Mechanisms Controlling Oocyte Developmental Competence and Embryo

项目总结(见说明)：项目I 我们将用这一提议检验的最重要的假设是，人类女性不孕不育和辅助生殖技术(ART)失败的部分原因是母体mRNA翻译程序的中断。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。我们证明了大量编码mRNAs 对于细胞周期成分以及转录和染色质重塑机制的成分，在卵母细胞成熟的早期，按照明确的顺序进行翻译。这些发现导致了一种假设，即母体mRNAs子集的定时翻译对卵母细胞作为胚胎发育至关重要。此外，我们认为，体细胞信号控制着卵母细胞的这一翻译程序。在……上面 在初步数据显示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，我们将探索这些信号是如何影响发育能力的。这项实验计划按照三个具体目标组织。第一个目标是，卵母细胞翻译计划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，将使用保持体细胞生殖细胞相互作用的体外模型和记者监控候选转录本的翻译来研究体细胞调控卵母细胞翻译的机制。第三个目标将集中在翻译编码卵母细胞分泌产物的mRNAs，以预测实际的蛋白质分泌。这些测量将被用作翻译程序正确执行的读数。人类卵母细胞的分泌物将被用来证明这些模式反映了卵母细胞维持胚胎发育的能力。该项目开发的概念将为临床实践中监测辅助生殖中的卵母细胞质量开辟新的途径。

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.