ANALYSIS OF CELL SURFACE DURING MAMMALIAN DEVELOPMENT

胚胎细胞表面的碳水化合物分子被认为 在发育过程中细胞与细胞的相互作用中发挥作用。 先前 其他人对小鼠胚胎的研究表明，细胞分化是 与细胞表面碳水化合物的变化有关。 这些措施包括重大改组，反映在核心预算的变化上。 合成和分子量概况，以及微妙的修改 糖链的末端残基。 提出的实验 这里将使用抗F9产生的单克隆抗体（抗IIC 3 胚胎癌细胞，其检测细胞表面碳水化合物 抗原（IIC 3）对小鼠胚胎的作用。 免疫荧光定位 实验表明，IIC 3的表达仅限于 与着床和胎盘形成有关的胚胎细胞。 使用此 单克隆抗体，我们将继续分析IIC 3抗原， 如下：（1）将IIC 3抗原免疫沉淀，并将其分子标记物与免疫沉淀物结合。 将通过柱色谱、SDS-聚丙烯酰胺凝胶 电泳和糖苷酶消化;（2）免疫沉淀 将来自F9细胞的材料与免疫沉淀的抗原进行比较 从胚胎来源，为差异或相似的分子 （3）IIC 3基因的发育表达分析 使用免疫荧光和免疫细胞化学，在体内和体外 （4）使用粘附测定分析其功能作用;以及 (5)分析了IIC 3表达的激素控制， 发育及其作为激素受体的潜在作用。 这些 实验将提供一种新的碳水化合物抗原的详细信息， 与老鼠的发育有关。 这些信息会帮助我们 理解细胞表面碳水化合物作为细胞生长的一部分的假定作用。 控制细胞分化的机制。

The carbohydrate molecules of the embryonic cell surface are thought to play a role in the cell to cell interactions during development. Previous work, by others, on mouse embryos suggests that cell differentiation is associated with changes in the carbohydrate displays on the cell surface. These include major reorganizations, as reflected by changes in core synthesis and molecular weight profiles, as well as subtle modifications of terminal residues of the carbohydrate chains. The experiments proposed here will employ a monoclonal antibody (anti-IIC3), produced against F9 embryonal carcinoma cells, which detects a cell surface carbohydrate antigen (IIC3) on mouse embryos. Immunofluorescent localization experiments have indicated that the expression of IIC3 is restricted to the cells of the embryo involved in implantation and placentation. Using this monoclonal antibody we will continue to analyze the IIC3 antigen as follows: (1) The IIC3 antigen will be immunoprecipitated and its molecular structure will be analyzed by column chromatography, SDS-polyacrylimide gel electrophoresis, and glycosidase digestion; (2) the immunoprecipitable material from F9 cells will be compared to the antigens immunoprecipitated from embryonic sources, for differences or similarities in molecular structure; (3) a continued analysis of the developmental expression of IIC3 using immunofluorescence and immunocytochemistry, on in vivo and in vitro systems; (4) an analysis of its functional role using adhesion assays; and (5) an analysis of the hormonal control of IIC3 expression during development and its potential role as a hormone receptor. These experiments will provide detailed information on a new carbohydrate antigen associated with mouse development. This information will help us understand the putative role of cell surface carbohydrates as part of the controlling mechanism of cellular differentiation.