Murach K F, Frei M, Gerhäuser D, Illmensee K
Department of Animal Biology, University of Geneva, Switzerland.
J Cell Biochem. 1990 Sep;44(1):19-37. doi: 10.1002/jcb.240440103.
Mouse embryos of the NMRI strain between the 7th and 9th day of gestation were isolated from the uterus and dissected into the various tissue derivatives in order to investigate newly synthesized proteins during morphogenesis. The day 7 embryo was fragmented into trophoblast and ectoplacental cone, distal and proximal endoderm, extraembryonic and embryonic ectoderm. The day 8 and day 9 embryos were divided into trophoblast and placental anlage, yolk sac, amnion, and allantois, as well as cranial, central, and caudal embryonic tissue. The intact embryos were incubated in Dulbecco's minimum essential medium in the presence of 35S-methionine for 4 h, then dissected into the various fragments, and further processed for two-dimensional gel electrophoresis. Protein synthesis of the isolated tissue derivatives was analyzed and compared for the three developmental stages. Concerning the proteins with isoelectric points in the range of 4.5 to 8.0 and molecular weight ratio (M(r)) values between 20,000 and 200,000, we found several significant quantitative and qualitative differences in the various tissue fragments. In addition, we observed further quantitative and qualitative differences in protein synthesis during the postimplantation period investigated. We propose that the differences reflect some of the cell lineage- and developmental stage-specific changes in gene expression during early mammalian differentiation.
从妊娠第7天至第9天的NMRI品系小鼠胚胎中分离出子宫,并将其解剖成各种组织衍生物,以研究形态发生过程中新合成的蛋白质。将第7天的胚胎分割成滋养层和外胎盘锥、远端和近端内胚层、胚外和胚胎外胚层。将第8天和第9天的胚胎分为滋养层和胎盘原基、卵黄囊、羊膜和尿囊,以及头部、中央和尾部胚胎组织。将完整的胚胎在含有35S-甲硫氨酸的杜尔贝科改良伊格尔培养基中孵育4小时,然后解剖成各种片段,并进一步进行二维凝胶电泳处理。分析并比较了三个发育阶段分离的组织衍生物的蛋白质合成情况。对于等电点在4.5至8.0范围内且分子量比(M(r))值在20,000至200,000之间的蛋白质,我们在各种组织片段中发现了一些显著的定量和定性差异。此外,我们在研究的植入后时期观察到蛋白质合成的进一步定量和定性差异。我们认为这些差异反映了早期哺乳动物分化过程中基因表达的一些细胞谱系和发育阶段特异性变化。