REGULATION OF VEGF EXPRESSION IN PLACENTA AND MEMBRANES

胎盘和胎膜中 VEGF 表达的调节

• 批准号: 2403516
• 负责人: CECILIA CHEUNG
• 金额: $ 18.35万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-09-01 至 1999-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2403516
• 关键词: amniotic fluid; angiogenesis; blood vessels; chorioallantoic membrane; embryo /fetus membrane; gene expression; growth factor; growth factor receptors; hormone regulation /control mechanism; immunocytochemistry; immunologic assay /test; in situ hybridization; membrane permeability; messenger RNA; northern blottings; placenta; polymerase chain reaction; sheep; vascular endothelial growth factors

A comprehensive understanding of the control of amniotic fluid volume is of major clinical importance and has prompted much research into the mechanism of its regulation. The present application proposes to elucidate the role of vascular endothelial growth factor (VEGF) in promoting angiogenesis and permeability of amniotic, chorionic and cotyledonary blood vessels and examine its role in regulation of amniotic fluid volume and composition. Using the pregnant sheep as the animal model, and combining molecular with physiological approaches, five specific aims are proposed. Aim 1 will characterize the tissue distribution and abundance of VEGF protein and mRNA in fetal amnion, chorion and placental cotyledons by immunohistochemistry, immunoassay, in situ hybridization, Northern blot analysis and competitive RT-PCR. The hypothesis is that VEGF expression is present in cells adjacent to the amniotic, chorionic and placental blood vessels at levels higher than that in other cell types. Aim 2 will determine the cellular distribution and expression of VEGF receptor mRNA in fetal membranes and placenta. The hypothesis is that VEGF receptors are expressed in vascular endothelial cells of these tissues. Aim 3 will investigate the regulation of VEGF gene expression in fetal membranes and placenta by hypoxia and fetal esophageal ligation. The hypothesis is that fetal hypoxia or esophageal occlusion induces VEGF gene expression. Aim 4 will determine the biological function of VEGF in fetal membranes and placenta. Vascular angiogenesis will be evaluated by morphometric analysis, and vascular permeability estimated by permeability to radioactive albumin, sodium and urea. The hypothesis is that VEGF has both angiogenic and permeability properties in these tissues. Aim 5 will utilize a transgenic mouse model carrying a dominant-negative mutant of the VEGF receptor which renders the receptor ineffective. The hypothesis is that the lack of a functional VEGF receptor results in abnormal amniotic fluid volume and composition during pregnancy due to reduction in microvessel density and permeability in the membranes. The proposed studies will provide important information on the regulation of the microvascular network which perfuses the fetal membranes and fetal surface of the placenta. This information will significantly improve the understanding of amniotic fluid volume regulation during pregnancy.

全面了解羊水量的控制是 具有重要的临床意义，并引发了许多研究 其调节机制。本申请旨在阐明 血管内皮生长因子（VEGF）在促进血管生成中的作用 羊膜、绒毛膜和子叶的血管生成和通透性 血管并检查其在调节羊水量中的作用 和组成。以怀孕羊为动物模型， 结合分子与生理学方法，五个具体目标是 建议的。 目标 1 将表征组织分布和丰度 胎儿羊膜、绒毛膜和胎盘子叶中 VEGF 蛋白和 mRNA 的变化 通过免疫组织化学、免疫分析、原位杂交、Northern印迹 分析和竞争性 RT-PCR。假设 VEGF 表达是 存在于羊膜血、绒毛膜血和胎盘血附近的细胞中 血管中的水平高于其他细胞类型。目标2将 测定 VEGF 受体 mRNA 的细胞分布和表达 存在于胎膜和胎盘中。假设 VEGF 受体是 在这些组织的血管内皮细胞中表达。 目标3将 研究胎膜中 VEGF 基因表达的调控 胎盘经缺氧并结扎胎儿食管。假设是 胎儿缺氧或食管阻塞会诱导VEGF基因表达。目标 4 将决定胎膜中 VEGF 的生物学功能 胎盘。 血管生成将通过形态测量进行评估 分析，并通过渗透性估计血管渗透性 放射性白蛋白、钠和尿素。假设 VEGF 具有两者 这些组织中的血管生成和渗透性特性。目标5将 利用携带显性失活突变体的转基因小鼠模型 VEGF 受体，使受体无效。假设 缺乏功能性 VEGF 受体会导致异常 由于怀孕期间羊水量和成分减少 膜中的微血管密度和渗透性。拟议的 研究将提供有关监管的重要信息 灌注胎膜和胎儿表面的微血管网络 胎盘。该信息将显着改善 了解怀孕期间羊水量的调节。