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III：管理卵母细胞到embryo过渡的机制 项目负责人约翰·J·埃皮格（John J. Eppig） 卵母细胞开发的主要目标是成为胚胎，一个能够发育的胚胎 健康的后代。成功的卵母细胞到永远的过渡取决于减数分裂和 在卵母细胞发育过程中储存的母体因素的利用。这个重生程序项目有 产生了三个无法完成此过渡的不育突变体 发现控制它的新型机制的机会。减数分裂逮捕1（MARFL）突变卵母细胞是 无法经历生发囊泡分解（GVB），并且在未成熟（GV）阶段排卵。 卵母细胞成熟有缺陷（OMDL）和溴结构域和WD重复域，含有1（BRWDL） 突变卵不能在接送后两个前核阶段出现。 Marfl和OMDL 突变仅影响女性，而BRWDL突变会影响男性和女性。目标1是 发现调节GV到GVB过渡的新途径。携带MARFL突变的基因确实 不编码成熟促进因子（MPF）或其任何已知近端调节剂的组成部分。 假设MARF1蛋白是以前未知的MPF活性调节剂，或者是关键的卵母细胞 将测试由MPF调节的途径的特定组成部分。在AIM 2中，排卵的GV捕捞 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，尤其是在GV-TO-MII期间特定转录本的降解 过渡和准备卵子激活的过程独立于核成熟。目标3 是发现使用BRWDL和OMDL调节卵母细胞到embryo过渡的新途径 突变，以及卵母细胞特异性基因性滞留1（ZAR1）基因的敲除。假设是 BRWD1，ZAR1和OMD1蛋白是卵母细胞质量的基本决定因素和突变等位基因 编码它们的基因破坏从GV阶段卵母细胞过渡到植入术所需的关键途径 将测试胚胎。 与公共卫生的相关性：通过发现控制卵母细胞到embryo的新型机制 过渡，这项工作将洞悉普通的人类女性不育症综合症。研究 这里提出的将发现产生能够“好”卵所需的分子，途径和过程 成熟并经历早期胚胎发生。