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.

对控制羊水体积的控制的全面了解是 具有重大临床重要性，并促使了很多研究 其调节的机制。本申请提议阐明 血管内皮生长因子（VEGF）在促进的作用 羊膜和子叶乳清的血管生成和渗透性 血管并检查其在调节羊水体积调节中的作用 和构图。使用怀孕的绵羊作为动物模型， 将分子与生理方法相结合，五个具体目标是 建议的。 AIM 1将表征组织分布和丰度 胎儿，绒毛膜和胎盘子叶中VEGF蛋白和mRNA的 通过免疫组织化学，免疫测定，原位杂交，北印迹 分析和竞争性RT-PCR。假设是VEGF表达是 存在于与羊膜，绒毛膜和胎盘血相邻的细胞中 血管的水平高于其他细胞类型。 AIM 2意志 确定VEGF受体mRNA的细胞分布和表达 在胎膜和胎盘中。假设是VEGF受体是 在这些组织的血管内皮细胞中表达。 目标3意志 研究胎儿膜中VEGF基因表达的调节和 缺氧和胎儿食管结扎的胎盘。假设是 胎儿缺氧或食道阻塞会诱导VEGF基因表达。目标4 将确定VEGF在胎儿膜中的生物学功能和 胎盘。 血管血管生成将通过形态计量计算 分析和通过渗透率估算的血管通透性 放射性白蛋白，钠和尿素。假设是VEGF具有 这些组织中的血管生成和渗透率。目标5意志 利用带有主要阴性突变体的转基因小鼠模型 使受体无效的VEGF受体。假设 是否缺乏功能性VEGF受体会导致异常 由于减少而怀孕期间的羊水体积和成分 膜中的微固定密度和渗透性。提议 研究将提供有关调节的重要信息 微血管网络灌注胎儿膜和胎儿表面 胎盘。这些信息将大大改善 了解怀孕期间羊水体积调节。