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LH Action in Ovarian Cell Differentiation

LH 在卵巢细胞分化中的作用

基本信息

批准号：
8495779
负责人：
JoAnne Stewart Richards
金额：
$ 34.61万
依托单位：
BAYLOR COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
1982
资助国家：
美国
起止时间：
1982-03-01 至 2014-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8495779
关键词：
AREG gene Aromatase Biological Process Cattle Cell Differentiation process Cell physiology Cells Complement 1q Complex Data Development EGF gene EREG gene Embryo Endocrine Epidermal Growth Factor Receptor Event Exhibits Extracellular Matrix Fertility Fertilization Follicle Stimulating Hormone Gene Expression Gene Targeting Generations Genes Genetic Programming Gonadotropins Health Human Hyaluronan Immune Infertility Inflammation Inflammatory Inflammatory Response Interleukin-6 Knockout Mice Ligands Luteinizing Hormone MAPK1 gene MAPK3 gene Mammalian Oviducts Mediating Mediator of activation protein Menstrual cycle Molecular Molecular Profiling Mus Mutant Strains Mice Oocytes Ovarian Ovarian Follicle Ovary Ovulation Pathway interactions Process Production Prostaglandins Reaction Role Signal Transduction Signaling Molecule Surface System TLR2 gene TLR4 gene Testing Toll-like receptors Woman base cytokine design granulosa cell in vivo in vivo Model mouse model mutant mouse model novel oocyte maturation promoter receptor response

项目摘要

DESCRIPTION (provided by applicant): Ovulation is the unique biological process by which a mature oocyte and surrounding cumulus cells, comprising the cumulus cell-oocyte complex (COC), are released from the surface of the ovary. This LH-induced process is similar to an inflammatory response and requires the expression of specific genes: the EGF-like factors (Areg, Ereg, Btc), matrix factors (Has2, Ptgs2, Tnfaip6, Ptx3) and cytokines (Il6). AREG potently activates ERK1/2, COC expansion, oocyte maturation and IL6 production in cultured COCs. However, the genes regulated specifically by AREG/ERK1/2 and the consequences of disrupting these signaling molecules in vivo have not been determined. Therefore, using Erk1 (Erk1-/-) null mice, Erk2fl/fl mice and mice expressing Cre driven by the aromatase (Cyp19) promoter, we have generated Erk1/2 double KO (Erk1-/- ;Erk2fl/fl/;Cyp19-Cre) mice. Follicular development is normal but preovulatory follicles fail to ovulate and COC expansion is impaired in the Erk1/2 double KO mice. In addition to inflammation-related genes, LH/AREG also regulate the expression of specific genes that comprise the innate-immune cell surveillance response system, including the Toll-like receptors TLR2 and TLR4 and components of this system (C1q, Cd14, and Pdcd1) uniquely expressed in cumulus cells. TLR2 and TLR4 are functional in cumulus cells and respond to known ligands (including matrix factors) leading to the production of IL6 that can induce COC expansion in culture in the absence of AREG or PGE. Preliminary data indicate that Tlr4-/- mice are subfertile. By analyzing the signaling cascades and the expression of specific genes in Erk1/2 double KO mice and in mice null for the Areg target genes (Tlr/42, Ptx3 and Pdcd1) that are expressed in COCs, we should be able to dissect and delineate a novel genetic program that controls cumulus cell function and ovulation. Because the production of cytokines, including IL6, appears to be critical for ovulation, we propose that the ERK1/2 pathway controls an endocrine-immune-cytokine regulatory network during ovulation. Therefore we propose these specific aims: Specific Aim I: Determine the molecular mechanisms by of ERK2 (MAPK1) and ERK1 (MAPK3) regulate granulosa cell and cumulus cell functions and ovulation in vivo. Test the hypothesis that ERK1/2 mediate specific effects of LH induced ovulation and cumulus cell function. Specific Aim II: Determine the function of AREG (ERK1/2) target genes that are uniquely expressed at high levels in ovulated COCs and that impact ovulation. Test the hypotheses that specific matrix factors, IL6 and innate immune cell-related genes expressed in cumulus cells direct the unique fate of these cells.

描述(申请人提供)：排卵是一种独特的生物过程，成熟的卵母细胞和周围的卵丘细胞，组成卵丘细胞-卵母细胞复合体(COC)，从卵巢表面释放出来。促黄体生成素诱导的这一过程类似于炎症反应，需要特定基因的表达：EGF样因子(Areg、Ereg、BTC)、基质因子(Has2、Ptgs2、Tnfaip6、Ptx3)和细胞因子(IL6)。AREG能有效地激活培养的COCs的ERK1/2、COC扩增、卵母细胞成熟和IL6的产生。然而，AREG/ERK1/2特异调控的基因以及体内干扰这些信号分子的后果尚未确定。因此，利用ERK1(ERK1-/-)缺失小鼠、Erk2fl/fl小鼠和芳香酶(Cyp19)启动子驱动表达Cre的小鼠，我们获得了ERK1/2双KO(ERK1-/-；Erk2fl/fl/；Cyp19-Cre)小鼠。ERK1/2双KO小鼠卵泡发育正常，但排卵前卵泡无法排卵，COC扩张受损。除了炎症相关基因，黄体生成素/AREG还调节组成天然免疫细胞监视反应系统的特定基因的表达，包括Toll样受体TLR2和TLR4以及该系统的组件(C1q、CD14和Pdcd1)在卵丘细胞中唯一表达。TLR2和TLR4在卵丘细胞中发挥作用，并对已知的配体(包括基质因子)做出反应，导致IL6的产生，在没有AREG或PGE的情况下，IL6可以在培养中诱导COC扩张。初步数据表明，TLR4-/-小鼠是不育的。通过分析ERK1/2双KO小鼠和在COC中表达的Areg靶基因(TLR/42、Ptx3和Pdcd1)缺失的小鼠的信号级联和特定基因的表达，我们应该能够解剖和描绘控制卵丘细胞功能和排卵的新的遗传程序。由于包括IL6在内的细胞因子的产生似乎对排卵至关重要，我们认为ERK1/2途径在排卵过程中控制着内分泌-免疫-细胞因子调节网络。因此，我们提出了这些特定的目的：特定目的I：确定ERK2(MAPK1)和ERK1(MAPK3)在体内调节颗粒细胞和卵丘细胞功能和排卵的分子机制。验证ERK1/2介导促黄体生成素诱导排卵和卵丘细胞功能的特异性效应的假设。特定目的II：确定AREG(ERK1/2)靶基因的功能，这些基因在排卵的COCs中唯一高水平表达，并影响排卵。测试在卵丘细胞中表达的特定基质因子、白介素6和先天免疫细胞相关基因决定这些细胞独特命运的假设。