CELL--ECM INTERACTIONS IN MYOCARDIAL FORM AND FUNCTION

心肌形态和功能中的细胞-ECM 相互作用

• 批准号: 3362274
• 负责人: MICHAEL SOLURSH
• 金额: $ 8.35万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-04-01 至 1994-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3362274
• 关键词: cell cell interaction; cell differentiation; cell migration; collagen; complementary DNA; congenital heart disorder; embryo /fetus culture; enzyme linked immunosorbent assay; extracellular matrix; heart; heart function; histogenesis; immunochemistry; immunofluorescence technique; in situ hybridization; intermediate filaments; laboratory mouse; laboratory rat; mammalian embryology; mesenchyme; messenger RNA; myocardium; nucleic acid probes

Cell:extracellular matrix (ECM) interactions play a critical role in early cardiac morphogenesis but what is known has been derived from avian models. Elucidation of these interactions in a mammalian model is essential if we are to begin to investigate the mechanisms of congenital heart disease. We propose to delineate, in the mammalian embryo (mouse and rat), the distribution and developmental appearance of the major known components of the extracellular matrix (collagen types I, III, IV, hyaluronic acid, laminin, fibronectin, basement membrane heparan sulfate proteoglycan) as they relate to precardiac cell migration an early cardiac morphogenesis. A) We will employ whole embryo mounts in combination with serial sections labelled with antibodies specific to each component of the ECM to define the developmental appearance and three dimensional distribution of these molecules in relation to the precardiac mesoderm. B) We will then utilize in situ hybridization with cDNA probes for the mRNA's of relevant ECM components (Collagen I,III,IV, fibronectin, laminin) to localize the temporal and geographical distribution of cells capable of influencing the precardiac cell population during the period of early cardiac morphogenesis. In addition, we will examine immunohistochemical markers for precardiac mesenchyme that will distinguish these cells from the surrounding lateral plate mesoderm at progressive stages in their migration and differentiation. Exploiting the unique character of these cells as migrating myoblasts, we will use monoclonal antibodies directed against embryonic myosins, desmin intermediate filaments, and myoblast specific cell surface epitopes to: a) identify the precardiac mesenchyme as a subpopulation of the lateral plate mesoderm, b) define the precise domain of precardiac cell migration, and c) determine the sequence in which these epitopes are expressed during precardiac cell differentiation. Additional monoclonal antibodies directed against precardiac mesenchyme will be developed, if needed, in order to recognize earlier stages of cardiac morphogenesis. Finally, we will investigate, in situ, the cell- ECM interactions involved in precardiac cell migration, differentiation, and early contractile function in the mammalian embryo. We will culture whole embryos during the period from gastrulation through cardiac looping in the presence of inhibitors of normal cell:ECM interaction. Embryos will be evaluated for resulting structural alteration and the effects of treatment will be quantified through sequential motion analysis of myocardial contractile function. This should provide evaluation of the relative contributions of the ECM components to normal structural and functional development.

细胞：细胞外基质 (ECM) 相互作用发挥着关键作用 在早期心脏形态发生中，但已知的已经衍生出来 来自鸟类模型。 阐明这些相互作用 如果我们要开始研究哺乳动物模型是必不可少的 先天性心脏病的发病机制。 我们建议划定， 在哺乳动物胚胎（小鼠和大鼠）中，分布和 主要已知成分的发育外观 细胞外基质（I、III、IV 型胶原蛋白、透明质酸、 层粘连蛋白、纤连蛋白、基底膜硫酸乙酰肝素 蛋白多糖），因为它们与心前细胞迁移和早期 心脏形态发生。 A) 我们将采用全胚胎封固 与带有特异性抗体标记的连续切片组合 ECM 的每个组成部分来定义发育外观 以及这些分子的三维分布关系 至心前中胚层。 B) 然后我们将就地利用 与相关 ECM mRNA 的 cDNA 探针杂交 成分（胶原蛋白 I、III、IV、纤连蛋白、层粘连蛋白）来定位 细胞的时间和地理分布 影响心前细胞群 早期心脏形态发生。 此外，我们将检查 心前质间质的免疫组织化学标记物 将这些细胞与周围的侧板中胚层区分开来 处于迁移和分化的渐进阶段。 利用这些细胞迁移的独特特性 成肌细胞，我们将使用针对成肌细胞的单克隆抗体 胚胎肌球蛋白、结蛋白中间丝和成肌细胞 特定的细胞表面表位，用于： a) 识别心前病变 间充质作为侧板中胚层的一个亚群，b) 定义心前细胞迁移的精确域，以及 c) 确定这些表位在过程中表达的序列 心前细胞分化。 其他单克隆抗体 将开发针对心前质间质的药物，如果 需要，以便识别心脏病的早期阶段 形态发生。 最后，我们将原位研究细胞- ECM 相互作用参与心前细胞迁移， 哺乳动物的分化和早期收缩功能 胚胎。 我们将在这段时间内培养整个胚胎 在存在抑制剂的情况下通过心脏循环进行原肠胚形成 正常细胞：ECM 相互作用。 胚胎将被评估 由此产生的结构改变和治疗效果将 通过心肌的连续运动分析来量化 收缩功能。 这应该提供对 ECM 成分对正常结构的相对贡献 和功能开发。