Mechanisms Controlling Oocyte Developmental Competence and Embryo

控制卵母细胞发育能力和胚胎的机制

• 批准号: 8286510
• 负责人: Marco Conti
• 金额: $ 32.55万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-01 至 2017-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8286510
• 关键词: Affect; Amphiregulin; Animal Model; Assisted Reproductive Technology; Biological; Biological Assay; Biological Markers; Cell Communication; Cell Cycle; Chromatin Remodeling Factor; Code; Collaborations; Competence; Data; Data Analyses; Development; Diagnosis; EGF gene; Embryo; Embryonic Development; Environment; Failure; Female infertility; Fertilization; Fertilization in Vitro; Genetic Models; Genetic Transcription; Genetic Translation; Genome; Germ Cells; Human; In Vitro; Infertility; Instruction; Knockout Mice; Laboratories; Lead; Maternal Messenger RNA; Measurement; Meiosis; Messenger RNA; Methods; Modeling; Molecular; Monitor; Mus; Nature; Oocytes; Patients; Pattern; Play; Process; Protein Secretion; Proteins; Regulation; Reporter; Research; Resolution; Rodent; Role; Scientist; Signal Transduction; Somatic Cell; Students; Testing; Time; Transcript; Translating; Translational Regulation; Translations; Work; assisted reproduction; base; chromatin remodeling; clinical practice; endometriosis; genome-wide; genome-wide analysis; global environment; high school; human female; improved; in vitro Model; in vivo; nuclear reprogramming; oocyte maturation; programs; research study; tool; zygote

PROJECT SUMMARY (See instructions): PROJECT I The overarching hypothesis that we will test with this proposal is that human female infertility and assisted reproductive technology (ART) failures are, in part, caused by a disruption of the maternal mRNA translational program. Using a genome-wide strategy, we have assembled a blueprint of the translational regulations active during mouse oocyte maturation. We demonstrated that a large number of mRNAs coding for cell cycle components and for components of the transcriptional and chromatin remodeling machinery are translated following a well defined succession early during oocyte maturation. These findings have led to the hypothesis that timed translation of a subset of maternal mRNAs is critical for the oocyte to develop as an embryo. Moreover, we propose that somatic cell signals control this translational program of the oocyte. On the basis of preliminary data showing that the EGF-network plays a role in these somatic-germ cell interactions, we will explore how these signals contribute to developmental competence. The experimental plan is organized along three specific aims. With the first Aim, the oocyte translation program will be characterized in in vivo genetic models of compromised competence or after in vitro maturation. In the second Aim, the mechanisms by which somatic cells regulate translation in the oocytes will be investigated using in vitro models where somatic germ cell interactions are maintained and with reporters monitoring translation of candidate transcripts. The third Aim will focus on translation of mRNAs coding for oocyte secretory products to predict actual protein secretion. These measurements will be used as a readout of correct execution of the translational program. Secretion from human oocytes will be used for proof of principle that these patterns reflect the competence of the oocyte to sustain embryo development. The concepts developed with this project will open new avenues for monitoring oocyte quality in assisted reproduction in clinical practice.

项目总结（见说明）：项目I 我们将用这一提议测试的总体假设是，人类女性不孕症和辅助生殖技术（ART）失败部分是由母体mRNA翻译程序的中断引起的。使用全基因组策略，我们已经组装了小鼠卵母细胞成熟过程中活跃的翻译调控的蓝图。我们证明了大量的mRNA编码 对于细胞周期成分和转录和染色质重塑机制的成分，在卵母细胞成熟早期遵循明确的连续性进行翻译。这些发现提出了这样的假设：母体mRNA子集的定时翻译对于卵母细胞发育为胚胎至关重要。此外，我们提出，体细胞信号控制这个翻译程序的卵母细胞。对 初步数据显示，EGF网络在这些体细胞-生殖细胞相互作用中发挥作用的基础上，我们将探讨这些信号如何促进发育能力。实验计划是按照沿着三个具体目标组织的。对于第一个目标，卵母细胞翻译程序将在能力受损的体内遗传模型中或在体外成熟后进行表征。在第二个目标中，体细胞调节卵母细胞翻译的机制将使用体外模型进行研究，在体外模型中体细胞生殖细胞的相互作用得到维持，并与报告监测候选转录本的翻译。第三个目标将集中在翻译的mRNA编码的卵母细胞分泌产物，以预测实际的蛋白质分泌。这些测量值将用作正确执行平移程序的读数。人卵母细胞的分泌物将用于证明这些模式反映卵母细胞维持胚胎发育的能力的原理。该项目开发的概念将为临床实践中辅助生殖中监测卵母细胞质量开辟新的途径。