MAPPING SIGNAL TRANSDUCTION IN VASCULAR ENDOTHELIUM

绘制血管内皮信号转导图

• 批准号: 6152698
• 负责人: THOMAS N SATO
• 金额: $ 33.7万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-30 至 2004-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6152698
• 关键词: biological models; biological signal transduction; biotechnology; cardiovascular function; cell line; electroporation; embryonic stem cell; flow cytometry; genetically modified animals; high throughput technology; hypoxia; laboratory mouse; method development; model design /development; molecular biology information system; polymerase chain reaction; protein kinase; southern blotting; tissue /cell preparation; transfection; transforming growth factors; vascular endothelium

DESCRIPTION (Adapted from the applicant's abstract) My broad and long-term objective is to develop murine indicator lines that allow us to selectively determine alterations of signal transduction in vascular endothelium in an inexpensive, high-throughput format. Vascular endothelium plays a number of critical roles in normal development and homeostasis of the cardiovascular system and in the pathogenesis of many human cardiovascular diseases. Vascular endothelium senses a variety of chemical and physical changes occurring in its microenvironment. Aberrant regulation of this sensing mechanism, called signal transduction, in vascular endothelium often leads to dysfunction of this cell type, and is manifested as many forms of diseases. The mouse is an excellent animal model to study human diseases. In past years, numerous mutant mice have been generated by either targeted or random mutagenesis, and more mutants will be generated at an accelerated rate. Therefore, development of inexpensive high-throughput phenotypic screening methods will be essential for us to catch up with the rate at which mutant mice are generated. In this proposal, I focus on the development of a method that allows us to determine biochemical alterations of specific signal transduction pathways selectively in vascular endothelial cells. This goal will be accomplished with the following six specific aims: (1) To develop mouse indicator lines that allow the measurement of alterations of specific signal transduction pathways selectively in vascular endothelial cells. (2) to establish temporal and spatial maps of activation and inactivation of each of the signal transduction pathway in vascular endothelium by using the indicator lines. (3) To establish a biochemical assay in a high-throughput format by using tissues from the indicator lines. (4) To generate these indicators in various genetic backgrounds to allow immediate crossing to the existing and future mutant mice. (5) To generate embryonic stem cell lines from the indicator lines. (6) To establish a web site to streamline the dissemination oft he reagents, protocols, and information established through this project. Availability of our high-throughput screening method should lead to our further understanding development and physiological function of the cardiovascular system. Furthermore, the knowledge gained through the screening may result in the emergence of new concepts related to the regulation of cardiovascular development and function. (End of Abstract.)

描述（根据申请人的摘要改编） 我广泛的长期目标是开发鼠指标线 允许我们选择性确定信号转导的改变 廉价，高通量格式的血管内皮。 血管 内皮在正常发育中起许多关键作用， 心血管系统的稳态和许多人的发病机理 心血管疾病。 血管内皮感应多种化学物质 以及其微环境中发生的身体变化。 异常法规 在血管内皮中，这种传感机制，称为信号转导 通常会导致这种细胞类型的功能障碍，并且表现出多种形式 疾病。 小鼠是研究人类疾病的出色动物模型。 在过去的几年中，靶向或 随机诱变和更多的突变体将以加速速率产生。 因此，开发廉价的高通量表型筛选 方法对于我们赶上突变体的速率至关重要 小鼠是生成的。 在此提案中，我专注于一种方法的发展 这使我们能够确定特定信号的生化改变 在血管内皮细胞中选择性转导途径。 这个目标 将通过以下六个具体目标来完成：（1）开发 允许测量特定变化的鼠标指示线 信号转导途径在血管内皮细胞中有选择地。 （2） 建立每个激活和灭活的时间和空间图 通过使用该信号转导途径在血管内皮中使用 指示线。 （3）在高通量中建立生化测定 通过使用指示线的组织格式。 （4）生成这些 各种遗传背景的指标允许立即交叉 现有的和未来的突变小鼠。 （5）生成胚胎干细胞系 从指示线。 （6）建立一个网站以简化 通过的试剂，协议和信息通过 这个项目。 我们的高通量筛选方法的可用性应 导致我们进一步了解发展的发展和生理功能 心血管系统。 此外，通过 筛选可能会导致出现与 心血管发育和功能的调节。 （摘要结束。）