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: 6108921
• 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: 1999
• 资助国家: 美国
• 起止时间: 1999-04-01 至 2000-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6108921
• 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 细胞在卵巢和子宫中的死亡。这些复杂的发展 过程是由垂体中的促性腺激素激素引发的， 卵巢的类固醇，但最终是由局部进行的 产生了生长因子。后者的许多，包括成纤维细胞 生长因子（FGF）显示出不同的时间和空间模式 在生殖周期或在卵巢和子宫中表达 对激素刺激的反应。物理和药理消融 已经定义了内分泌激素的许多角色 过程，但是增长因素的过程对此不太适合 方法：他们的角色只能基于相关性推测 表达研究。但是，新的转基因技术使得 精确定义了在发育过程中FGF的功能。 该实验室的研究表明，细胞特异性靶向显性靶向 负（DOM-neg）FGF受体（FGFR）的生殖组织 转基因小鼠是可行的。这项研究的目的是使用dom-neg FGFR，它们的表达因细胞特异性而针对和控制 发起人，抑制FGF在特定地点和时间的行为 卵巢和子宫。该研究具有四个具体目标：目标1：确定 并将FGFR亚型定位在小鼠卵巢和子宫中。目标2：a）到 产生表达角质形成细胞生长因子的dom-neg受体的小鼠 （kgf），上皮细胞特异性FGF和基本FGF（BFGF） 子宫内膜的上皮细胞使用404 bp的片段 子宫球蛋白启动子； b）评估这些小鼠的子宫功能。 目标3：a）产生表达kgf和bfgf的dom-neg受体的小鼠 在生长卵巢卵泡的颗粒细胞中，使用-180 bp 穆勒式抑制物质启动子的碎片；和b）到 评估这些小鼠的卵巢功能。目标4：a）生产小鼠 Dom-neg FGFR在卵巢和子宫中的表达可以暂时 通过四环素响应性调节元件进行控制；和b）到 评估对开关对DOM-NENG表达表达的影响的影响 在关键时期的卵巢或子宫中的FGFR 生殖过程（例如，在卵泡成熟期间或 植入）。这些研究将增强我们对正常的理解 生殖生理学以及阐明细胞中的故障 卵巢和子宫的生长和分化导致 不育，出生缺陷和疾病。