Genetic interrogation of conserved follicular factors for matrix metalloproteinase regulation and ovulation

基质金属蛋白酶调节和排卵的保守卵泡因子的遗传询问

• 批准号: 9269226
• 负责人: Jianjun Sun
• 金额: $ 33.51万
• 依托单位: UNIVERSITY OF CONNECTICUT STORRS
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-05-04 至 2021-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9269226
• 关键词: Anovulation; Biological Assay; Biological Models; Calcium; Cells; Complex; Contraceptive Agents; Contraceptive methods; Data; Development; Disease; Drosophila genus; Encapsulated; Event; Future; Genes; Genetic; Genetic Screening; Graafian Follicles; Health; Human; Infertility; Investigation; Irregular Menstruation; Knowledge; Luteinization; MAPK8 gene; Mammalian Oviducts; Mammals; Matrix Metalloproteinases; Methods; Modeling; Molecular; Mus; N-terminal; NADPH Oxidase; Nature; Nuclear Receptors; Oocytes; Orthologous Gene; Ovarian; Ovarian Follicle; Ovulation; Oxidative Stress; Pathway interactions; Pharmacology; Phosphotransferases; Play; Process; Production; Reactive Oxygen Species; Regulation; Role; Rupture; Signal Pathway; Signal Transduction; Signal Transduction Pathway; System; Testing; Woman; Work; calmodulin-dependent protein kinase II; fly; gene function; genetic approach; genetic manipulation; granulosa cell; in vivo; infertility treatment; insight; new therapeutic target; novel; oocyte maturation; programs; protein expression; public health relevance; spatiotemporal; tool

 DESCRIPTION (provided by applicant): Follicle rupture is the final step of the complex ovulation program, which releases fertilizable oocytes. Despite intensive study in the past four decades, a comprehensive understanding of the molecular mechanisms of follicle rupture is still lacking, in part due to the limitation of mammalian model systems to utilize genetic screens. We recently developed a novel Drosophila system that allows rapid application of genetic approaches to reveal precise details regarding the molecular events of follicular rupture. Moreover, recent studies from our lab have shown that the basic cellular and molecular mechanisms of ovulation are highly conserved from flies to humans; for instance, both systems require matrix metalloproteinase (Mmp) activity for follicle rupture. Leveraging the wealth of genetic tools and our ex vivo ovulation assay, this project will systematically interrogate conserved factors that are required for the precise regulation of Mmp activity and follicle rupture Our preliminary data reveals that an increase in intracellular free Ca2+ is required for Mmp activation but not expression. Conversely, follicular NADPH oxidase (Nox), which generates reactive oxygen species (ROS), and the oxidative stress-induced c-Jun N-terminal kinase (JNK) pathway regulate spatiotemporal Mmp protein expression. Therefore, we propose to 1) elucidate the conserved calcium-dependent signal transduction pathways that are required for Mmp activation, 2) investigate the role of follicular ROS and JNK signaling in Mmp expression, and 3) identify novel follicular factors for Mmp regulation and ovulation using genetic screens. This work will provide a comprehensive understanding of the ovarian signaling networks that precisely regulate Mmp activity and follicle rupture, which would be difficult in mammalian model systems. The conserved nature of these signaling pathways will allow the knowledge gained from this study to be further validated in mammalian and human ovulation. Therefore, this work will ultimately reveal promising new drug targets for the alleviation of anovulatory infertility orfor contraceptive development, both of which are highly relevant to human health.

 描述（通过应用程序提供）：卵泡破裂是复杂排卵程序的最后一步，该程序释放了可肥大的卵母细胞。尽管在过去的四十年中进行了深入的研究，但仍然缺乏对叶叶片破裂的分子机制的全面了解，部分原因是哺乳动物模型系统利用遗传筛查的限制。我们最近开发了一种新型的果蝇系统，该系统允许快速应用遗传方法，以揭示有关卵泡破裂分子事件的精确细节。此外，我们实验室的最新研究表明，排卵的基本细胞和分子机制是从蝇到人的高度保守的。例如，这两个系统都需要基质金属蛋白酶（MMP）活性才能进行Follic破裂。利用遗传工具和我们的离体排卵测定法，该项目将系统地询问保守的因素，这些因素是对MMP活性和Follice破裂的精确调节所必需的，我们的初步数据表明，MMP激活所需的细胞内游离CA2+的增加，但不表达。相反，产生活性氧（ROS）以及氧化应激诱导的C-JUN N末端激酶（JNK）途径的卵泡氧化物（NOX）调节时空MMP蛋白表达。因此，我们建议1）阐明MMP激活所需的钙依赖性信号转导途径，2）研究卵泡ROS和JNK信号在MMP表达中的作用，3）识别使用遗传筛查的MMP调控和排卵的新型卵泡因子。这项工作将对卵巢信号网络进行全面的了解，该网络准确地调节MMP活性和卵泡破裂，这在哺乳动物模型系统中很难。这些信号通路的配置性质将允许在哺乳动物和人类排卵中进一步验证这项研究的知识。因此，这项工作最终将揭示有望减轻发育不育症或避孕发展的新药物目标，这两者都与人类健康高度相关。