CHARACTERIZATION OF ZYGOTE ARREST ONE (ZAR1) IN NONHUMAN PRIMATES

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 受精卵抑制因子1(ZAR1)是小鼠早期胚胎发育所必需的关键卵母细胞因子。Zar1基因缺失的雌性小鼠由于受精卵阶段发育停滞而不育，尽管卵泡发生和排卵相当正常。关于ZAR1的S功能的机制以及ZAR1在除小鼠以外的物种中的作用，目前还知之甚少。我们在恒河猴中发现了一个ZAR1直系同源基因。与小鼠ZAR1基因类似，猴子ZAR1基因在卵母细胞和早期植入前胚胎中优先表达。我推测ZAR1是小鼠和恒河猴完全受精和胚胎基因组激活所必需的。在两种动物的卵母细胞/受精卵中应用体外mRNA注射和RNAi，我建议：(1)挽救Zar1基因缺失小鼠的排卵期卵母细胞和受精卵的发育停滞表型。其目的是确定需要ZAR1的关键时间，并通过时间推移活细胞显微镜观察每个发育阶段的进展和ZAR1在卵到胚胎转变过程中的动态转位；(2)在完全发育和成熟的卵母细胞中去除ZAR1，以证明ZAR1在植入前胚胎发育中的作用。其目的是阐明ZAR1在灵长类动物中是否在功能上保守。从该项目获得的数据将使我们能够申请R01资金，用于未来对ZAR1的深入机制研究，以及女性卵母细胞质量评估和不孕治疗的翻译研究。

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. Zygote Arrest One (ZAR1) is a key oocyte factor required for early embryogenesis in mice. Zar1 null female mice are infertile due to a developmental arrest at the zygote stage despite fairly normal folliculogenesis and ovulation. Little is known about the mechanism underlying ZAR1's functions as well as the roles of ZAR1 in species other than mice. We identified a ZAR1 ortholog in rhesus monkeys. Similar to the mouse Zar1 gene, monkey ZAR1 is preferentially expressed in the oocyte and early preimplantation embryos. I hypothesize that ZAR1 is required for complete fertilization and the embryonic genome activation in both mice and rhesus macaques. Applying in vitro mRNA injection and RNAi in the oocyte/zygote from both species, I propose to: (1) rescue the developmental arrest phenotype in periovulatory oocytes and zygotes derived from Zar1 null mice. The goal of the aim is to determine the critical timing when ZAR1 is required, and to observe the progress of each developmental stage and dynamic translocalization of ZAR1 during the oocyte-to-embryo transition by time-lapse live cell microscopy; and (2), deplete macaque ZAR1 in fully-grown and mature oocytes to demonstrate the role of ZAR1 in preimplantation embryo development. The goal of the aim is to elucidate whether ZAR1 is functionally conserved in primates. Data obtained from this project will allow us to apply for R01 funds for future in-depth mechanistic studies on ZAR1 and translational research in oocyte quality assessment and infertility therapies in women.