Post-transcriptional gene regulation in the ovary

描述（由申请人提供）：排卵前LH的激增触发排卵，颗粒细胞的黄体化和卵母细胞减数分裂的恢复，从而协调多个细胞和细胞外事件与“成熟”卵母细胞和黄体的形成。LH引发的许多下游事件（即细胞信号传导、转录）是已知的，然而，对卵泡内基因表达的转录后调控（mRNA剪接、降解、定位、翻译起始）知之甚少。本研究的长期目标是确定在卵泡周围颗粒细胞快速分化过程中发生的转录后调控的作用，以确定转录和转录后过程如何介导排卵和黄体生成。该提案将“建立一种方法”来识别基因转录本和生物化学途径，在体内经历独特的转录后调控，并将研究特异性核糖核蛋白（RNP）在排卵中的作用。目的1将通过对多体RNA的表达分析来描述与细胞翻译机制相关的基因（即蛋白质合成的间接描述）。目的2将展示使用特异性RNPs抗体分离和鉴定与类似RNPs相互作用的基因转录本。由于RNP协调转录后的过程，并为细胞内的mrna提供地址（定位），因此，通过与特定RNP关联的独特基因转录物的联系，应该允许我们在新的空间和时间背景下，就转录后调控的基本方面提出有根据的假设。拟议的研究是创新的，在使用翻译（多体RNA）分析方法来填补基因知识的空白，指导排卵过程和黄体化下游的关键内分泌激素，LH。这些使用小鼠模型的体内实验将提供迄今为止哺乳动物系统中激素调节的转录后机制的首次描述。避孕和不育是育龄妇女面临的两大生活质量选择。来自成熟卵泡的可受精卵母细胞的排卵是自然或辅助生殖策略的基础，也是许多不孕妇女中断的关键过程。抑制排卵也是避孕技术的主要目标。这些研究将大大提高我们对黄体生成素如何调节排卵过程的认识。作为了解卵巢功能的基础贡献，这项研究的长期益处可能最终影响更有效的避孕方法和不孕妇女治疗策略的发展。

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.