CELL-MATRIX INTERACTIONS DURING MORPHOGENESIS

形态发生过程中的细胞-基质相互作用

• 批准号: 3322262
• 负责人: Stuart A. Newman
• 金额: $ 11.7万
• 依托单位: NEW YORK MEDICAL COLLEGE
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-04-01 至 1990-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3322262
• 关键词: cartilage development; cell cell interaction; cell differentiation; cell membrane; chick embryo; cinematography; extracellular matrix; fibronectins; histogenesis; limbs; microscopy; nonmammalian vertebrate embryology; tissue /cell culture

The mechanisms of normal and abnormal tissue morphogenesis will be studied in cell cultures, chicken embryos and in a model collagen gel system. Polystyrene latex beads will be used as probes of cell-extracellular matrix interactions during morphogenesis of limb tissues in these systems. Heparin-coated beads have been found previously to be translocated into developing mesenchymal aggregates in high density cultures of chick limb precartilage cells. They are also subject to translocating forces generated in type I collagen gels in which fibronectin is nonuniformly distributed. Using beads coated with various cell surface macromolecules and monoclonal antibodies directed against specific domains of fibronectin, the cell surface and extracellular matrix determinants of translocation in the culture and gel systems will be studied. Cell aggregation and chondrogenesis will be assayed in high density cultures containing anti-fibronectin antibodies to analyze the matrix determinants of precartilage cell aggregation and differentiation. Beads with various surface coatings will be micro-injected into the somites and limb buds of chick embryos to assay selective translocation into chondrogenic or myogenic primordia of the limbs. Elvax resin containing anti-fibronectin monoclonal antibodies will be implanted in developing limbs with or without bead injections to assay the fibronectin domain dependence of potential bead translocation during limb development, as well as the role of specific fibronectin domains in chondrogenic and myogenic cell translocation and morphogenesis. The relevance of the collagen gel model to cell-matrix interactions in living tissues will be studied with additions of hyaluronic acid, chondroitin sulfate and type III collagen to gels containing varying amounts of type I collagen. In addition, the local organization of collagen fibrils during cell and bead translocation in collagen gels will be examined using polarizing microscopy, and the magnitude of the translocational force will be determined by measurement of the magnetic field strength necessary to oppose translocation of heparin-coated iron oxide spheres. These studies will provide insight into the molecular mechanisms of morphogenetic interaction during normal and abnormal embryonic development.

将研究正常和异常组织形态发生的机制 在细胞培养物，鸡胚胎和模型胶原蛋白凝胶系统中。 聚苯乙烯乳胶珠将用作细胞 - 细胞基质的探针 这些系统中肢体组织形态发生过程中的相互作用。 以前发现肝素涂覆的珠被转移到 在雏鸡肢体的高密度培养物中发展间充质聚集体 前后细胞。 他们也受到易位力量 在纤连蛋白不均匀的I型胶原蛋白凝胶中产生 分布式。 使用涂有各种细胞表面大分子的珠子 和针对纤连蛋白特定结构域的单克隆抗体， 易位的细胞表面和细胞外基质决定因素 将研究培养和凝胶系统。 细胞聚集和 软骨发生将在含有的高密度培养物中测定 抗纤维蛋白抗体分析基质决定因素 前电池的聚集和分化。 珠子有多种 表面涂层将被微型注射到各个节和四肢芽中 雏鸡胚胎选择性转运为软骨或 四肢的肌原始。 含有抗纤维蛋白的猫科树脂 单克隆抗体将植入有或没有的四肢发育 注射珠以测定纤连蛋白域的依赖性 肢体发育过程中的珠易位以及特定的作用 软骨和肌原性细胞易位中的纤连蛋白结构域以及 形态发生。 胶原蛋白凝胶模型与细胞矩阵的相关性 将研究透明质酸的生活组织中的相互作用 酸，硫酸软骨素和III型胶原蛋白到含有变化的凝胶 I型胶原蛋白的数量。 此外，本地组织 胶原蛋白凝胶中的胶原蛋白原纤维和珠子易位将 使用偏振显微镜检查和 易位力将通过测量磁性来确定 反对肝素涂层铁的易位所需的野外强度 氧化物球。 这些研究将提供有关分子的洞察力 正常和异常的形态发生相互作用的机制 胚胎发展。