Zimmermann J W, Schultz R M
Department of Biology, University of Pennsylvania, Philadelphia 19104-6018.
Proc Natl Acad Sci U S A. 1994 Jun 7;91(12):5456-60. doi: 10.1073/pnas.91.12.5456.
The analysis of differential gene expression during preimplantation embryogenesis has been hindered by the paucity of biological material. We report modifications of the recently described mRNA differential display method (Liang, P. & Pardee, A. B. (1992) Science 257, 967-971) to analyze differential gene expression during mouse preimplantation development. The method detects the appropriate changes in the temporal pattern of expression of an amplicon that by DNA sequence analysis is the cytokeratin endo A, a gene whose temporal pattern of expression has been previously determined by S1 nuclease digestion. In addition, this method identifies amplicons that likely represent genes (i) that encode maternal mRNAs, (ii) that are products of early and late zygotic gene activation, (iii) whose expression is greatest during the eight-cell stage (i.e., expressed in a stage-specific manner), and (iv) whose expression is greatest in the blastocyst. In addition to endo A, sequence analysis of these amplicons reveals that an amplicon that displays a temporal pattern of expression consistent with it being a maternal mRNA is the alpha subunit of the mitochondrial F1 ATP synthase.
植入前胚胎发生过程中差异基因表达的分析一直受到生物材料匮乏的阻碍。我们报告了对最近描述的mRNA差异显示方法(Liang, P. & Pardee, A. B. (1992) Science 257, 967 - 971)的改进,以分析小鼠植入前发育过程中的差异基因表达。该方法检测到一个扩增子表达时间模式的适当变化,通过DNA序列分析确定该扩增子为细胞角蛋白内肽酶A,其表达时间模式先前已通过S1核酸酶消化确定。此外,该方法还鉴定出可能代表以下基因的扩增子:(i)编码母体mRNA的基因;(ii)早期和晚期合子基因激活产物的基因;(iii)在八细胞阶段表达量最高(即以阶段特异性方式表达)的基因;(iv)在囊胚中表达量最高的基因。除了内肽酶A,对这些扩增子的序列分析表明,一个显示出与母体mRNA一致的表达时间模式的扩增子是线粒体F1 ATP合酶的α亚基。