The developmental potential of mouse somatic cell nuclear-transferred oocytes treated with trichostatin A and 5-aza-2′-deoxycytidine
The developmental potential of mouse somatic cell nuclear-transferred oocytes treated with trichostatin A and 5-aza-2′-deoxycytidine
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DOI：
10.1017/s0967199408005133
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发表时间：
2009-05
期刊：
影响因子：
1.7
通讯作者：
Y. Tsuji;Y. Kato;Y. Tsunoda
中科院分区：
文献类型：
--
作者：
Y. Tsuji;Y. Kato;Y. Tsunoda
总结为了促进核重编程，体细胞或体细胞核移植（SCNT）卵母细胞已用组蛋白脱乙酰酶抑制剂曲古抑菌素A（TSA）或DNA甲基转移酶抑制剂5-氮杂-2 '-脱氧胞苷（5-aza-dC）处理，以放松分化体细胞的表观遗传标记。TSA处理的SCNT卵母细胞具有增加的发育潜力，但最佳处理时期是未知的。体细胞中甲基化水平的降低对SCNT卵母细胞没有积极影响，但尚未研究用5-aza-dC处理SCNT胚胎。我们研究了TSA处理持续时间对小鼠SCNT卵母细胞发育潜力的影响以及5-aza-dC处理对其体外和体内发育潜力的影响。为了确定TSA处理持续时间的影响，将核移植（NT）卵母细胞与100 nM TSA培养0至26 h。用TSA处理8 ~ 12 h的SCNT卵母细胞的囊胚发育率较高，处理8 ~ 12 h后可获得足月胎儿。当卵母细胞处理14 h和26 h时，囊胚率显著降低，未获得胎儿。为了检查5-aza-dC的作用，将2细胞期SCNT胚胎与10或100 nM 5-aza-dC一起培养48小时至桑椹胚期并转移。用5-aza-dC处理的胚胎发育成囊胚的潜力降低，并且在移植后没有获得胎儿。研究结果表明，SCNT小鼠卵母细胞的长期TSA处理和5-aza-dC处理抑制了移植后发育成胚泡和胎儿的潜力。
Summary To facilitate nuclear reprogramming, somatic cells or somatic cell nuclear-transferred (SCNT) oocytes have been treated with the histone deacetylase inhibitor trichostatin A (TSA), or the DNA methyltransferase inhibitor, 5-aza-2′-deoxycytidine (5-aza-dC), to relax epigenetic marks of differentiated somatic cells. TSA-treated SCNT oocytes have increased developmental potential, but the optimal treatment period is unknown. Reduced methylation levels in somatic cells have no positive effect on SCNT oocytes, but the treatment of SCNT embryos with 5-aza-dC has not been investigated. We examined the effect of TSA treatment duration on the developmental potential of mouse SCNT oocytes and the effect of 5-aza-dC treatment on their in vitro and in vivo developmental potential. To determine the effects of TSA treatment duration, nuclear-transferred (NT) oocytes were cultured for 0 to 26 h with 100 nM TSA. SCNT oocytes treated with TSA for 8 to 12 h had the higher rate of development to blastocysts and full-term fetuses were obtained after treatment for 8 to 12 h. When oocytes were treated for 14 h and 26 h, blastocyst rates were significantly decreased and fetuses were not obtained. To examine the effect of 5-aza-dC, 2-cell stage SCNT embryos were cultured with 10 or 100 nM 5-aza-dC for 48 h to the morula stage and transferred. The potential of embryos treated with 5-aza-dC to develop into blastocysts was decreased and no fetuses were obtained after transfer. The findings demonstrated that long-term TSA treatment of SCNT mouse oocytes and treatment with 5-aza-dC inhibit the potential to develop into blastocysts and to fetuses after transfer.