Although the somatic cell nuclear transfer(SCNT) technique has been used extensively for cloning and generating transgenic pigs,the cloning efficiency is still very low.It has been proposed that the low efficiency of ...Although the somatic cell nuclear transfer(SCNT) technique has been used extensively for cloning and generating transgenic pigs,the cloning efficiency is still very low.It has been proposed that the low efficiency of this technique is the result of incomplete epigenetic reprogramming and abnormal gene expression during early embryonic development.In this study,we investigate the effect of Scriptaid,a low-toxicity histone deacetylase inhibitor,on the developmental competence of porcine SCNT embryos.We found that treating SCNT embryos with 500 nmol/L Scriptaid for 15 h after activation significantly enhanced the blastocyst formation rate(27.7%) compared with the untreated group(control)(12.2%,P<0.05).Using an immunofluorescence technique to measure the average fluorescence intensity,we also found that treating SCNT embryos with Scriptaid increased the level of histone acetylation on histone H3 at lysine 14(acH3K14).Furthermore,treating embryos with Scriptaid increased the expression level of three genes that play important roles during embryonic development(Oct4,Klf4 at the blastocyst stage and Nanog at the 4-cell stage).Moreover,the expression level of the apoptosis-related gene Caspase-3 was significantly lower in the Scriptaid-treated SCNT embryos compared with the control SCNT embryos at the 4-cell and blastocyst stages.In conclusion,these results indicate that Scriptaid treatment improves the development and nuclear reprogramming of porcine SCNT embryos.展开更多
基金This work was supported by the National Basic Research Program of China （2009CB941000）.
文摘Although the somatic cell nuclear transfer(SCNT) technique has been used extensively for cloning and generating transgenic pigs,the cloning efficiency is still very low.It has been proposed that the low efficiency of this technique is the result of incomplete epigenetic reprogramming and abnormal gene expression during early embryonic development.In this study,we investigate the effect of Scriptaid,a low-toxicity histone deacetylase inhibitor,on the developmental competence of porcine SCNT embryos.We found that treating SCNT embryos with 500 nmol/L Scriptaid for 15 h after activation significantly enhanced the blastocyst formation rate(27.7%) compared with the untreated group(control)(12.2%,P<0.05).Using an immunofluorescence technique to measure the average fluorescence intensity,we also found that treating SCNT embryos with Scriptaid increased the level of histone acetylation on histone H3 at lysine 14(acH3K14).Furthermore,treating embryos with Scriptaid increased the expression level of three genes that play important roles during embryonic development(Oct4,Klf4 at the blastocyst stage and Nanog at the 4-cell stage).Moreover,the expression level of the apoptosis-related gene Caspase-3 was significantly lower in the Scriptaid-treated SCNT embryos compared with the control SCNT embryos at the 4-cell and blastocyst stages.In conclusion,these results indicate that Scriptaid treatment improves the development and nuclear reprogramming of porcine SCNT embryos.
基金This work was supported by the grants from the Special Fund of Doctor Station of University of Education Department of China （no. 200802260007）, the Innovation Project of Main Instructor of Education Department of Heilongjiang Province, China （no. 1054G024）, and the Important Project of Overseas Scholar of Education Department of Heilongjiang Province, China （no. 1152hq34）.
文摘这研究的目的是在老鼠由一本高红花油食谱和 adipocytic 基因表示调查导致食谱的肥胖的预防。四十只 3-week-old C57BL/6 老鼠随机被划分成三个组：控制组(反对， 5% 猪油 +5% 红花油) ，高猪油组(LAR， 45% 猪油 +5% 红花油) ，并且高红花油组(SAF， 45% 红花油 +5% 猪油) 。在 10 个星期以后， LAR 组的 10 只老鼠高被换到红花油饮食(LARSAF ) 。十个星期以后，葡萄糖忍耐测试被葡萄糖的 intraperitoneal 注射执行。类脂化合物和胰岛素的传播层次被测量，白脂肪质的纸巾为基因薄片和反向的 transcriptasepolymerase 链反应分析被拿。LAR 组比 CON 组显示出更高的身体重量，脂肪过多索引，胰岛素，和类脂化合物(P < 0.05 ) 。在 LARSAF 组的身体重量在饮食的颠倒以后减少了。血浆生物化学的侧面在 LARSAF 和 SAF 组减少了(P < 0.05 ) 与那些相比 LAR 组。LARSAF 组的血葡萄糖水平与 LAR 组的相比在 intraperitoneal 葡萄糖忍耐测试期间被减少。LARSAF 组有 Orexin 和 Ghrelin 基因表示的底层，而 PPAR 基因表示的水平显著地与 LAR 组的相比被提高。那么， SAF 食谱能改变 adipocytic 在导致食谱的肥胖的有效改善的脂肪过多相关的基因表示和结果。
