Neurotensin: A Novel Mediator of Ovulation

神经降压素：一种新型的排卵调节剂

• 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
• 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

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.

女人的生育能力取决于导致排卵的一系列级联。这种排卵中的缺陷 过程导致所有女性不育病中有25％，支持排卵在生殖中的重要作用 成功并提供了有力的前提来探索排卵过程的介体。我们提出了这一点 神经素是排卵过程的这些关键调节剂之一。 排卵前LH激增促进排卵，从而增加卵巢内介质 这种行为是为了释放卵母细胞。我们的新颖而令人兴奋的初步数据表明神经素（NTS）， 小神经肽是排卵的关键内部介体。这是基于我们的发现，即NTS是 人排卵叶的颗粒细胞中最差异表达的基因之一， 排卵性促性腺激素激增后15,000倍。同样，LH也迅速而显着增加了NT 来自啮齿动物和猴子排卵卵泡的颗粒细胞中的mRNA。重要的是，注射神经蛋白 进入猴子前卵泡中的抗体阻塞卵母细胞释放，导致黄体未破裂的卵泡， 与排卵失败一致。同样，缺乏NTS表达的小鼠形成了黄体化卵泡 在LH/HCG给药后，NTS受体拮抗剂抑制了新型体外卵母细胞的释放 小鼠排卵模型。但是，除了我们的初步数据外，有关临时性的尚无 NTS受体的表达或NTS在排卵和女性生育中的功能。这些问题将是 通过采用特殊背景的协作方法在本提案中解决，技术 专业知识和项目负责人的模型。我们将利用一个高影响的，翻译的模型 将在指定之前和三个 在HCG（早期，晚期和分解后）后完成体内和体外研究后进行小鼠进行的时代 和猴子。排卵叶中NTS和NTS受体表达的调节将被照亮 并在AIM 1中进行比较，猴子和小鼠在AIM 1中进行比较。 将植物的颗粒，theca，积云和内皮细胞在AIM 2中阐明。NTS的作用 在排卵以及其他关键的生殖过程中，将在小鼠和猴子中揭示使用小鼠 在AIM 3中缺乏NTS表达，NTS受体拮抗剂和NTS中和抗体。 该建议的是库里博士和达菲博士之间的协作互动，以解决NTS的功能 及其在跨多种物种（包括特殊人类卵巢组织）的期次培养物中的受体 和非人类灵长类动物的体内研究，这是一种与人类生殖高相关的动物模型 功能。这在理解卵母细胞的基本宗旨方面具有非凡的重要性 发布提供了一个基本平台，可以改善不育女性的预后或相反 提供靶标，以防止卵母细胞释放女性违规。