FERTILITY OF DOMINANT NEGATIVE INHIBITION OF FGF IN UTERUS AND OVARIES

子宫和卵巢中 FGF 显性负抑制的生育力

• 批准号: 6318367
• 负责人: ROBERT D KOOS
• 金额: $ 13.96万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-04-01 至 2001-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6318367
• 关键词: cooperative study; fertility; fibroblast growth factor; genetically modified animals; growth factor receptors; histogenesis; hormone regulation /control mechanism; in situ hybridization; laboratory mouse; male reproductive system; ovary; polymerase chain reaction; receptor expression; reproductive development; uterus

Successful reproduction requires the cyclical growth differentiation, and death of cells in the ovary and uterus. These complex developmental processes are initiated by gonadotropic hormones from the pituitary and steroids from the ovary, but they are ultimately carried out by locally- produced growth factors. Many of the latter, including the fibroblast growth factors (FGFs), show distinct temporal and spatial patterns of expression in the ovary and uterus during the reproductive cycle or in response to hormonal stimulation. Physical and pharmacological ablation have defined many of the roles of the endocrine hormones in these processes, but those of the growth factors are less amenable to this approach: their roles can only be speculated at based on correlative expression studies. However, new transgenic techniques make it possible to precisely define the functions of FGFs during developmental processes. Studies in this laboratory show that cell-specific targeting of dominant- negative (dom-neg) FGF receptors (FGFRs) in reproductive tissues of transgenic mice is feasible. The goal of this study is to use dom-neg FGFRs, their expression targeted and controlled by cell-specific promoters, to inhibit the actions of FGFs at specific sites and times in the ovary and uterus. The study has four Specific Aims: Aim 1: To identify and localize FGFR subtypes in the mouse ovary and uterus. Aim 2:a) To produce mice that express dom-neg receptors for keratinocyte growth factor (KGF), an epithelial cell-specific FGF, and basic FGF (bFGF) in the epithelial cells of the endometrium using a 404 bp fragment of the uteroglobin promoter; and b) to evaluate uterine function in these mice. Aim 3:a) To produce mice that express dom-neg receptors for KGF and bFGF in the granulosa cells of the growing ovarian follicle using the -180 bp fragment of the Muellerian inhibiting substance promoter; and b) to evaluate ovarian function in these mice. Aim 4:a) To produce mice in which the expression of dom-neg FGFRs in the ovary and uterus can be temporarily controlled through tetracycline-responsive regulatory elements; and b) to evaluate the impact on fertility of switching on expression of dom-neg FGFRs in either the ovary or uterus at critical periods in the reproductive process (e.g., during follicular maturation or at the time of implantation). These studies will enhance our understanding of normal reproductive physiology as well as shed light on malfunctions in cell growth and differentiation in the ovary and uterus that lead to infertility, birth defects, and disease.

成功的繁殖需要周期性的生长分化，并且 卵巢和子宫中的细胞死亡。这些复杂的发展 这些过程是由促性腺激素启动的，来自脑下垂体和 卵巢中的类固醇，但它们最终是由局部- 产生了增长因素。后者中的许多，包括成纤维细胞 生长因子(FGFs)，表现出不同的时间和空间模式 在生殖周期的卵巢和子宫中的表达 对荷尔蒙刺激的反应。物理和药物消融 已经确定了内分泌荷尔蒙在这些 过程，但那些增长因素对此的适应程度较低 方法：只能根据相关性来推测他们的角色 表达研究。然而，新的转基因技术使我们有可能 准确定义FGFs在发育过程中的功能。 本实验室的研究表明，细胞特异性靶向显性- 大鼠生殖组织中(don-neg)成纤维细胞生长因子受体(FGFRs)的表达 转基因小鼠是可行的。这项研究的目的是使用DOM-Ng FGFRs，其表达受细胞特异性靶向和控制 启动子，以抑制FGFs在特定地点和时间的作用 卵巢和子宫。这项研究有四个具体目标：目标1：确定 并在小鼠卵巢和子宫中定位FGFR亚型。目标2：a) 产生表达角质形成细胞生长因子DOM-neg受体的小鼠 上皮细胞特异性成纤维细胞生长因子(KGF)和碱性成纤维细胞生长因子(BFGF)。 子宫内膜上皮细胞使用404个碱基片段 子宫球蛋白启动子；b)评估这些小鼠的子宫功能。 目的3：建立表达kgf和bfgg受体的小鼠。 在发育中的卵泡颗粒细胞中使用-180BP 米勒抑制物质启动子的片段；和b)至 评估这些小鼠的卵巢功能。目标4：a)生产出小鼠 DOM-neg FGFRs在卵巢和子宫中的表达可以是暂时的 通过四环素反应调控元件进行控制；以及b) 评价转阴对生育力的影响 FGFRs在卵巢或子宫中的表达 生殖过程(例如，在卵泡成熟期间或在 植入)。这些研究将加深我们对正常的理解 生殖生理学以及揭示细胞中的故障 卵巢和子宫的生长和分化导致 不孕不育、出生缺陷和疾病。