EGF AND ITS RECEPTORS IN EMBRYONIC DIFFERENTIATION

EGF 及其受体在胚胎分化中的作用

• 批准号: 2087729
• 负责人: EILEEN D ADAMSON
• 金额: $ 26.51万
• 依托单位: SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
• 依托单位国家: 美国
• 财政年份: 1980
• 资助国家: 美国
• 起止时间: 1980-07-01 至 1998-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2087729
• 关键词: SDS polyacrylamide gel electrophoresis; biological signal transduction; cell differentiation; cell growth regulation; embryo implantation; embryogenesis; embryonic stem cell; enzyme linked immunosorbent assay; epidermal growth factor; gel electrophoresis; gene expression; genetically modified animals; growth factor receptors; immunofluorescence technique; immunoprecipitation; laboratory mouse; neoplastic cell culture for noncancer research; nuclear runoff assay; polymerase chain reaction; regeneration; western blottings; wound healing

The overall aims are to understand the roles and activities of the epidermal growth factor receptor (EGFR) in developing embryos in vivo. Known effects of signalling after ligand binding to the EGFR range from changes in membrane motility, cell shape and epithelial polarity to the stimulation of proliferation or to increased differentiation. Over- expression of the EGFR gene in EC cells can lead to differentiation. In contrast, in non-embryonic cells, over-expression can lead to aberrant or transformed growth. Our understanding of how EGFR activities are modulated and how gene expression is regulated is still fragmentary. In the last grant period, we characterized the activities of a truncated, kinase- negative version of EGFR after transfection of the expression plasmid into Pl9 embryonal carcinoma (EC) cells. We showed that truncated EGFR acted as a dominant negative mutation, abrogating the activities of the normal EGF receptor. The result was inhibition of the process of differentiation to nervous tissues after induction by retinoic acid. The present proposal builds on and extends this approach. We propose to perturb the levels of EGFR in EC cells, embryonal stem (ES) cells and in embryos as an approach to understanding the roles of the EGFR in normal and aberrant growth. The specific aims are:- l. To study the roles of the EGFR in ES cells and in transgenic mice by introducing a dominant negative mutation of the receptor. 2. To study the roles of EGFRs in vitro and in vivo by over- expression of human EGFRs with and without accompanying transforming growth factor alpha (TGFalpha) expression in EC and ES cells and in transgenic mice. 3. To make initial studies of the roles and regulation of EGFRs in ovarian cells and tumors. 4. To determine some of the components in the EGFR signal transduction pathway in preimplantation embryos using antibodies and RT-PCR. A number of hypotheses will be tested. For instance, that autocrine-stimulated EGF-Rs provide a driving force to the continuation of differentiation and its irreversibility and stability such that when aberrant, tumors are initiated or stimulated to progress.

总体目标是了解组织的角色和活动 表皮生长因子受体（EGFR）在体内发育中的胚胎。 配体与 EGFR 结合后信号传导的已知影响范围包括 膜运动、细胞形状和上皮极性的变化 刺激增殖或增加分化。超过- EC细胞中EGFR基因的表达可以导致分化。在 相反，在非胚胎细胞中，过度表达可能导致异常或 转型增长。我们对 EGFR 活性如何调节的理解 基因表达的调控方式仍然不完整。在最后 在资助期间，我们表征了截短的激酶的活性 表达质粒转染后EGFR阴性版本 Pl9胚胎癌(EC)细胞。我们证明截短的 EGFR 充当 显性失活突变，消除正常 EGF 的活性 受体。结果是抑制了分化过程 视黄酸诱导后的神经组织。目前的建议 建立并扩展了这种方法。我们建议扰乱水平 EC 细胞、胚胎干 (ES) 细胞和胚胎中的 EGFR 作为一种方法 了解 EGFR 在正常和异常生长中的作用。这 具体目标是：- l．研究 EGFR 在 ES 细胞和 转基因小鼠通过引入显性失活突变 受体。 2. 通过过度研究EGFR在体外和体内的作用 人类 EGFR 的表达，有或没有伴随转化 EC 和 ES 细胞以及细胞中的生长因子 α (TGFα) 表达 转基因小鼠。 3. 初步研究其作用和调节 卵巢细胞和肿瘤中的 EGFR。 4. 确定一些组成部分 植入前胚胎中的 EGFR 信号转导通路 抗体和 RT-PCR。许多假设将得到检验。为了 例如，自分泌刺激的 EGF-R 为 分化的持续性及其不可逆性和稳定性 当异常时，肿瘤就会启动或刺激进展。