the development of SCNT embryos [9].Histone acety...

show abstract

The Effects of Trichostatin A on mRNA Expression of Chromatin Structure-, DNA Methylation-, and Development-Related Genes in Cloned Mouse Blastocysts

Cited by 67 publications

References 54 publications

Differential developmental competence and gene expression patterns in buffalo (Bubalus bubalis) nuclear transfer embryos reconstructed with fetal fibroblasts and amnion mesenchymal stem cells

Differential developmental competence and gene expression patterns in buffalo (Bubalus bubalis) nuclear transfer embryos reconstructed with fetal fibroblasts and amnion mesenchymal stem cells

Treatment with a Histone Deacetylase Inhibitor after Nuclear Transfer Improves the Preimplantation Development of Cloned Bovine Embryos

Acetylation Level of Histone H3 in Early Embryonic Stages Affects Subsequent Development of Miniature Pig Somatic Cell Nuclear Transfer Embryos

Contact Info

Product

Resources

About