Neurotensin: A Novel Mediator of Ovulation

神经降压素：一种新型的排卵调节剂

• 批准号: 10657389
• 负责人: Thomas E Curry
• 金额: $ 64.5万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-06 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10657389
• 关键词: Address; Agonist; Animal Model; Antibodies; Bioinformatics; Cell physiology; Central Nervous System; Complement; Data; Defect; Development; EGF gene; Endothelial Cells; Evaluation; Event; Failure; Fc Receptor; Female Contraceptive Agents; Female infertility; Fertility; Fertilization; Future; Gene Expression Profiling; Genes; Gonadotropins; Histologic; Human; Human Chorionic Gonadotropin; In Vitro; Infertility; Injections; LH Receptors; Luteinization; Mediating; Mediator; Messenger RNA; Modeling; Monkeys; Motion; Mus; Neuropeptides; Neurotensin; Neurotensin Receptors; New Agents; Oocytes; Ovarian; Ovarian Follicle; Ovarian Tissue; Ovary; Ovulation; Pathway interactions; Play; Pregnancy; Pregnancy Outcome; Process; Progesterone; Prostaglandins; Receptor Signaling; Regulation; Reproductive Process; Reproductive system; Rodent; Role; Rupture; Signal Transduction; System; Technical Expertise; Testing; Time; Vascular Endothelial Cell; Wild Type Mouse; Woman; angiogenesis; antagonist; cell type; collaborative approach; differential expression; experimental study; female fertility; granulosa cell; implantation; improved outcome; in vitro Model; in vivo; mouse model; neutralizing antibody; nonhuman primate; novel; oocyte quality; paracrine; prevent; receptor; receptor expression; reproductive function; reproductive success; response; theca cell; translational model

A woman’s fertility is dependent on the cascade of events leading up to ovulation. Defects in this ovulatory process result in 25% of all female infertility cases, supporting the paramount role of ovulation in reproductive success and providing a strong premise to explore mediators of the ovulatory process. We propose that neurotensin is one of these pivotal regulators of the ovulatory process. Ovulation is set in motion by the preovulatory LH surge which increases ovarian intrafollicular mediators that act to bring about oocyte release. Our novel and exciting preliminary data indicate that neurotensin (NTS), a small neuropeptide, is a key intrafollicular mediator of ovulation. This is based upon our findings that NTS is one of the most differentially-expressed genes in the granulosa cells of the human ovulatory follicle, up to 15,000 fold after the ovulatory gonadotropin surge. Likewise, LH also rapidly and significantly increases NTS mRNA in granulosa cells from rodent and monkey ovulatory follicles. Importantly, injection of a neurotensin antibody into monkey preovulatory follicles blocked oocyte release resulting in luteinized unruptured follicles, consistent with failure of ovulation. Similarly, mice lacking NTS expression form luteinized unruptured follicles following LH/hCG administration, and a NTS receptor antagonist inhibits oocyte release in a novel in vitro model of mouse ovulation. Yet, other than our preliminary data, nothing is known regarding the temporal expression of NTS receptors or the function of NTS in ovulation and female fertility. These questions will be addressed in this proposal through a collaborative approach utilizing the exceptional backgrounds, technical expertise, and models of the Project Leaders. We will make use of a high-impact, translational model where the granulosa and theca from human periovulatory follicles will be collected prior to and at three designated times after hCG (early, late and postovulatory) to complement in vivo and in vitro studies performed with mice and monkeys. The regulation of NTS and NTS receptor expression in the ovulatory follicle will be illuminated and compared across women, monkeys and mice in Aim 1. The ovulatory functions stimulated by NTS in granulosa, theca, cumulus, and endothelial cells of the follicle will be elucidated in Aim 2. The actions of NTS on ovulation, as well as other key reproductive processes, will be revealed in mice and monkeys using mice lacking NTS expression, NTS receptor antagonists, and NTS-neutralizing antibodies in Aim 3. A major strength of this proposal is the collaborative interactions between Dr. Curry and Dr. Duffy to address the function of NTS and its receptors in periovulatory events across multiple species, including exceptional human ovarian tissues and in vivo studies of non-human primates, an animal model with high relevance to human reproductive function. Thus, there is an exceptional degree of significance in understanding the basic tenets of oocyte release which provides a fundamental platform to either improve outcomes for infertile women or conversely provide targets to prevent oocyte release for female contraception.

女性的生育能力取决于导致排卵的一系列事件。这个排卵器有缺陷 过程导致25%的女性不孕，支持排卵在生殖中的重要作用 并为探索排卵过程的介体提供了有力的前提。我们建议 神经降压素是排卵过程中的关键调节因子之一。 排卵是由排卵前的促黄体生成素激增启动的，它增加了卵泡内介质。 导致卵子释放的行为。我们新颖而令人兴奋的初步数据表明，神经降压素(NTS)， 一种小的神经肽，是卵泡内排卵的关键介质。这是基于我们的发现，NTS是 在人类排卵卵泡的颗粒细胞中差异表达最多的基因之一， 排卵期促性腺激素激增后15,000倍。同样，促黄体生成素也迅速而显著地增加了NTS 啮齿动物和猕猴排卵卵泡颗粒细胞中的mRNA。重要的是，注射神经降压素 进入猴子排卵前卵泡的抗体阻止了卵母细胞的释放，导致黄素化的未破裂卵泡， 与排卵失败相一致。同样，缺乏NTS表达的小鼠形成黄素化的未破裂卵泡 在给予黄体生成素/人绒毛膜促性腺激素后，NTS受体拮抗剂在体外抑制卵母细胞的释放 小鼠排卵模型。然而，除了我们的初步数据，关于时间上的 NTS受体的表达及其在排卵和女性生育中的作用这些问题将是 在本建议书中通过利用特殊背景、技术 项目负责人的专业知识和模式。我们将使用高影响力的翻译模式，其中 人类排卵周围卵泡的颗粒和卵膜将在指定的三个月之前和之后收集 在小鼠体内和体外进行的hCG补体时间(早期、晚期和排泄后)研究 还有猴子。将阐明NTS及其受体在排卵卵泡中的表达调控。 并在目标1中对女性、猴子和小鼠进行了比较。NTS刺激的排卵功能 目标2将阐明卵泡的颗粒、膜、卵丘和内皮细胞。NTS的作用 关于排卵以及其他关键生殖过程的研究将在小鼠和猴子身上揭示。 在AIM中缺乏NTS表达、NTS受体拮抗剂和NTS中和抗体3.主要优势 这项提议的一部分是库里博士和达菲博士之间的合作互动，以解决NTS的功能 及其受体在多个物种的排卵事件中的作用，包括特殊的人类卵巢组织 对非人类灵长类动物的活体研究，这是一种与人类生殖高度相关的动物模型 功能。因此，了解卵母细胞的基本原理具有非常重要的意义。 释放，为改善不孕妇女的结局或相反提供了一个基本的平台 为女性避孕提供防止卵母细胞释放的目标。