Neurotensin: A Novel Mediator of Ovulation

神经降压素：一种新型的排卵调节剂

基本信息

批准号：
10011839
负责人：
Thomas E Curry
金额：
$ 67.55万
依托单位：
UNIVERSITY OF KENTUCKY
依托单位国家：
美国
项目类别：
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/10011839
关键词：
Address Agonist Animal Model Antibodies Bioinformatics Cell physiology 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 of activation protein Messenger RNA Modeling Monkeys Motion Mus Neuraxis 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 base 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

项目摘要

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。该过程导致 25% 的女性不孕症病例，支持排卵在生殖中的重要作用成功并为探索排卵过程的中介提供了强有力的前提。神经降压素是排卵过程的关键调节剂之一。排卵前 LH 激增会增加卵巢卵泡内介质，从而启动排卵我们新颖且令人兴奋的初步数据表明神经降压素（NTS），一种小神经肽，是卵泡内排卵的关键介质。这是基于我们的发现，NTS 是。人类排卵卵泡颗粒细胞中表达差异最大的基因之一，高达排卵促性腺激素激增后 15,000 倍，类似地，LH 也会迅速而显着地增加 NTS。啮齿动物和猴子排卵卵泡颗粒细胞中的 mRNA 重要的是，注射神经降压素。猴抗体进入排卵前卵泡阻止卵母细胞释放，导致未破裂卵泡黄素化，与排卵失败一致。缺乏 NTS 表达的小鼠形成黄素化的类似未破裂的卵泡 LH/hCG 给药后，NTS 受体拮抗剂在新型体外抑制卵母细胞释放然而，除了我们的初步数据之外，我们对时间一无所知。 NTS 受体的表达或 NTS 在排卵和女性生育能力中的功能。本提案通过协作方式解决了这一问题，利用了特殊的背景、技术我们将利用具有高影响力的转化模型。来自人类排卵期卵泡的颗粒层和卵泡膜将在指定的三个时间点之前和时间收集 hCG 后的时间（排卵早期、晚期和排卵后）以补充小鼠体内和体外研究和猴子排卵卵泡中 NTS 和 NTS 受体表达的调节将得到阐明。并在目标 1 中对女性、猴子和小鼠进行比较。NTS 刺激的排卵功能目标 2 将阐明卵泡的颗粒细胞、卵泡膜、卵丘和内皮细胞。NTS 的作用排卵以及其他关键生殖过程的影响将在小鼠和猴子中使用小鼠来揭示目标 3 中缺乏 NTS 表达、NTS 受体拮抗剂和 NTS 中和抗体。主要优势该提案的核心是 Curry 博士和 Duffy 博士之间的协作互动，以解决 NTS 的功能及其受体在多个物种的排卵期事件中的作用，包括特殊的人类卵巢组织以及非人类灵长类动物的体内研究，这是一种与人类生殖高度相关的动物模型因此，了解卵母细胞的基本原理具有特殊的意义。发布，为改善不孕妇女的结果或相反提供了一个基本平台为女性避孕提供防止卵母细胞释放的目标。