Molecular Pathways Controlling Ovarian Gene Expression

控制卵巢基因表达的分子途径

基本信息

批准号：
8431437
负责人：
CARLOS OSCAR STOCCO
金额：
$ 27.4万
依托单位：
UNIVERSITY OF ILLINOIS AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-02-01 至 2015-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8431437
关键词：
Achievement Activins Address Adenoviruses Alleles Androgens Animals Apoptosis Apoptotic Aromatase Biological Assay CYP17A1 gene Cell Culture Techniques Cell Cycle Progression Cell Death Cell Proliferation Cell Survival Cell physiology Cells ChIP-on-chip Code DNA Defect Development Embryo Enzymes Estradiol Estrogens Etiology Female Fertility Gene Expression Gene Expression Regulation Gene Targeting Genes Genetic Recombination Genomics Granulosa-Lutein Cells Growth Health Hormones In Vitro Infection Infertility Investigation Knockout Mice Knowledge Light Microarray Analysis Molecular Mus Ovarian Ovarian Cycles Ovarian Diseases Ovarian Granulosa Cell Ovary Paper Pathway interactions Physiological Play Production Publishing Reproduction Reproductive Process Role Staging Steroid biosynthesis Steroids Structure Technology Testing design folliculogenesis gene repression granulosa cell granulosa cell tumor improved functioning in vivo mutant overexpression promoter recombinase response theca cell transcription factor

项目摘要

DESCRIPTION (provided by applicant): Coordinated activation and repression of genes are essential for the normal progress of the ovarian cycle. Ovarian granulosa cells express high levels of the transcription factors GATA-4 and GATA-6. In cell cultures, these transcription factors were shown to play key roles in the expression of enzymes involved in steroidogenesis. We have demonstrated in several recently published papers that GATA-4 is critical for the production of estradiol, which is essential for granulosa cell proliferation and follicle maturation. In addition, the finding that low expression of GATA-4 is associated with follicular cell death suggests that this factor also plays a survival role in the ovary. Because GATA-4 and GATA-6 null mice are embryonic lethal, the ultimate role of GATA in ovarian function remains unknown. This study is therefore designed to determine the physiological role of GATA-4 and GATA-6 in ovarian steroidogenesis, follicular development, and female fertility. The specific aims of this study are: 1- To generate ovarian-specific GATA-4 and GATA-6 null mice in order to find out the explicit roles of these transcription factors in the ovary in vivo. The Cre-lox technology will be used to delete part of the coding region of GATA-4, GATA-6, or their combination. We expect these mice to have severe defects in steroid production and in follicular development. 2- To determine the effect of GATA-4 and GATA-6 deletion on ovarian cells in vitro. Granulosa cells lacking GATA-4, GATA-6, or both will be used to examine the specific roles of these factors in steroidogenesis, cell proliferation, granulosa-cell hormone response, and gene expression. 3- To identify those genes that are regulated by GATA-4 and GATA-6 in fully functional ovaries in vivo, we will use ChIP-on-chip assays. All the results obtained to date indicate that GATA-4 and GATA-6 may play a crucial role in the normal function of the ovary. We expect from this investigation to clearly define the function of GATA-4 and GATA-6 in the steroidogenic capacity and growth of granulosa cells, to determine whether GATA-4 and GATA-6 have redundant effects on these cells, and to identify GATA-4 and GATA-6 target genes in the ovary. This study will not only provide critically important information on the regulatory mechanism underlying normal ovarian function but also may shed light on ovarian diseases due to improper granulosa cell function such as granulosa cell tumors and infertility.

描述（由申请人提供）：基因的协调激活和抑制对于卵巢周期的正常进展至关重要。卵巢颗粒细胞表达高水平的转录因子 GATA-4 和 GATA-6。在细胞培养物中，这些转录因子在类固醇生成相关酶的表达中发挥着关键作用。我们在最近发表的几篇论文中证明，GATA-4 对于雌二醇的产生至关重要，而雌二醇对于颗粒细胞增殖和卵泡成熟至关重要。此外，GATA-4低表达与卵泡细胞死亡相关的发现表明该因子也在卵巢中发挥着生存作用。由于 GATA-4 和 GATA-6 缺失小鼠是胚胎致死的，因此 GATA 在卵巢功能中的最终作用仍不清楚。因此，本研究旨在确定 GATA-4 和 GATA-6 在卵巢类固醇生成、卵泡发育和女性生育能力中的生理作用。本研究的具体目的是： 1- 产生卵巢特异性GATA-4和GATA-6无效小鼠，以找出这些转录因子在体内卵巢中的明确作用。 Cre-lox技术将用于删除GATA-4、GATA-6或其组合的部分编码区。我们预计这些小鼠在类固醇产生和卵泡发育方面存在严重缺陷。 2-确定GATA-4和GATA-6缺失对体外卵巢细胞的影响。缺乏 GATA-4、GATA-6 或两者的颗粒细胞将用于检查这些因子在类固醇生成、细胞增殖、颗粒细胞激素反应和基因表达中的具体作用。 3- 为了识别体内功能齐全的卵巢中受 GATA-4 和 GATA-6 调节的基因，我们将使用 ChIP 芯片检测。迄今为止获得的所有结果表明GATA-4和GATA-6可能在卵巢的正常功能中发挥着至关重要的作用。我们希望通过这项研究清楚地定义 GATA-4 和 GATA-6 在类固醇生成能力和颗粒细胞生长中的功能，确定 GATA-4 和 GATA-6 是否对这些细胞有多余的影响，并确定 GATA- 4 和 GATA-6 卵巢中的靶基因。这项研究不仅将提供关于正常卵巢功能的调节机制的极其重要的信息，而且还可能揭示由于颗粒细胞功能不当引起的卵巢疾病，例如颗粒细胞瘤和不孕症。