Gene Expression in the Preimplantation Mouse Embryo

植入前小鼠胚胎中的基因表达

• 批准号: 7936524
• 负责人: RICHARD M SCHULTZ
• 金额: $ 29.28万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2011-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7936524
• 关键词: Assisted Reproductive Technology; Attention; Binding; Binding Proteins; Candidate Disease Gene; Cell physiology; Cells; Chromatin; Chromatin Structure; Competence; DNA; Development; Embryo; Embryonic Development; Enzymes; Family; Fertilization; Fostering; Funding; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Genome; Goals; Grant; Growth; Histone H3; Histones; Human; Human Development; Infertility; Knowledge; Link; Mediating; Meiosis; MicroRNAs; Molecular; Morula; Mus; Oocytes; Participant; Pattern; Phase; Post-Transcriptional Regulation; Pre-implantation Embryo Development; Process; Prophase; RNA Interference; Recruitment Activity; Research Personnel; Role; Series; Staging; Testing; Transcript; Transferase; Transgenic Organisms; Ubiquitin; blastocyst; egg; embryo stage 2; helper-dependent adenoviral vector; heterochromatin-specific nonhistone chromosomal protein HP-1; improved; mRNA Transcript Degradation; member; oocyte maturation; preimplantation; programs; research study; therapy development; transcription factor; zygote

DESCRIPTION (provided by applicant): The long-term goal of this application is to determine underlying mechanisms that govern changes in gene expression during oocyte growth and the maternal-to-zygotic transition, both of which are linked and essential for development. A universal feature of oocyte development is that transcription declines commencing around mid-growth such that fully-grown oocytes are essentially transcriptionally quiescent. In mouse, this decline correlates with visible changes in chromatin structure, which becomes more condensed and decreased activity of the transcription machinery. Using a transgenic RNAi approach that permits study of the function of any gene in oocyte development, Specific Aim 1 will test the hypothesis that members of the HP1 family of chromatin-binding proteins and UBC9, via its central role in sumoylation, are essential for changes in chromatin structure and transcriptional activity that occur during oocyte growth. A major reprogramming of gene expression is first detected during the 2-cell stage and essential for continued development. Superimposed on genome activation is development of a chromatin-mediated transcriptionally repressive state. Our transcript profiling experiments identified Myc as a candidate gene pivotal for genome activation and reprogramming of gene expression, and Hdad as essential for establishing the transcriptionally repressive state. Using an RNAi approach to target Myc and Hdad, Specific Aim 2 will test the hypothesis that Myc and Hdad are critical for these two processes that collaboratively sculpt the appropriate pattern of gene expression required for successful development following genome activation. Oocytes express miRNAs and mRNA degradation, which initiates during oocyte maturation and continues during early preimplantation development, is a post-transcriptional mechanism that contributes to determining the global pattern of gene expression in the embryo. Specific Aim 3 will test the hypothesis that miRNAs target specific mRNAs for degradation in P-bodies and that this mechanism is essential for proper oocyte/embryo development. The proposed studies will increase our basic knowledge and understanding of human development. In the near term, they may help improve treatment of human infertility by providing new knowledge that will facilitate the rational development of treatments that foster improved oocyte and preimplantation embryo development in the practice of Assisted Reproductive Technology (ART).

描述（由申请人提供）：本申请的长期目标是确定卵母细胞生长过程中基因表达变化的基本机制和孕妇到二旋转过渡的变化，这两者都链接并且对于发育至关重要。卵母细胞发育的一个普遍特征是转录下降开始于中期生长，因此完全生长的卵母细胞本质上是转录静止的。在小鼠中，这种下降与染色质结构的可见变化相关，后者变得更加凝结并减少了转录机械的活性。使用允许研究任何基因在卵母细胞发育中的功能的转基因RNAI方法，具体目标1将检验以下假设：染色质结合蛋白和UBC9的HP1家族的成员通过其在Sumoylation中的核心作用，对于在卵母细胞生长过程中发生的染色质结构和转录活性的变化至关重要。首先在2细胞阶段检测到基因表达的主要重编程，对于持续发展至关重要。基因组激活叠加的是染色质介导的转录抑制状态的发展。我们的转录分析实验将MYC鉴定为基因组激活和重编程基因表达的候选基因关键，而HDAD对于建立转录抑制状态至关重要。使用RNAi方法靶向MYC和HDAD，具体目标2将检验以下假设：MYC和HDAD对于这两个过程至关重要，这些过程与基因组激活后成功开发所需的合适基因表达模式至关重要。卵母细胞表达miRNA和mRNA降解，在卵母细胞成熟过程中启动，并在早期植入前发育过程中继续进行，是一种转录后机制，有助于确定胚胎中基因表达的全局模式。具体目标3将检验以下假设：miRNA靶向特定的mRNA用于p-bodies中的降解，并且该机制对于适当的卵母细胞/胚胎发育至关重要。拟议的研究将增加我们对人类发展的基本知识和理解。在短期内，它们可以通过提供新知识来帮助改善人类不育症的治疗，从而有助于促进疗法的疗法促进卵母细胞和植入前胚胎发展的理性发展，从而促进辅助生殖技术（ART）。