Su You-Qiang, Sugiura Koji, Woo Yong, Wigglesworth Karen, Kamdar Sonya, Affourtit Jason, Eppig John J
The Jackson Laboratory, Bar Harbor, ME 04609, USA.
Dev Biol. 2007 Feb 1;302(1):104-17. doi: 10.1016/j.ydbio.2006.09.008. Epub 2006 Sep 12.
There is massive destruction of transcripts during the maturation of mouse oocytes. The objective of this project was to identify and characterize the transcripts that are degraded versus those that are stable during the transcriptionally silent germinal vesicle (GV)-stage to metaphase II (MII)-stage transition using a microarray approach. A system for oocyte transcript amplification using both internal and 3'-poly(A) priming was utilized to minimize the impact of complex variations in transcript polyadenylation prevalent during this transition. Transcripts were identified and quantified using the Affymetrix Mouse Genome 430 v2.0 GeneChip. The significantly changed and stable transcripts were analyzed using Ingenuity Pathways Analysis and GenMAPP/MAPPFinder to characterize the biological themes underlying global changes in oocyte transcripts during maturation. It was concluded that the destruction of transcripts during the GV to MII transition is a selective rather than promiscuous process in mouse oocytes. In general, transcripts involved in processes that are associated with meiotic arrest at the GV-stage and the progression of oocyte maturation, such as oxidative phosphorylation, energy production, and protein synthesis and metabolism, were dramatically degraded. In contrast, transcripts encoding participants in signaling pathways essential for maintaining the unique characteristics of the MII-arrested oocyte, such as those involved in protein kinase pathways, were the most prominent among the stable transcripts.
在小鼠卵母细胞成熟过程中存在大量转录本的降解。本项目的目的是使用微阵列方法,鉴定并表征在转录沉默的生发泡(GV)期至中期II(MII)期转变过程中被降解的转录本与稳定的转录本。利用一种同时使用内部引物和3'-聚腺苷酸化引物的卵母细胞转录本扩增系统,以尽量减少在此转变过程中普遍存在的转录本聚腺苷酸化复杂变化的影响。使用Affymetrix小鼠基因组430 v2.0基因芯片鉴定并定量转录本。使用Ingenuity Pathways Analysis和GenMAPP/MAPPFinder分析显著变化和稳定的转录本,以表征卵母细胞转录本在成熟过程中整体变化背后的生物学主题。得出的结论是,在小鼠卵母细胞中,从GV期到MII期的转变过程中转录本的降解是一个选择性而非杂乱无章的过程。一般来说,参与与GV期减数分裂停滞和卵母细胞成熟进程相关过程的转录本,如氧化磷酸化、能量产生以及蛋白质合成和代谢,会被显著降解。相比之下,编码参与维持MII期停滞卵母细胞独特特征所必需的信号通路的转录本,如参与蛋白激酶通路的转录本,是稳定转录本中最突出的。