CELL ADHESION & MOLECULE LOCALIZATION IN A MODEL SYSTEM

细胞粘附

• 批准号: 3439499
• 负责人: Steven Bernard Oppenheimer
• 金额: $ 7.14万
• 依托单位: CALIFORNIA STATE UNIVERSITY NORTHRIDGE
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-09-30 至 1988-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3439499
• 关键词: antibody formation; antibody specificity; biological models; cell adhesion; cell aggregation; cell cell interaction; cell growth regulation; contact inhibition; density gradient ultracentrifugation; disease /disorder model; electron microscopy; embryo /fetus cell /tissue; histogenesis; immunofluorescence technique; immunoglobulin G; metastasis; model design /development; molecular pathology; particle counter

The spread of cancer and embryonic morphogenesis are dependent upon changes in cellular adhesiveness. The molecular basis of cell-cell adhesion is poorly understood. This proposal presents a model system for studying cellular adhesion, the sea urchin embryo. Information gained from this system should be of general importance in improving our understanding of the nature of cell-cell interaction and possible reasons for the altered adhesiveness found in malignant cells. Molecules have been isolated in the calcium-magnesium-free sea water disaggregation supernatant of Strongylocentrotus purpuratus blastula embryos that promote cellular reaggregtion in a species-specific and embryonic stage-specific manner. By using fixed S. purpuratus blastula cells as adsorbants, a subset of molecules was isolated from the disaggregation supernatant. Rabbit antibody was made against this subset to examine its localization in sea urchin embryos via indirect immunofluorescence. Specific fluorescence was observed in a blastocoelar fibrillar matrix in S.purpuratus blastula embryos but not in embryos of other species or stages. This material may represent adhesive molecules that seep between cells and play important roles in the adhesive interactions of cells during morphogenesis. Non-specific antibodies will be removed from the polyclonal, polyspecific IgG by absorption with embryonic antigenic material other than S. purpuratus blastulae. The blastula specific antibody will be used in localization studies and Fab fragments of this antibody will be screened for ability to specifically interfere with cellular adhesion of S. purpuratus blastulae. Fibrillar blastocoelar matrices will be isolated from various species and stages of sea urchin embryos and screend for specific enhancement of sea urchin embryo cell aggregation. The final components of this study will involve screening antibodies against known extracellular molecules (and the molecules themselves) in localization and adhesion studies and sucrose and Ficoll gradient isolation of high molecular weight putative adhesion molecules in the disaggregation supernatant. This system presents a homogeneous synchronous, abundant model for identification of molecules that may function in specific cell adhesion.

癌症的扩散和胚胎形态发生取决于变化 在细胞粘附性方面。 细胞与细胞粘附的分子基础是 不太了解。 该提案提出了一个用于研究的模型系统 细胞粘附，海胆胚胎。 由此获得的信息 系统对于提高我们的理解应该具有普遍的重要性 细胞间相互作用的本质以及改变的可能原因 恶性细胞中发现的粘附性。 在不含钙镁的海水中分离出分子 紫圆甲藻囊胚的解聚上清液 胚胎在物种特异性和促进细胞重新聚集 胚胎阶段特定的方式。 通过使用固定的 S. purpuratus 囊胚 细胞作为吸附剂，从细胞中分离出一部分分子 分解上清液。 针对该子集制备了兔抗体 通过间接方法检查其在海胆胚胎中的定位 免疫荧光。 在囊胚腔中观察到特异性荧光 S.purpuratus 囊胚胚胎中存在纤维状基质，但在 其他物种或阶段。 这种材料可能代表粘合剂分子 渗透到细胞之间并在粘合剂中发挥重要作用 形态发生过程中细胞的相互作用。 非特异性抗体将从多克隆、多特异性抗体中去除 通过与 S 以外的胚胎抗原物质吸收而产生 IgG。 紫囊囊胚。 囊胚特异性抗体将用于 将筛选该抗体的定位研究和 Fab 片段 用于特异性干扰 S 细胞粘附的能力。 紫囊囊胚。 将分离纤维状囊胚腔基质 从不同物种和阶段的海胆胚胎中筛选 特异性增强海胆胚胎细胞聚集。 决赛 这项研究的组成部分将涉及针对已知的抗体进行筛选 细胞外分子（以及分子本身）的定位和 高浓度的粘附研究以及蔗糖和聚蔗糖梯度分离 解聚中假定的粘附分子的分子量 上清液。 该系统呈现出同质同步、丰富的 用于识别可能在特定细胞中起作用的分子的模型 附着力。