Activation of bovine somatic cell nuclear transfer embryos by PLCZ cRNA injection.
Activation of bovine somatic cell nuclear transfer embryos by PLCZ cRNA injection.
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DOI：
10.1530/rep-08-0419
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发表时间：
2009-03
期刊：
影响因子：
3.8
通讯作者：
P. Ross;R. Rodríguez;A. E. Iager;Z. Beyhan;Kai Wang;N. Ragina;Sook‐Young Yoon;R. Fissore;J. Cibelli
中科院分区：
文献类型：
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作者：
P. Ross;R. Rodríguez;A. E. Iager;Z. Beyhan;Kai Wang;N. Ragina;Sook‐Young Yoon;R. Fissore;J. Cibelli
通过将分化的体细胞转移到去核的卵母细胞中来生产克隆动物，从而避免了受精。在受精过程中，精子释放一种精子特异性磷脂酶C（PLCZ），负责触发Ca（2）（+）振荡和卵母细胞激活。在牛体细胞核移植（SCNT）过程中，卵母细胞活化是通过联合化学处理人工实现的，这些化学处理诱导细胞内Ca（2）（+）单调上升并抑制磷酸化或蛋白质合成。在这项研究中，我们测试的假设，激活牛核移植胚胎PLCZ提高核重编程。将PLCZ cRNA注射到牛SCNT单位中诱导了类似于受精后观察到的Ca（2）（+）振荡，并支持了类似于IVF产生的胚胎中观察到的高囊胚发育率。此外，在八细胞和囊胚阶段的基因表达分析揭示了两组胚胎中一些基因的相似表达模式。最后，水平的三甲基化赖氨酸27在组蛋白H3的囊胚中较高的牛核移植胚胎使用放线菌酮和6-二甲基氨基嘌呤（DMAP）激活比那些使用PLCZ激活或来自IVF。这些结果表明，外源性PLCZ可用于激活牛SCNT衍生的胚胎，并支持受精样激活反应可以增强核重编程的某些方面的假设。
The production of cloned animals by the transfer of a differentiated somatic cell into an enucleated oocyte circumvents fertilization. During fertilization, the sperm delivers a sperm-specific phospholipase C (PLCZ) that is responsible for triggering Ca(2)(+) oscillations and oocyte activation. During bovine somatic cell nuclear transfer (SCNT), oocyte activation is artificially achieved by combined chemical treatments that induce a monotonic rise in intracellular Ca(2)(+) and inhibit either phosphorylation or protein synthesis. In this study, we tested the hypothesis that activation of bovine nuclear transfer embryos by PLCZ improves nuclear reprogramming. Injection of PLCZ cRNA into bovine SCNT units induced Ca(2)(+) oscillations similar to those observed after fertilization and supported high rates of blastocyst development similar to that seen in embryos produced by IVF. Furthermore, gene expression analysis at the eight-cell and blastocyst stages revealed a similar expression pattern for a number of genes in both groups of embryos. Lastly, levels of trimethylated lysine 27 at histone H3 in blastocysts were higher in bovine nuclear transfer embryos activated using cycloheximide and 6-dimethylaminopurine (DMAP) than in those activated using PLCZ or derived from IVF. These results demonstrate that exogenous PLCZ can be used to activate bovine SCNT-derived embryos and support the hypothesis that a fertilization-like activation response can enhance some aspects of nuclear reprogramming.