ANALYSIS OF CELL SURFACE DURING MAMMALIAN DEVELOPMENT

哺乳动物发育过程中细胞表面的分析

• 批准号: 3318815
• 负责人: BRUCE S BABIARZ
• 金额: $ 5.46万
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-09-01 至 1988-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3318815
• 关键词: antiantibody; autoradiography; carbohydrate structure; cell cell interaction; cell differentiation; cell type; electron microscopy; embryo /fetus antigen; embryo /fetus cell /tissue; gel electrophoresis; gel filtration chromatography; gene expression; histogenesis; hormone regulation /control mechanism; immunofluorescence technique; immunoprecipitation; mammalian embryology; membrane activity; membrane structure; microscopy; neoplastic cell culture for noncancer research; radiotracer; scintillation counter; surface antigens

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.

胚胎细胞表面的碳水化合物分子被认为是 在开发过程中，在细胞中发挥作用。 以前的 其他人在小鼠胚胎上的工作表明细胞分化是 与细胞表面上碳水化合物显示的变化有关。 这些包括重大的重组，如核心的变化所反映 合成和分子量谱，以及对 碳水化合物链的末端残基。 提出了实验 这里将使用针对F9产生的单克隆抗体（抗IIC3） 胚胎癌细胞，检测细胞表面碳水化合物 小鼠胚胎上的抗原（IIC3）。 免疫荧光定位 实验表明IIC3的表达仅限于 与植入和胎盘有关的胚胎细胞。 使用此 单克隆抗体我们将继续分析IIC3抗原 以下内容：（1）IIC3抗原将进行免疫沉淀，并将其分子免疫 结构将通过色谱柱色谱法，SDS-Polyacrylimide凝胶分析 电泳和糖苷酶消化； （2）免疫可抵消 将F9细胞的材料与抗原免疫沉淀的材料进行比较 来自胚胎来源，分子的差异或相似性 结构; （3）对IIC3的发育表达的持续分析 使用免疫荧光和免疫细胞化学，体内和体外 系统； （4）使用粘附测定法对其功能作用进行分析；和 （5）对IIC3表达的激素控制的分析 发育及其作为激素受体的潜在作用。 这些 实验将提供有关新碳水化合物抗原的详细信息 与小鼠发育有关。 这些信息将帮助我们 了解细胞表面碳水化合物的推定作用 细胞分化的控制机制。