Molecular Pathways Controlling Ovarian Gene Expression

控制卵巢基因表达的分子途径

• 批准号: 7913605
• 负责人: CARLOS OSCAR STOCCO
• 金额: $ 22.53万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-02-01 至 2014-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7913605
• 关键词: Acetylation; Acute; Address; Antibodies; Antral; Apoptosis; Aromatase; Atretic Follicles; Biological; Biological Assay; Cell Culture Techniques; Cell Death; Cell Proliferation; Cell physiology; Cells; Code; Collaborations; Defect; Development; Differentiation and Growth; Dominant-Negative Mutation; Electrophoretic Mobility Shift Assay; Embryo; Enzymes; Estradiol; Etiology; Female; Fertility; Follicle Stimulating Hormone; Follicle Stimulating Hormone Receptor; GATA4 transcription factor; Gene Expression; Gene Targeting; Genes; Glycogen Synthase Kinase 3; Gonadal Steroid Hormones; Growth; Growth Factor; Hormones; Immunohistochemistry; Immunoprecipitation; In Vitro; Infertility; Investigation; Knockout Mice; Knowledge; Light; Lysine; MAPK14 gene; Mediating; Modification; Molecular; Mus; Ovarian; Ovarian Cycles; Ovarian Diseases; Ovarian Granulosa Cell; Ovary; Paper; Pathway interactions; Peptides; Phosphorylation; Phosphotransferases; Physiological; Pituitary Hormones; Play; Production; Proliferating; Protein C; Proteins; Publishing; Regulation; Regulator Genes; Role; Small Interfering RNA; Staging; Steroid biosynthesis; Steroids; System; Techniques; Technology; Transcription Coactivator; Transgenic Model; Western Blotting; Woman; base; cofactor; design; folliculogenesis; gene repression; granulosa cell; granulosa cell tumor; improved functioning; in utero; in vivo; inhibin; intraovarian; mutant; oocyte maturation; overexpression; public health relevance; response; transcription factor

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. PUBLIC HEALTH RELEVANCE: The roles of the transcription factors GATA-4 and GATA-6 in folliculogenesis and fertility will be addressed. These studies will contribute to better understanding of the cellular and molecular mechanisms that control ovarian function and improve our knowledge of the etiology of ovarian diseases 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，这些转录因子被证明在涉及类固醇生成的酶的表达中起关键作用。我们在最近发表的几篇论文中证明了GATA-4对于雌二醇的产生至关重要，这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有关的发现表明，该因素在卵巢中也起生存作用。由于GATA-4和GATA-6无效小鼠是胚胎致死的，因此GATA在卵巢功能中的最终作用仍然未知。因此，这项研究旨在确定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。这项研究的具体目的是：1-生成卵巢特异性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的编码区域的一部分或它们的组合。我们预计这些小鼠在类固醇产生和卵泡发育方面会有严重的缺陷。 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特定作用。 3-鉴定在体内功能性卵巢中受GATA-4和GATA-6调节的基因，我们将使用芯片片上的芯片分析。迄今为止获得的所有结果表明，GATA-4和GATA-6在卵巢的正常功能中可能起着至关重要的作用。我们期望这项研究清楚地确定GATA-4和GATA-6在颗粒细胞的类固醇生成能力和生长中的功能，以确定GATA-4和GATA-6是否对这些细胞具有冗余作用，并鉴定GATA-4和GATA-4和GATA-6靶基因。这项研究不仅将提供有关正常卵巢功能的调节机制的至关重要的信息，而且由于颗粒细胞功能不当，例如颗粒细胞肿瘤和不孕症，可能会阐明卵巢疾病。公共卫生相关性：将解决转录因子GATA-4和GATA-6在卵泡发生和生育方面的作用。这些研究将有助于更好地理解控制卵巢功能的细胞和分子机制，并提高我们对卵巢疾病（例如颗粒细胞肿瘤和不育）病因的了解。