FETAL LUNG BIOCHEMISTRY AND MICROANATOMY

胎肺生物化学和显微解剖学

• 批准号: 2216972
• 负责人: Stephen L. Young
• 金额: $ 15.48万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-05-01 至 1996-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2216972
• 关键词: antiserum; autoradiography; cell adhesion; cell cell interaction; cell cycle; cell migration; developmental genetics; embryo /fetus cell /tissue; extracellular matrix; fibroblasts; fibronectins; gene expression; growth /development; histogenesis; in situ hybridization; laminin; lung; lung alveolus; monoclonal antibody; nonmammalian vertebrate embryology; recombinant proteins; respiratory epithelium; tenascin; tissue /cell culture

Extracellular matrix (ECM) molecules such as fibronectin (FN) and laminin (LN) are restricted in their spatial and temporal expression. They have important influences upon cellular proliferation, differentiation and mobility. Antibodies to LN will inhibit key lung developmental processes like branching morphogenesis. A more recently described ECM molecule, tenascin (TN), also has a restricted spatio-temporal expression in mammalian lung. It may modulate the binding of cells to ECM substrates like FN and it can induce fibroblast metalloproteinase genes such as collagenase. My hypothesis is that cell-extracellular matrix interactions regulate pulmonary branching morphogenesis and alveolarization; I propose that TN is an ECM molecule which has a major role as a regulator of lung development and that inhibition of TN expression may be important in the response of the developing lung to postnatal injury. Using in vitro and in vivo methods we will investigate two key periods of lung development: Aim 1. Branching morphogenesis a) Determine the spatio-temporal expression of tenascin, including its alternatively spliced forms, in lung by morphologic measures at light and electron microscopic resolutions. b) Measure lung TN gene and protein expression. c) Test the functional importance of TN upon branching morphogenesis with polyclonal anti-TN antiserum and with monoclonal domain-specific anti-TN antibodies and recombinant expression proteins. d) Test whether interactions of tenascin with laminin and fibronectin modify adherence and spreading of fetal lung fibroblasts and epithelial cells on plastic substrate. Aim 2. Alveolarization: We will use autoradiography to measure cell kinetics of the lung parenchyma. Alveolar septal cellular proliferation will be compared to the spatio- temporal expression of lung tenascin. Completion of these specific aims will extend our current work on cellular kinetics and ECM structural molecules of growing lung. Previously identified milestones of lung epithelial development will be correlated with epithelial-mesenchymal interactions to be studied with this support. Understanding of mechanisms leading to bronchopulmonary dysplasia will be advanced.

细胞外基质 (ECM) 分子，例如纤连蛋白 (FN) 和层粘连蛋白 （LN）的空间和时间表达受到限制。 他们有 对细胞增殖、分化有重要影响 流动性。 LN 抗体将抑制关键的肺部发育过程 就像分支形态发生一样。 最近描述的 ECM 分子， tenascin (TN) 在以下细胞中也具有受限的时空表达： 哺乳动物的肺。 它可以调节细胞与 ECM 底物的结合 像 FN 一样，它可以诱导成纤维细胞金属蛋白酶基因，例如 胶原酶。 我的假设是细胞-细胞外基质 相互作用调节肺分支形态发生 肺泡化；我认为 TN 是一种 ECM 分子，其主要作用是 作为肺发育的调节剂和抑制 TN 的作用 表达可能对发育中的肺的反应很重要 产后损伤。 我们将使用体外和体内方法进行研究 肺发育的两个关键时期： 目标 1. 分支形态发生 a) 确定生腱蛋白的时空表达，包括其 选择性剪接形式，在光和光下通过形态学测量在肺中 电子显微镜分辨率。 b) 测量肺TN基因和蛋白 表达。 c) 测试 TN 在分支时的功能重要性 使用多克隆抗 TN 抗血清和单克隆抗血清进行形态发生 域特异性抗 TN 抗体和重组表达蛋白。 d) 测试生腱蛋白是否与层粘连蛋白和纤连蛋白相互作用 改变胎儿肺成纤维细胞和上皮细胞的粘附和扩散 塑料基质上的细胞。 目标 2. 肺泡化：我们将使用 放射自显影术测量肺实质的细胞动力学。 肺泡间隔细胞增殖将与空间进行比较 肺生腱蛋白的时间表达。 完成这些具体目标将扩展我们目前的工作 生长肺的细胞动力学和 ECM 结构分子。 先前确定的肺上皮发育里程碑将是 与待研究的上皮间质相互作用相关 这个支持。 了解导致支气管肺疾病的机制 发育不良将会进展。