INSULIN AND IGF-1 SIGNALING IN OVARIAN CELLS

卵巢细胞中的胰岛素和 IGF-1 信号传导

• 批准号: 6233079
• 负责人: DAVID W SCHOMBERG
• 金额: $ 7.7万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-04-01 至 2003-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6233079
• 关键词: apoptosis; biological signal transduction; biomarker; cell cycle; egg /ovum; female; follicle stimulating hormone; gene targeting; genetically modified animals; gonadotropins; granulosa cell; growth factor receptors; hormone receptor; hormone regulation /control mechanism; immature animal; immunocytochemistry; insulin; insulinlike growth factor; laboratory mouse; ovary; polymerase chain reaction; tissue /cell culture; western blottings

DESCRIPTION (Adapted from applicant's description): Our preliminary studies of ovaries from transgenic mice with a knockout of the insulin receptor substrate-1 gene (IRS-1 KO) showed entrapment of the oocyte in luteal tissue and that the number of animals ovulating was about one-third that of the wild- type (WT) control group. The IRS-1 KO ovaries also exhibited a rare phenotype, polyovular (bi-oocyte) follicles. These findings imply a relationship between compromised insulin signaling and phenotypic change in ovarian function which have not been previously recognized. Accordingly, the overall study objective is to validate and extend these findings to establish the role of insulin signaling pathway components at the ovarian level in vivo. With this background, detailed studies of how these effectors help execute the insulin- and Follicle-stimulating hormone-FSH) regulated end point alternatives of cell survival, apoptosis, or mitosis will proceed in an expanded application. The ovaries of WT and IRS-1 KO mice will be compared in various aspects in the following specific aims: 1) to evaluate oocyte numbers and biochemical markers in neonatal animals and the ovulatory response to exogenous gonadotropins in prepubertal animals, and 2) to assess levels and/or activity of early effectors in the insulin signaling pathway (IRS-1, IRS-2, P13-K, Akt) during gonadotropin-regulated follicular cell mitosis and apoptosis. Specific Aim 3 will extend the in vivo studies to in vitro analysis in cultured porcine granulosa cells (pGCs) to: a) examine possible connections between the FSH receptor and insulin/IGF-1 receptor signaling pathways, and b) develop improved methods to detect BAD (Bcl-2-associated death promoter), a critical regulator of cell fate which can be phosphorylated by Akt. The results obtained may also contribute information basic to a more complete understanding of an important clinical entity, polycystic ovarian syndrome (PCOS), which is characterized in part by accelerated follicle atresia (apoptosis), insulin resistance, and virilism.

描述（改编自申请人的描述）：我们的初步研究 胰岛素受体敲除的转基因小鼠的卵巢 底物-1基因（IRS-1 KO）显示卵母细胞在黄体组织中被截留 排卵的动物数量大约是野生动物的三分之一 WT型对照组。IRS-1 KO卵巢也表现出罕见的表型， 多卵（双卵母细胞）卵泡。这些发现暗示了 胰岛素信号传导受损和卵巢功能表型改变， 以前没有得到承认。 因此，总体研究目标 是验证和扩展这些发现，以确定胰岛素的作用， 在体内卵巢水平的信号通路组分。与此 背景，这些效应器如何帮助执行胰岛素的详细研究， 和卵泡刺激素-FSH）调节的细胞终点替代物 存活、凋亡或有丝分裂将在扩展的应用中进行。 的 WT和IRS-1 KO小鼠的卵巢将在各个方面进行比较， 以下具体目的：1）评估卵母细胞数量和生化标志物 在新生动物和排卵反应的外源性促性腺激素， 青春期前动物，和2）评估早期的水平和/或活动 胰岛素信号通路中的效应物（IRS-1、IRS-2、P13-K、Akt） 促性腺激素调节的卵泡细胞有丝分裂和凋亡。 具体目标3 将体内研究扩展到培养猪的体外分析 颗粒细胞（pGC）：a）检查FSH与卵泡刺激素（FSH）之间的可能联系， 受体和胰岛素/IGF-1受体信号传导途径，和B）发展 BAD（Bcl-2相关死亡启动子）检测的改进方法， 细胞命运的调节因子，可被Akt磷酸化。 结果 获得的信息也可能有助于基本的更完整的 了解一个重要的临床实体，多囊卵巢综合征 （PCOS），其特征部分在于加速卵泡闭锁 （细胞凋亡）、胰岛素抵抗和男性化。