Leukemia Inhibitory Factor As a Mediator of Primate Ovulation & Oocyte Maturation

白血病抑制因子作为灵长类动物排卵的调节剂

基本信息

批准号：
8554777
负责人：
Jon D Hennebold
金额：
$ 20.76万
依托单位：
OREGON HEALTH & SCIENCE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-28 至 2015-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8554777
关键词：
Animals Area Assisted Reproductive Technology Binding Binding Proteins Bolus Infusion Cell physiology Cell surface Cells Complex Contraceptive methods Data Databases Development Embryo Embryonic Development Estradiol Evaluation Event Extracellular Matrix Family member Fertilization Fertilization in Vitro Follicle Stimulating Hormone Follicular Fluid Gene Expression Genes Genomics Glycoproteins Gonadotropins Guidelines Histologic Human Human Chorionic Gonadotropin Hyaluronan In Vitro Individual Infertility Injection of therapeutic agent JAK1 gene Janus kinase Janus kinase 1 Knowledge LIFR gene Lead Length Luteal Phase Luteinizing Hormone Macaca Macaca mulatta Measures Mediator of activation protein Meiosis Menstruation Messenger RNA Modeling Molecular Monitor Multiple Birth Offspring Mus Nuclear Oocytes Ovarian Follicle Ovarian Stimulations Ovary Ovulation Ovulation Induction Ovum PTGS2 gene Pathway interactions Phosphorylation Pilot Projects Pituitary Gland Play Primates Process Progesterone Protein Tyrosine Kinase Proteins Protocols documentation Publishing Recombinants Risk Role Rupture STAT3 gene Signal Transduction Stat3 protein Stimulus TYK2 Techniques Testing Time Treatment Protocols Tumor Necrosis Factor-alpha corpus luteum cyclooxygenase 2 cytokine extracellular granulosa cell implantation improved in vivo leukemia inhibitory factor leukemia inhibitory factor receptor novel oocyte maturation paracrine prevent receptor reproductive success research study social stigma transcription factor

项目摘要

DESCRIPTION (provided by applicant): The cellular and molecular processes occurring within the primate follicle resulting in the release of a mature ovum (ovulation) that fertilizes and undergoes subsequent embryonic development are incompletely defined. Systematic and detailed characterizations of such events are necessary for advancing infertility treatments and developing novel, non-hormonal forms of contraception. In this regard, studies conducted by the P.I. using a high-throughput genomic approach led to the identification of most, if not all, genes whose expression increases through the periovulatory interval following an ovulatory stimulus. Such mRNAs are likely involved in activities necessary for follicle rupture, which include the formation of a hyaluronan-rich extracellular matrix between cumulus cells and the loss of their cell-cell contacts (cumulus-oocyte expansion; C-OE), as well as the cytoplasmic and nuclear maturation of the oocyte required for subsequent fertilization and embryonic development. From the resultant database and additional preliminary studies, it was discovered that leukemia inhibitory factor (LIF) mRNA and protein increased in the rhesus macaque follicle from undetectable levels before administration of an ovulatory stimulus (human chorionic gonadotropin; hCG; 0 h controls) to peak values prior to (12 h post-hCG) and following ovulation (36 h post-hCG). Furthermore, mRNAs encoding downstream signaling components responsible for LIF action (glycoprotein 130, or gp130; janus kinase 1, or JAK1; signal transducer and activator of transcription 3, or STAT3) were also highest in unruptured rhesus macaque follicles 12 h after hCG administration and those that had ovulated 36 h post-hCG. The mRNAs encoding both cell surface LIF binding proteins (LIF receptor, or LIFR; gp130) were further localized to isolated oocytes and granulosa cells. Lastly, in pilot studies, intrafollicula injection of a LIF antagonist (the extracellular LIF binding portion of the LIFR; referred to as soluble LIFR or sLIFR) prevents rupture of the rhesus macaque follicle following an ovulatory stimulus, whereas injection of vehicle alone results in ovulation. Collectively, these findings support the hypothesis that LIF synthesis and signaling in the primate ovary is a critical regulator of events necessary for ovulation and formation of the corpus luteum (assessed in Aim 1); as well as for C-OE, reinitiation of oocyte meiosis, fertilization, and early embryonic development (assessed in Aim 2). Novel techniques involving intrafollicular injection of a LIF antagonist (sLIFR) will be employed to assess the role LIF plays in follicle rupture as well the formation of the corpus luteum (i.e., the ability to synthesize progesterone and estradiol). Isolated rhesus macaque cumulus-oocyte complexes (COCs) from non-luteinized follicles (i.e., pre-hCG) will be directly exposed to LIF to determine a direct effect of this cytokine on promoting C-OE, reinitiation of meiosis, fertilization, and embryonic development.

描述（由申请人提供）：灵长类动物卵泡内发生的导致释放成熟卵子（排卵）、受精并经历随后的胚胎发育的细胞和分子过程尚未完全定义。对此类事件进行系统和详细的描述对于推进不孕症治疗和开发新型非激素避孕方式是必要的。在这方面，P.I. 进行的研究使用高通量基因组方法识别出大多数（如果不是全部）基因，这些基因的表达在排卵刺激后的排卵期间隔期间增加。此类 mRNA 可能参与卵泡破裂所需的活动，其中包括卵丘细胞之间富含透明质酸的细胞外基质的形成以及细胞与细胞接触的丧失（卵丘-卵母细胞扩张；C-OE），以及随后受精和胚胎发育所需的卵母细胞的细胞质和核成熟。从所得数据库和其他初步研究中发现，恒河猴卵泡中的白血病抑制因子 (LIF) mRNA 和蛋白质从施用排卵刺激剂（人绒毛膜促性腺激素；hCG；0 小时对照）之前的不可检测水平增加到峰值排卵前（hCG 后 12 小时）和排卵后（hCG 后 36 小时）的值。此外，编码负责 LIF 作用的下游信号成分（糖蛋白 130 或 gp130；janus 激酶 1 或 JAK1；信号转导器和转录激活剂 3 或 STAT3）的 mRNA 在 hCG 给药后 12 小时在未破裂的恒河猴卵泡中也最高。那些在 hCG 后 36 小时排卵的人。编码细胞表面 LIF 结合蛋白（LIF 受体或 LIFR；gp130）的 mRNA 进一步定位于分离的卵母细胞和颗粒细胞。最后，在初步研究中，卵泡内注射 LIF 拮抗剂（LIFR 的细胞外 LIF 结合部分；称为可溶性 LIFR 或 sLIFR）可防止恒河猴卵泡在排卵刺激后破裂，而单独注射媒介物会导致排卵。总的来说，这些发现支持这样的假设：灵长类动物卵巢中的 LIF 合成和信号传导是排卵和黄体形成所需事件的关键调节因子（目标 1 中评估）；对于 C-OE，卵母细胞减数分裂、受精和早期胚胎发育的重新启动（在目标 2 中评估）。将采用涉及卵泡内注射 LIF 拮抗剂 (sLIFR) 的新技术来评估 LIF 在卵泡破裂以及黄体形成（即合成孕酮和雌二醇的能力）中的作用。来自非黄素化卵泡（即 hCG 前）的分离恒河猴卵丘卵母细胞复合物 (COC) 将直接暴露于 LIF，以确定该细胞因子对促进 C-OE、减数分裂重新启动、受精和胚胎的直接影响发展。