Latham K E, Garrels J I, Chang C, Solter D
Wistar Institute of Anatomy and Biology, Philadelphia, Pennsylvania 19104.
Appl Theor Electrophor. 1992;2(6):163-70.
Numerous studies have revealed stage specific alterations in protein synthesis that occur in mouse embryos. A thorough analysis of these changes has been hampered by limitations in the ability to resolve individual proteins, the ability to accurately quantify and manage data from two-dimensional gel images, and by variation in the staging of the embryos used. To learn more of the changes in protein synthesis that occur during early development, we constructed a protein database for the mouse embryo using the QUEST system of high-resolution, two-dimensional gel electrophoresis and computerized gel image analysis (Garrels, 1989, J. Biol. Chem., 264:526). Synchronous cohorts of embryos were labeled at 3 h intervals throughout the entire preimplantation period from fertilization to blastocyst stage in order to characterize in detail the changes in protein synthesis pattern that occur during normal preimplantation development. Additional samples were prepared from early post-implantation embryos, isolated inner cell mass and trophoblast cells, and cultured embryonic stem cells. These provide the means for identifying cell and tissue specific proteins and for characterizing changes in protein synthesis that accompany early cellular differentiation in the embryo. We present here a description of the mouse embryo database and discuss its potential usefulness to the study of mammalian embryogenesis.
众多研究揭示了小鼠胚胎中发生的蛋白质合成的阶段特异性变化。对这些变化进行全面分析受到多种限制,包括分辨单个蛋白质的能力、准确量化和管理二维凝胶图像数据的能力,以及所使用胚胎分期的差异。为了更多地了解早期发育过程中发生的蛋白质合成变化,我们使用高分辨率二维凝胶电泳和计算机凝胶图像分析的QUEST系统(Garrels,1989,《生物化学杂志》,264:526)构建了小鼠胚胎蛋白质数据库。在从受精到囊胚期的整个植入前阶段,以3小时的间隔对同步胚胎群体进行标记,以便详细表征正常植入前发育过程中发生的蛋白质合成模式变化。从植入后早期胚胎、分离的内细胞团和滋养层细胞以及培养的胚胎干细胞制备了额外的样本。这些样本为鉴定细胞和组织特异性蛋白质以及表征胚胎早期细胞分化过程中伴随的蛋白质合成变化提供了手段。我们在此介绍小鼠胚胎数据库,并讨论其在哺乳动物胚胎发生研究中的潜在用途。