Alizadeh Zohreh, Kageyama Shun-Ichiro, Aoki Fugaku
Department of Integrated Biosciences, Graduate School of Frontier Sciences, University of Tokyo, Chiba, Japan.
Mol Reprod Dev. 2005 Nov;72(3):281-90. doi: 10.1002/mrd.20340.
During oogenesis, mRNA is actively transcribed and accumulated in growing oocytes, but this transcription stops before the oocytes grow to their full size. The accumulated maternal mRNA is used for protein synthesis in the oocytes during meiotic maturation and even in the embryos to sustain development after fertilization. Therefore, the degradation of accumulated maternal mRNA starts during meiotic maturation, but its rate is slow. Nevertheless, some mRNA species should rapidly degrade after fertilization if they encode proteins that play a role in specific events during meiosis and are detrimental for development after fertilization. In this study, to identify the selective degradation of maternal transcripts after fertilization, we sought mRNAs that are degraded in the early hours after fertilization by constructing an oocyte cDNA library after subtracting the cDNA of embryos at the mid one-cell stage. H1oo, c-mos, tPA (tissue type plasminogen activator gene), and Gdf9 were identified as genes whose transcripts undergo rapid degradation after fertilization. RT-PCR analysis showed that none of these transcripts was expressed during pre-implantation development once they were eliminated, suggesting that the mRNA species that are required for oogenesis, but not for early pre-implantation development, are degraded rapidly after fertilization. Microinjection of chimeric mRNAs in which the coding and 3'-untranslated regions (3'UTR) were exchanged between c-mos and hypoxanthine phosphoribosyltransferase mRNAs revealed that the 3'UTR plays a role in the rapid degradation that occurs after fertilization. Cytoplasmic polyadenylation elements (CPEs) was found near a poly(A) signal in the 3'UTR of all the mRNA species identified as rapidly degrading mRNA. The mechanism for the selective degradation is discussed, in relation to its biological significance.
在卵子发生过程中,mRNA在生长中的卵母细胞中被积极转录和积累,但这种转录在卵母细胞生长到其完整大小之前就停止了。积累的母体mRNA在减数分裂成熟期间用于卵母细胞中的蛋白质合成,甚至在胚胎中用于维持受精后的发育。因此,积累的母体mRNA的降解在减数分裂成熟期间开始,但其速率较慢。然而,如果某些mRNA物种编码在减数分裂期间特定事件中起作用且对受精后发育有害的蛋白质,则它们在受精后应迅速降解。在本研究中,为了鉴定受精后母体转录本的选择性降解,我们通过构建减去单细胞中期胚胎cDNA后的卵母细胞cDNA文库,寻找在受精后早期被降解的mRNA。H1oo、c-mos、tPA(组织型纤溶酶原激活剂基因)和Gdf9被鉴定为其转录本在受精后经历快速降解的基因。RT-PCR分析表明,一旦这些转录本被消除,它们在植入前发育期间均未表达,这表明卵子发生所需但早期植入前发育不需要的mRNA物种在受精后迅速降解。将编码区和3'-非翻译区(3'UTR)在c-mos和次黄嘌呤磷酸核糖基转移酶mRNA之间交换的嵌合mRNA进行显微注射,结果表明3'UTR在受精后发生的快速降解中起作用。在所有被鉴定为快速降解mRNA的mRNA物种的3'UTR中的聚腺苷酸信号附近发现了细胞质聚腺苷酸化元件(CPE)。本文讨论了选择性降解的机制及其生物学意义。