ROLE OF CELLULAR RECOGNITION IN CARDIAC MORPHOGENESIS

目的是研究特定大分子的作用 参与细胞的识别（细胞-细胞）， 细胞外基质成分（cell-ECM）在心脏形态发生中的作用。 将进行研究以：1）分离和表征细胞 参与细胞-细胞和细胞-ECM重组的表面大分子; 2） 开发针对这些分离的多克隆和单克隆抗体 3）进行生物测定以确定这些大分子的作用; 在心脏发育的特定时间的大分子; 4）分析 这些细胞表面成分和细胞骨架之间的联系 5）检测这些成分的表达 在特定的形态发生时期使用cDNA技术和图像 增强分析 心脏形成的基础是 在所有发育阶段的特定细胞-细胞和细胞-ECM相互作用 从胚胎到成人。 这三者之间的复杂互动 心脏的主要细胞类型（肌细胞、成纤维细胞和内皮细胞） 细胞）将被检查。 最初，将进行细胞粘附试验 胎儿、新生儿和成人肌细胞、成纤维细胞和内皮细胞 确定在识别的异同 用于纯化ECM组分如纤连蛋白、层粘连蛋白和 各种胶原蛋白。 细胞-细胞粘附也将用 这些相同的细胞类型。 使用免疫学方法，抗体将 针对纯化的提取物制备至细胞表面，并用于：1） 抑制细胞-细胞和/或细胞-ECM相互作用，和2）纯化 和特定大分子的鉴定。 产生的抗体 将在不同的时间检查细胞表面成分， 利用重组cDNA技术和荧光技术 显微镜 细胞分选在心脏形成中的作用将 在开发出3种特异性细胞表面探针后进行检查 主要细胞类型 这些细胞表面的特异性相互作用 与心脏形态发生有关的细胞骨架成分将是 使用针对细胞表面标志物的抗体和各种 细胞骨架的组成部分。 通过这些调查， 细胞-细胞和细胞-ECM受体，我们将试图进一步阐明 心脏形态发生的复杂生化控制。

The objective will be to investigate the role of specific macromolecules involved in the cellular recoginition (cell-cell) and in the recognition of the extracellular matrix components (cell-ECM) in cardiac morphogenesis. Investigations will be conducted to: 1) isolate and characterize cell surface macromolecules involved in cell-cell and cell-ECM recongintion; 2) develop both polyclonal and monoclonal antibodies to these isolated macromolecules; 3) conduct bioassays to determine the role of these macromolecules at specific times of development of the heart; 4) analyse the association between these cell surface components and the cytoskeleton of developing myocytes; and 5) examine the expression of these components at specific periods of morphogenesis using cDNA technology and image enhancement analysis. Fundemental to the formation of the heart are the specific cell-cell and cell-ECM interactions at all stages of development from the embryo to the adult. These complex interactions between the three principal cell types of the heart (myocytes, fibroblasts and endothelial cells) will be examined. Initially, cell adhesion assays will be conducted with fetal, neonate and adult myocytes, fibroblasts and endothelial cells to determine the similarities and differences in the recognition of the cell types for purifies ECM components such as fibronectin, laminin and various types of collagens. Cell-cell adhesion will also be assayed with these same cell types. Using an immunological approach, antibodies will be prepared against purified extracts to the cell surface and used for: 1) inhibition of cell-cell and/or cell-ECM interactions, and 2) purification and identification of the specific macromolecule(s). Antibodies produced against cell surface components will be examined at different times of development using recombinant cDNA technology and by fluorescent microscopy. The role of cell sorting in the formation of of the heart will be examined after the development of specific cell surface probes of the 3 principal cell types. The specific interaction of these cell surface components with cytoskeleton in relation to cardiac morphogenesis will be investigated using antibodies against the cell surface markers and various components of the cytoskeleton. Through these investigations on the role of cell-cell and cell-ECM receptors we will attempt to further elucidate the complex biochemical controls of cardiac morphogenesis.