Post-transcriptional gene regulation in the ovary

描述(申请人提供)：促黄体生成素的排卵前激增触发排卵，颗粒细胞的黄体化和卵母细胞减数分裂的恢复，从而协调多种细胞和细胞外事件，形成“成熟”的卵母细胞和黄体。促黄体生成素引起的许多下游事件(即细胞信号、转录)已为人所知，但对排卵期卵泡中基因表达的转录后调控(mRNA剪接、降解、定位、翻译启动)知之甚少。这项研究的长期目标是确定在排卵周卵泡中颗粒细胞快速分化过程中转录后调节的作用，以确定转录和转录后过程如何介导排卵和黄体化。这项提议将“建立一种方法”来识别在体内经历独特转录后调控的基因转录本和生化途径，并将研究特定的核糖核蛋白(RNP)在排卵中所起的作用。AIM 1将通过对多聚体RNA的表达分析来分析与细胞翻译机制相关的基因(即间接的蛋白质合成)。目的2将使用特定RNPs的抗体，展示与一组相似的RNPs(S)相互作用的基因转录本的分离和鉴定。由于RNP协调转录后的过程并为mRNAs在细胞内提供地址(定位)，因此通过与特定RNP的关联将独特的基因转录本与特定的RNP相关联，应该允许我们在新的空间和时间背景下制定关于转录后调控的基本方面的假设。这项拟议的研究具有创新性，使用翻译(多聚体RNA)图谱方法来填补关键内分泌激素黄体生成素下游指导排卵和黄体化过程的基因的知识空白。这些使用小鼠模型的活体实验将首次描述哺乳动物系统中荷尔蒙调节的转录后机制。避孕和不孕不育是育龄妇女面临的两大生活质量选择。从成熟卵泡中排出可受精的卵母细胞是自然或辅助生殖策略的基础，也是许多不孕不育妇女被破坏的关键过程。抑制排卵也是避孕技术的主要目标。这些研究将极大地增强我们对促黄体激素如何调节排卵过程的了解。作为了解卵巢功能的基本贡献，这项研究的长期益处可能最终影响到为不孕不育妇女开发更有效的避孕方法和治疗策略。

DESCRIPTION (provided by applicant): The preovulatory surge of LH triggers ovulation, luteinization of granulosa cells and resumption of oocyte meiosis, thereby coordinating multiple cellular and extracellular events with the formation of a "mature" oocyte and corpus luteum. Many downstream events (i.e., cell signaling, transcription) elicited by LH are known, however, little is known about the post-transcriptional regulation of gene expression (mRNA splicing, degradation, localization, initiation of translation) in the periovulatory follicle. The long-term objective of this research is to define the role of post-transcriptional regulation that occurs during rapid differentiation of granulosa cells in the periovulatory follicle to ascertain how transcriptional and post-transcriptional processes mediate ovulation and luteinization. This proposal will "establish an approach" to identify gene transcripts and biochemical pathways undergoing unique post-transcriptional regulation in vivo and will investigate the role specific ribonucleoproteins (RNP) play in ovulation. Aim 1 will profile genes that are associated with the translation machinery of the cell (i.e., an indirect profile of protein synthesis) by expression analysis of polysomal RNA. Aim 2 will demonstrate isolation and identification of gene transcripts that interact with a similar set(s) of RNPs, using antibodies to specific RNPs. Because RNPs coordinate the processes following transcription and provide an address (localization) for mRNAs within the cell, the linkage of unique gene transcripts by association to a specific RNP should allow us to develop informed hypotheses regarding fundamental aspects of post-transcriptional regulation within a novel spatial and temporal context. The proposed research is innovative, in using a translation (polysomal RNA) profiling approach to fill the gaps in the knowledge of genes that direct the process of ovulation and luteinization downstream of the critical endocrine hormone, LH. These in vivo experiments using a mouse model will provide the first description of a hormonal-regulated post-transcriptional mechanism in a mammalian system to date. Contraception and infertility are two major quality of life choices facing women of a reproductive age. Ovulation of a fertilizable oocyte from a mature follicle is fundamental to natural or assisted reproductive strategies and is the critical process disrupted in many infertile women. Inhibition of ovulation is also a primary target for contraceptive technology. These studies will measurably enhance our knowledge of how luteinizing hormone regulates the ovulatory process. The long-term benefits of this research as a basic contribution to understanding ovarian function may ultimately impact the development of more effective contraceptive methods and treatment strategies for infertile women.