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利用cDNA微阵列分析对四倍体小鼠胚胎发育阶段进行表达谱分析。

Expression profiling of tetraploid mouse embryos in the developmental stages using a cDNA microarray analysis.

作者信息

Kawaguchi Jutaro, Kano Kiyoshi, Naito Kunihiko

机构信息

Laboratory of Applied Genetics, Graduate School of Agricultural and Life Science, University of Tokyo, Tokyo, Japan.

出版信息

J Reprod Dev. 2009 Dec;55(6):670-5. doi: 10.1262/jrd.09-127a. Epub 2009 Sep 29.

Abstract

Tetraploid embryos normally develop to the blastocyst stage before implantation, but fail to survive after implantation. To better understand these characteristics of the tetraploid embryo, we produced tetraploid embryos by electrofusion and analyzed expressed genes that participated in mammalian embryogenesis using a DNA microarray analysis and a publicly available bioinformatics analysis of hatched tetraploid and diploid blastocysts. Transcriptome analysis with the DNA microarray revealed that the expression level of most genes was almost the same between diploid and tetraploid blastocysts. We found that the expression levels 2,800 genes were increased, but the expression levels over 1,600 genes were decreased in tetraploid blastocysts, which have a genomic composition identical to that of diploid blastocysts. In tetraploid blastocysts, the levels of 15 genes were decreased more than two-fold compared with the levels in diploid blastocysts. Among these downregulated genes, Ccnb1 (cyclin B1), which was decreased by 3-fold, seemed to play a particularly important role in the cellular organization of the tetraploid blastocyst. To classify the major functional classes of all the genes differentially expressed between diploid and tetraploid blastocysts, we employed a publicly available bioinformatics database, the VisuaL Annotation Display (VLAD). VLAD revealed several altered pathways in tetraploid blastocysts. Some of the enhanced biological processes were moderately involved with chromosome organization, while the suppressed processes were significantly involved with cell division and the mitotic cell cycle, metabolic processes and protein localization and transport. Taken together, our results revealed a large population of downregulated genes in tetraploid hatched blastocysts, and our convergent data suggest that the downregulation might primarily individualize the tetraploid phenotype in hatched blastocysts.

摘要

四倍体胚胎通常在着床前发育到囊胚阶段,但在着床后无法存活。为了更好地了解四倍体胚胎的这些特征,我们通过电融合产生了四倍体胚胎,并使用DNA微阵列分析以及对孵化后的四倍体和二倍体囊胚进行公开可用的生物信息学分析,来分析参与哺乳动物胚胎发生的表达基因。DNA微阵列的转录组分析显示,二倍体和四倍体囊胚之间大多数基因的表达水平几乎相同。我们发现,在具有与二倍体囊胚相同基因组组成的四倍体囊胚中,2800个基因的表达水平升高,但超过1600个基因的表达水平降低。在四倍体囊胚中,15个基因的水平与二倍体囊胚相比降低了两倍以上。在这些下调的基因中,Ccnb1(细胞周期蛋白B1)降低了3倍,似乎在四倍体囊胚的细胞组织中发挥着特别重要的作用。为了对二倍体和四倍体囊胚之间差异表达的所有基因的主要功能类别进行分类,我们使用了一个公开可用的生物信息学数据库,即视觉注释显示(VLAD)。VLAD揭示了四倍体囊胚中的几种改变的途径。一些增强的生物学过程适度参与染色体组织,而受抑制的过程则显著参与细胞分裂和有丝分裂细胞周期、代谢过程以及蛋白质定位和运输。综上所述,我们的结果揭示了四倍体孵化囊胚中大量下调的基因,并且我们的汇总数据表明,这种下调可能主要使孵化囊胚中的四倍体表型个体化。

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