ROLE OF PDGF IN DEVELOPMENT

PDGF 在发育中的作用

• 批准号: 3329656
• 负责人: EILEEN D ADAMSON
• 金额: $ 17.31万
• 依托单位: SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-09-01 至 1995-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3329656
• 关键词: antisense nucleic acid; cell cycle; cell differentiation; early embryonic stage; embryo implantation; female; genetic manipulation; genetically modified animals; growth factor receptors; mammalian embryology; oligonucleotides; platelet derived growth factor; receptor expression

The overall aim is to determine what role PDGF plays in mammalian development. There is evidence that the PDGF-A gene is expressed in preimplantation mouse embryos while the B chain appears later in development, and in accord, the receptor that responds to the A chain (alpha receptor) appears before the beta receptor. Therefore, this study will focus on the early roles of the A chain and the alpha receptor on the development of the pre- and peri-implantation embryo. The specific aims are as follows: 1. to select those transgenic animals from those that have already been prepared that express excess amounts of PDGF or that have lower levels than the normal animal, and determine the effects of further inducing the transgene with zinc. Selected females will be induced with zinc in the drinking water during gestation to determine the correlations with developmental abnormalities. 2. To prepare antibodies to PDGFs and alpha and beta receptors for detection and quantitation of these polypeptides in transgenic mouse tissues. In addition, by making antibodies to the sites that are involved in ligand binding, we hope to select antisera that will block ligand binding to test the effect on embryo development in vitro. 3. To make a dominant negative construct for PDGF-R-alpha. A carboxyterminal truncated construct will be introduced in an expression vector to interfere with normal alpha receptor function and hence, determine the biological and developmental consequences of inactivated PDGF-receptors in vitro and in vivo. These studies will determine the roles of PDGF growth factors and their receptors in the processes of early cell division and cell differentiation. These animals will also be useful in studying the later consequences of abnormal PDGF and receptor levels in vascular disease and in tumor progression.

总体目标是确定 PDGF 在哺乳动物中发挥什么作用 发展。 有证据表明PDGF-A基因在 植入前小鼠胚胎，而 B 链出现较晚 发育，相应地，响应 A 链的受体 （α受体）出现在β受体之前。 因此，这 研究将集中于 A 链和 alpha 的早期作用 受体对植入前和植入前后胚胎发育的影响。 具体目的如下：1.筛选转基因动物 从那些已经准备好的表达过量的 PDGF 或水平低于正常动物，并确定 用锌进一步诱导转基因的效果。 已选择 女性在怀孕期间会通过饮用水中的锌来诱导 以确定与发育异常的相关性。 2. 到 制备 PDGF 以及 α 和 β 受体的抗体用于检测 以及转基因小鼠组织中这些多肽的定量。 在 此外，通过对参与配体的位点制备抗体 结合，我们希望选择能够阻止配体结合的抗血清 测试对体外胚胎发育的影响。 3. 打造主导地位 PDGF-R-α 的阴性构建体。 羧基末端被截短 将构建体引入表达载体中以干扰 正常的α受体功能，因此决定了生物和 失活的 PDGF 受体在体外和体内的发育后果 体内。 这些研究将确定 PDGF 生长因子的作用和 它们的受体在早期细胞分裂和细胞分裂过程中 差异化。 这些动物也将有助于研究 血管内 PDGF 和受体水平异常的后续后果 疾病和肿瘤进展。