Mitochondrial physiology and gene expression analyses reveal metabolic and translational dysregulation in oocyte-induced somatic nuclear reprogramming.
Mitochondrial physiology and gene expression analyses reveal metabolic and translational dysregulation in oocyte-induced somatic nuclear reprogramming.
复制标题
DOI：
10.1371/journal.pone.0036850
复制
发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Boiani M
中科院分区：
文献类型：
--
作者：
Esteves TC;Psathaki OE;Pfeiffer MJ;Balbach ST;Zeuschner D;Shitara H;Yonekawa H;Siatkowski M;Fuellen G;Boiani M
体细胞核移植（SCNT）后，当对外源核进行重编程时，去核卵母细胞（ooplasm）必须发出信号，表明与核供体细胞相比，生物量和细胞需求发生了变化。利用表达核编码但线粒体靶向的EGFP的细胞，开发了一种直接区分母体和胚胎产物的策略，测试了重编程过程中对转录和转录后活性的卵浆需求。具体来说，我们比较了植入前SCNT胚胎中EGFP和其他产物的转录物和蛋白质水平，并与受精对照（胚胎来自同一卵母细胞池，通过细胞质内注射含有EGFP转基因的精子）。我们观察到，虽然EGFP转录丰度没有差异，但与受精囊胚相比，SCNT中的蛋白质水平显著降低。而Gapdh和Actb则没有，它们的蛋白反映mRNA。这种转录-蛋白关系表明，体细胞核可以跟上卵浆转录的需求，而某些mrna在SCNT后发生转录和翻译错配。我们进一步检测了SCNT后的代谢紊乱，表明在重编程过程中调节转录后变化的力量中有一席之地。我们的观察将卵母细胞诱导的重编程归因于以前未预料到的调节维度，因为功能蛋白的存在可能不再从mRNA推断，而是依赖于可能通过代谢调节的转录后调节。
While reprogramming a foreign nucleus after somatic cell nuclear transfer (SCNT), the enucleated oocyte (ooplasm) must signal that biomass and cellular requirements changed compared to the nucleus donor cell. Using cells expressing nuclear-encoded but mitochondria-targeted EGFP, a strategy was developed to directly distinguish maternal and embryonic products, testing ooplasm demands on transcriptional and post-transcriptional activity during reprogramming. Specifically, we compared transcript and protein levels for EGFP and other products in pre-implantation SCNT embryos, side-by-side to fertilized controls (embryos produced from the same oocyte pool, by intracytoplasmic injection of sperm containing the EGFP transgene). We observed that while EGFP transcript abundance is not different, protein levels are significantly lower in SCNT compared to fertilized blastocysts. This was not observed for Gapdh and Actb, whose protein reflected mRNA. This transcript-protein relationship indicates that the somatic nucleus can keep up with ooplasm transcript demands, whilst transcription and translation mismatch occurs after SCNT for certain mRNAs. We further detected metabolic disturbances after SCNT, suggesting a place among forces regulating post-transcriptional changes during reprogramming. Our observations ascribe oocyte-induced reprogramming with previously unsuspected regulatory dimensions, in that presence of functional proteins may no longer be inferred from mRNA, but rather depend on post-transcriptional regulation possibly modulated through metabolism.
登录
查看更多内容
影响因子：
12.3
作者：
Gentleman RC;Carey VJ;Bates DM;Bolstad B;Dettling M;Dudoit S;Ellis B;Gautier L;Ge Y;Gentry J;Hornik K;Hothorn T;Huber W;Iacus S;Irizarry R;Leisch F;Li C;Maechler M;Rossini AJ;Sawitzki G;Smith C;Smyth G;Tierney L;Yang JY;Zhang J
通讯作者：
Zhang J
影响因子：
2.7
作者：
Balbach, S. T.;Esteves, T. C.;Boiani, M.
通讯作者：
Boiani, M.
影响因子：
2.6
作者：
Cavaleri, Fatima M.;Balbach, Sebastian T.;Boiani, Michele
通讯作者：
Boiani, Michele
影响因子：
29
作者：
Folmes CD;Nelson TJ;Martinez-Fernandez A;Arrell DK;Lindor JZ;Dzeja PP;Ikeda Y;Perez-Terzic C;Terzic A
通讯作者：
Terzic A
影响因子：
8
作者：
Halbeisen, R. E.;Galgano, A.;Scherrer, T.;Gerber, A. P.
通讯作者：
Gerber, A. P.