IDENTIFICATION/CHARACTERIZATION OF POTENTIAL NOVEL MATERNAL-EFFECT GENE PRODUCTS

潜在新型母体效应基因产品的鉴定/表征

• 批准号: 8173203
• 负责人: Xuemei Wu
• 金额: $ 4.76万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8173203
• 关键词: Cells; Computer Retrieval of Information on Scientific Projects Database; Data; Domestic Animals; Embryo; Embryonic Development; Experimental Designs; Factor Analysis; Family; Family member; Funding; Generations; Genes; Grant; Human; In Vitro; Institution; Knockout Mice; Leucine-Rich Repeat; Link; Macaca; Macaca mulatta; Mammals; Mediating; Monkeys; Mus; Mutation; Oocytes; Oogenesis; Play; Proteins; RNA Interference; Research; Research Personnel; Resources; Rodent; Role; Source; Staging; United States National Institutes of Health; blastocyst; in vivo; marenostrin; mouse model; nonhuman primate; novel; protein protein interaction; reproductive; yeast two hybrid system

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We have identified a group of six NLRP (NLR family, pyrin domain containing) genes that are specifically expressed in the oocyte and early preimplantation embryos in rhesus macaques. Mouse NLRP5 has been demonstrated to be essential for embryogenesis beyond the 2-cell stage, and human NLRP7 was identified as the first maternal effect gene as the mutations of which have been linked to various forms of reproductive wastage. However, functions of other NLRP family members are not known. Our preliminary data in nonhuman primates, along with recent findings in rodents and domestic animals, suggest that these oocyte-specific NLRP genes play important roles during oogenesis and early embryo development in mammals. The purpose of this project is to reveal the functions of selected oocyte-specific NLRPs in mice and rhesus macaques. The specific aims are to: (1) characterize in vivo functions of mouse NLRP4A and 9B using knockout mouse models; (2) demonstrate the in vitro postovulatory roles of selected NLRP genes (mouse Nlrp4a, 5 and 9b; macaque NLRP4, 5 and 9) in mouse and macaque oocytes; (3) identify and analyze factors that interact with selected NLRP proteins in mouse and macaque oocytes. NLRP proteins contain a pyrin domain (PYD), a NACHT, and a leucine rich repeat (LRR) domain; both the PYD and LRR mediate protein-protein interactions. Experimental designs include generation and analysis of gene knockout mouse models, RNA interference (RNAi) in mouse and monkey oocytes, and application of yeast two-hybrid system in the mouse and macaque oocyte. We have demonstrated that NLRP5 is required for the oocyte-to-embryo transition in nonhuman primates. Functional studies with Nlrps 4a and 9b in mice, and NLRPs 4 and 9 in monkeys are still ongoing.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 我们已经鉴定了一组六个 NLRP（NLR 家族，含有热蛋白结构域）基因，这些基因在恒河猴的卵母细胞和早期植入前胚胎中特异性表达。 小鼠 NLRP5 已被证明对于 2 细胞阶段之后的胚胎发生至关重要，而人类 NLRP7 被确定为第一个母体效应基因，因为其突变与多种 生殖浪费的形式。 然而，NLRP 家族其他成员的功能尚不清楚。 我们在非人类灵长类动物中的初步数据，以及最近在啮齿动物和家畜中的发现表明，这些卵母细胞特异性 NLRP 基因在哺乳动物的卵子发生和早期胚胎发育过程中发挥着重要作用。 该项目的目的是揭示小鼠和恒河猴中选定的卵母细胞特异性 NLRP 的功能。 具体目标是：（1）利用敲除小鼠模型表征小鼠NLRP4A和9B的体内功能； (2) 证明选定的 NLRP 基因（小鼠 Nlrp4a、5 和 9b；猕猴 NLRP4、5 和 9）在小鼠和猕猴卵母细胞中的体外排卵后作用； (3) 鉴定并分析与小鼠和猕猴卵母细胞中选定的 NLRP 蛋白相互作用的因素。 NLRP 蛋白包含一个热蛋白结构域 (PYD)、一个 NACHT 和一个富含亮氨酸重复序列 (LRR) 结构域； PYD 和 LRR 均介导蛋白质-蛋白质相互作用。 实验设计包括基因敲除小鼠模型的生成和分析、小鼠和猴卵母细胞中的RNA干扰（RNAi）以及酵母双杂交系统在小鼠和猕猴卵母细胞中的应用。我们已经证明 NLRP5 是非人灵长类动物卵母细胞到胚胎转变所必需的。 Nlrps 4a 和 9b 在小鼠中的功能研究以及 NLRPs 4 和 9 在猴子中的功能研究仍在进行中。