The translational program in oocyte and early embryo development

卵母细胞和早期胚胎发育的转化程序

基本信息

批准号：
8690104
负责人：
Marco Conti
金额：
$ 34.76万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-15 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8690104
关键词：
Ablation Amphibia Beryllium Binding Proteins Biochemical Bioinformatics Cell Cycle Cell Cycle Regulation Cell Differentiation process Code Data Data Set Databases Defect Development Developmental Process Diagnosis Down-Regulation Embryo Embryonic Development Epigenetic Process Event Failure Female Fertilization Fractionation Genes Genetic Transcription Genetic Translation Genome Germ Lines Health Hereditary Disease Human Human Development Injection of therapeutic agent Invertebrates Maintenance Mammals Maternal Messenger RNA Mediating Meiosis Messenger RNA Microarray Analysis Mining Mission Molecular Profiling Oligonucleotides Oocytes Pattern Phenotype Play Pluripotent Stem Cells Polyribosomes Process Property Proteins Publishing RNA RNA-Binding Proteins Recruitment Activity Regulation Repression Research Role Somatic Cell Staging Stem cells Structure Structure of primordial sex cell System Testing Time Transcript Translating Translational Regulation Translations Tubulin United States National Institutes of Health base egg embryonic stem cell genome-wide improved insight male novel oocyte maturation overexpression pluripotency polymerization preimplantation prevent programs protein expression research study tissue regeneration tool zygote

项目摘要

DESCRIPTION (provided by applicant): The oocyte has the remarkable property of directing the embryo development through accumulation of maternal mRNAs. In the absence of transcription, all the critical events associated with fertilization, reprogramming of the male and female pronucleus, and initial rounds of replication are regulated by a program of mRNA translation. Using a genome wide approach of polysome fractionation and microarray analysis, we have generated a blueprint of the pattern of translation at the oocyte/zygote transition. By mining these data sets as well as published data, it is emerging that the oocyte synthesizes the machinery utilized in the zygote early during oocyte maturation. Among the highly regulated transcripts are those coding for RNA binding proteins Dazl and Pumilio2, critical for germ line development and maintenance of pluripotency in invertebrates. The hypothesis tested is that these RNA regulators play a critical function during early embryo development and, as an extension, in the pluripotent state of the embryonic stem cells. Three Specific Aims are proposed to test this hypothesis. With Specific Aim 1, we propose to characterize the mechanism regulating translation at the oocyte to zygote transition, including the Dazl and Pumilio2. In Specific Aim 2 the function of Dazl and Pumilio2 during the early stages of embryo development will be investigated by conditional ablation of these genes or by disrupting their translation by morpholino oligonucleotide injection. The mechanisms of action of these RNA binding proteins will be investigated by defining their targets. Experiments described in Specific Aim 3 will test the hypothesis that translation mediated by these RNA binding proteins is shared by the embryo and by embryonic stem cells. The phenotype of ES cells deficient in these RNA regulators and the network of mRNA targets will be investigated. This project is significant because it will uncover regulations that are of broad application to different developmental systems. Understanding the regulatory processes of preimplantation embryonic development will reveal new paradigms in epigenetic regulation and reprogramming that will improve current strategies of induction of pluripotent stem cells from somatic cells.

描述（由申请人提供）：卵母细胞具有通过母体mRNA的积累来指导胚胎发育的显着特性。在没有转录的情况下，所有与受精、雄性和雌性原核重新编程以及最初几轮复制相关的关键事件都由 mRNA 翻译程序调节。使用多核糖体分离和微阵列分析的全基因组方法，我们生成了卵母细胞/受精卵转变翻译模式的蓝图。通过挖掘这些数据集以及已发表的数据，人们发现卵母细胞在卵母细胞成熟早期合成受精卵所使用的机制。高度调控的转录物包括那些编码RNA结合蛋白Dazl和Pumilio2的转录物，它们对于无脊椎动物种系发育和多能性的维持至关重要。所测试的假设是，这些 RNA 调节因子在早期胚胎发育过程中发挥着关键作用，并且作为延伸，在胚胎干细胞的多能状态中发挥着关键作用。提出了三个具体目标来检验这一假设。根据具体目标 1，我们建议描述调节卵母细胞到受精卵转变翻译的机制，包括 Dazl 和 Pumilio2。在具体目标2中，将通过条件性消融这些基因或通过注射吗啉代寡核苷酸破坏其翻译来研究Daz1和Pumilio2在胚胎发育早期阶段的功能。这些 RNA 结合蛋白的作用机制将通过定义它们的靶点来研究。具体目标 3 中描述的实验将验证胚胎和胚胎干细胞共享这些 RNA 结合蛋白介导的翻译这一假设。将研究缺乏这些 RNA 调节因子的 ES 细胞的表型和 mRNA 靶标网络。该项目意义重大，因为它将揭示广泛应用于不同发育系统的法规。了解植入前胚胎发育的调控过程将揭示表观遗传调控和重编程的新范例，这将改进当前从体细胞诱导多能干细胞的策略。