DUAL ADHESION MECHANISMS IN EMBRYONIC CHICK TISSUE

雏鸡胚胎组织中的双重粘附机制

• 批准号: 3315958
• 负责人: WILLIAM A THOMAS
• 金额: $ 6.36万
• 依托单位: WAKE FOREST UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-07-01 至 1988-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3315958
• 关键词: affinity chromatography; antibody; axon; calcium; carbohydrate structure; cell adhesion; cell aggregation; cell cell interaction; cell differentiation; chemical binding; electron microscopy; embryo /fetus cell /tissue; enzyme linked immunosorbent assay; fluorescence microscopy; fluorescent dye /probe; gel electrophoresis; image processing; immunoelectron microscopy; immunoprecipitation; molecular biology; monoclonal antibody; nonmammalian vertebrate embryology; retina; scintillation counter; superior colliculus; videotape /videodisc

Preferential cell associations arise in large part from differences in cellular adhesive properties with the result that regulation of those properties can in principle guide the organization of cells into anatomically "correct" structures. By corollary, errors in such regulation probably underlie at least in part the metastatic behavior of malignant cells and the morphogenetic aberrations leading to specific birth defects. A detailed knowledge of the molecular mechanisms generating cellular adhesive properties is thus essential to our understanding of diverse morphogenetic phenomena. In that context, this work proposes to investigate the role of the dual adhesion mechanisms described earlier in organizing the chick visual system. Recently generated monoclonals antibodies inhibitory of retinal Ca++ dependent (CD) aggregation will be used to identify and isolate components of the retinal CD adhesion mechanism. The molecules so identified will be characterized by molecular weight, pI and carbohydrate composition and their relationship to already identified CD molecules (ie., CaT, ligatin) determined. In exploring the regulatory potential of this mechanism, efforts will be made to elucidate the molecular basis for Ca++ dependent activity (cell-cell bonding) and protection from proteolysis. To complement this line of investigation experiments will also be performed to determine if molecules other than N-CAM contribute to retinal Ca++ independent (CI) adhesion. An array of monoclonal and monospecific polyclonal antibodies against CD and CI (non-CAM) adhesion molecules will then be used to 1) visualize the distribution of those molecules in the retina and other embryonic chick tissues, 2) to explore the structural relationship between functionally similar adhesion molecules from different tissues, 3) to investigate the role of CI and CD molecules in generating known adhesive gradients across the retina and tectum, and finally 4) to test the role of those in directing retino-tectal mapping. Cellular adhesiveness (rate, selectivity) will be tested using cell monolayer and aggregometer assays. Other work will be performed using various electrophoretic techniques, protein blotting, peptide mapping, affinity chromatography, immunochemistry and fluorescence microscopy.

优先细胞关联在很大程度上是由于细胞的差异而产生的 细胞粘附特性的结果是调节这些 原则上，特性可以引导细胞组织成 解剖学上“正确”的结构。 由此推论，此类监管中的错误 可能至少部分是恶性转移行为的基础 细胞和形态发生畸变导致特定的出生缺陷。 详细了解产生细胞的分子机制 因此，粘合性能对于我们理解不同的材料至关重要 形态发生现象。 在此背景下，这项工作建议 研究前面描述的双重粘附机制的作用 组织雏鸡视觉系统。 最近生成的单克隆抗体 抑制视网膜 Ca++ 依赖性 (CD) 聚集的抗体将 用于识别和分离视网膜 CD 粘连的成分 机制。 如此鉴定的分子将通过分子特征来表征 体重、PI 和碳水化合物组成及其与已 已确定的CD分子（即CaT、连接素）。 在探索中 该机制的监管潜力，将努力阐明 Ca++ 依赖性活性（细胞与细胞结合）的分子基础以及 防止蛋白水解。 为了补充这一调查线 还将进行实验以确定除 N-CAM 有助于视网膜 Ca++ 独立 (CI) 粘附。 一个数组 针对 CD 和 CI 的单克隆和单特异性多克隆抗体 （非 CAM）粘附分子将用于 1) 可视化 这些分子在视网膜和其他胚胎鸡中的分布 组织，2）探索功能之间的结构关系 来自不同组织的相似粘附分子，3）研究 CI 和 CD 分子在产生已知的粘合剂梯度中的作用 视网膜和顶盖，最后 4) 测试它们的作用 指导视网膜顶盖测绘。 细胞粘附性（速率、选择性） 将使用细胞单层和聚集计测定进行测试。 其他工作 将使用各种电泳技术、蛋白质 印迹、肽图谱、亲和层析、免疫化学和 荧光显微镜。