MECHANISMS GOVERNING THE OOCYTE-TO-EMBRYO TRANSITION

卵母细胞到胚胎转变的调控机制

• 批准号: 7555698
• 负责人: JOHN J EPPIG
• 金额: $ 25.34万
• 依托单位: JACKSON LABORATORY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-01 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7555698
• 关键词: Affect; Alleles; Bromodomain; Cells; Competence; Development; Doctor of Philosophy; Embryo; Embryonic Development; Exhibits; Female; Female infertility; Fertility; Fertilization; Genes; Genetic; Genomics; Gonadotropins; Growth; Growth and Development function; In Vitro; Infertility; Insemination; Knock-out; Maturation-Promoting Factor; Meiosis; Metaphase; Microarray Analysis; Modeling; Mutation; Names; Nuclear; Oocytes; Pathway interactions; Phenotype; Process; Proteins; Protocols documentation; Public Health; Regulation; Sperm Penetration; Staging; Syndrome; System; Testing; Transcript; Vesicle; WD Repeat; Work; blastocyst; design; egg; follow-up; human female; in vivo; insight; male; mutant; novel; offspring; oocyte maturation; preimplantation; programs; reproductive; research study; sperm cell; zygote

Project III:Mechanisms Governing the Oocyte-to-Embryo Transition John J. Eppig, Project Leader The primary objective of oocyte development is to become an embryo, one capable of development to healthy offspring. Successful oocyte-to-embryo transition depends upon the completion of meiosis and the utilization of maternal factors stored during oocyte development. This ReproGenomics Program Project has produced three infertile mutants that are unable to complete this transition and, therefore, present excellent opportunities to discover novel mechanisms that control it. Meiotic arrest 1 (marfl) mutant oocytes are unable to undergo germinal vesicle breakdown (GVB) and are ovulated at the immature (GV) stage. Oocyte maturation defective (omdl) and bromodomain and WD repeat domain containing 1 (Brwdl) mutant eggs cannot develop beyond the 2 pronuclear stage after insemination. The marfl and omdl mutations affect only females, while the Brwdl mutation affects both males and females. Aim 1 is to discover new pathways regulating the GV-to-GVB transition. The gene harboring the marfl mutation does not encode a component of maturation promoting factor (MPF), or any of its known proximal regulators. Hypotheses that MARF1 protein is either a previously unknown regulator of MPF activity or a key oocyte- specific component of pathways regulated by MPF will be tested. In Aim 2, ovulated GV-arrested marfl mutant oocytes, and oocyte culture systems, will be used to test the hypothesis that some aspects of cytoplasmic maturation, particularly the degradation of specific groups of transcripts during the GV-to-MII transition, and processes that prepare for egg activation, occur independently of nuclear maturation. Aim 3 is to discover new pathways regulating the oocyte-to-embryo transition using the Brwdl and omdl mutations, and a knockout of the oocyte-specific gene zygotic arrest 1 (Zar1) gene. The hypothesis that BRWD1, ZAR1, and OMD1 proteins are fundamental determinants of oocyte quality and that mutant alleles of genes encoding them disrupt key pathways needed to transit from GV-stage oocyte to preimplantation embryo will be tested. Relevance to Public Health: By discovering novel mechanisms controlling the oocyte-to-embryo transition, this work will provide insight into common human female infertility syndromes. The studies proposed here will discover molecules, pathways, and processes needed to produce "good" eggs capable of maturation and undergoing early embryogenesis.

项目三：卵母细胞向胚胎转变的调控机制 约翰·J·埃皮格 (John J. Eppig)，项目负责人 卵母细胞发育的主要目标是成为胚胎，能够发育成胚胎 健康的后代。卵母细胞向胚胎的成功转变取决于减数分裂的完成和 利用卵母细胞发育过程中储存的母体因子。该 ReproGenomics 计划项目有 产生了三个不育突变体，它们无法完成这一转变，因此表现出优异的 发现控制它的新机制的机会。减数分裂停滞 1 (marfl) 突变卵母细胞是 无法经历生发囊泡破裂 (GVB)，并在未成熟 (GV) 阶段排卵。 卵母细胞成熟缺陷 (omdl) 和含溴结构域和 WD 重复结构域 1 (Brwdl) 受精后突变卵不能发育超过第二个原核阶段。 marfl 和 omdl 突变仅影响女性，而 Brwdl 突变影响男性和女性。目标 1 是 发现调节 GV 到 GVB 转变的新途径。携带 marfl 突变的基因确实 不编码成熟促进因子 (MPF) 或其任何已知的近端调节因子的成分。 假设 MARF1 蛋白要么是以前未知的 MPF 活性调节剂，要么是关键的卵母细胞 将测试 MPF 调节的途径的特定组成部分。在目标 2 中，排卵 GV 阻滞 marfl 突变卵母细胞和卵母细胞培养系统将用于检验以下假设： 细胞质成熟，特别是 GV 至 MII 期间特定转录物组的降解 转变和准备卵子激活的过程独立于核成熟而发生。目标 3 是利用 Brwdl 和 omdl 发现调节卵母细胞到胚胎转变的新途径 突变，以及卵母细胞特异性基因合子停滞 1 (Zar1) 基因的敲除。假设 BRWD1、ZAR1 和 OMD1 蛋白是卵母细胞质量的基本决定因素，突变等位基因 编码它们的基因破坏了从 GV 期卵母细胞过渡到植入前所需的关键途径 胚胎将接受测试。 与公共卫生的相关性：通过发现控制卵母细胞到胚胎的新机制 转变，这项工作将深入了解常见的人类女性不孕综合症。研究 这里提出的将发现产生“好”鸡蛋所需的分子、途径和过程 成熟并经历早期胚胎发生。