EXTRACELLULAR MATRIX INTERACTIONS IN CARDIAC MORPHOGENESIS

心脏形态发生中的细胞外基质相互作用

• 批准号: 6109979
• 负责人: MICHAEL SOLURSH
• 金额: $ 17.73万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-01-30 至 2000-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6109979
• 关键词: animal genetic material tag; blocking antibody; cardiovascular disorder epidemiology; cell cell interaction; cell differentiation; cell migration; chick embryo; collagen; congenital heart disorder; early embryonic stage; elastin; extracellular matrix; fibronectins; heart cell; heart function; heart valves; histogenesis; homeobox genes; immunocytochemistry; in situ hybridization; laboratory mouse; laboratory rat; laminin; molecular genetics; myosins; tissue /cell culture

A large proportion of congenital heart defects are seen in the valves and membranous septa of the heart, which are in part derived from the endocardial cushions. The morphogenesis of the endocardial cushions is a complex process, involving a sequence of basic developmental events. These include cell migration, proliferation, and differentiation. It is the goal of this project to define this developmental sequence in greater detail and to understand the role of specific genes in the process. The major focus is on components of the extracellular matrix. The following questions will be addressed. l) What is the role of the homeobox gene, Hox 7, in the initiation of cushion morphogenesis? This will be addressed by examining the effects of anti-sense deoxyoligonucleotides on cushion morphogenesis and on the expression of extracellular matrix components in heart explant cultures and in ovo. 2) What extracellular matrix molecules are expressed during cushion formation? This will be addressed by immunohistochemistry with a battery of antibodies available in the laboratory and by in situ hybridization in some cases of special interest. 3) What is the developmental effect of blocking the expression of specific extracellular matrix components on cushion morphogenesis? This will be carried out with blocking antibodies, where available, or anti-sense deoxyoligonucleotides. In particular we will focus on hyaluronan and its binding proteins (CD-44 and PG-M), collagen II, and a component of the cardiac jelly (1E12) which has been identified recently in this laboratory. 4) The role of other genes in cushion morphogenesis will be studied subsequently, as they become characterized. 5) What is the contribution of the cushion cells to the tissues and extracellular matrix of the mature valves and septa of the heart? This question will be addressed by immunohistological analysis after in situ labeling of cushion cells with replication-defective, lac-Z labeled retrovirus, followed by the reincubation of the windowed, injected chicken eggs.

很大一部分先天性心脏病见于瓣膜和 心脏的膜隔膜，部分源自 心内膜垫。心内膜垫的形态发生是 复杂的过程，涉及一系列基本的发展事件。这些 包括细胞迁移、增殖和分化。这是目标 该项目的目的是更详细地定义该开发顺序 并了解特定基因在此过程中的作用。主要 重点是细胞外基质的成分。下列 问题将得到解决。 l) 同源盒基因Hox的作用是什么 7、缓冲垫形态发生的启动？这将由 检查反义脱氧寡核苷酸对缓冲的影响 形态发生和细胞外基质成分的表达 心脏外植体培养和卵内培养。 2）细胞外基质分子有哪些 在缓冲层形成过程中表达？这将由 免疫组织化学与一组可用的抗体 实验室和在某些特别感兴趣的情况下通过原位杂交。 3) 阻断特定基因的表达对发育有何影响？ 细胞外基质成分对缓冲垫形态发生的影响？这将是 使用封闭抗体（如果有）或反义抗体进行 脱氧寡核苷酸。我们将特别关注透明质酸及其 结合蛋白（CD-44 和 PG-M）、II 型胶原蛋白以及 最近在本报告中鉴定出心脏胶 (1E12) 实验室。 4）其他基因在垫形态发生中的作用将是 随着它们的特征化，随后进行了研究。 5) 什么是 垫细胞对组织和细胞外基质的贡献 心脏成熟的瓣膜和隔膜？这个问题将是 垫原位标记后通过免疫组织学分析解决 具有复制缺陷、lac-Z 标记逆转录病毒的细胞，然后 重新孵化带窗口的注射鸡蛋。