CHARACTERIZATION OF ZYGOTE ARREST ONE (ZAR1) IN NONHUMAN PRIMATES

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 Zygote Arrest One（ZAR 1）是小鼠早期胚胎发生所需的关键卵母细胞因子。 Zar 1基因缺失的雌性小鼠由于受精卵阶段的发育停滞而不育，尽管卵泡发生和排卵相当正常。 关于ZAR 1的功能机制以及ZAR 1在小鼠以外的物种中的作用知之甚少。 我们在恒河猴中发现了一个ZAR 1的直系同源基因。 与小鼠Zar 1基因相似，猴ZAR 1优先在卵母细胞和早期植入前胚胎中表达。 我假设ZAR 1是小鼠和恒河猴完全受精和胚胎基因组激活所必需的。 应用体外mRNA注射和RNA干扰技术，在这两个物种的卵母细胞/受精卵中，我提出：（1）拯救来自Zar 1基因敲除小鼠的排卵期卵母细胞和受精卵的发育停滞表型。 目的是确定需要ZAR 1的关键时间，并通过延时活细胞显微镜观察卵母细胞向胚胎转变过程中每个发育阶段的进展和ZAR 1的动态易位;（2）耗尽猕猴完全生长和成熟卵母细胞中的ZAR 1，以证明ZAR 1在植入前胚胎发育中的作用。 目的是阐明ZAR 1在灵长类动物中是否功能保守。 从该项目获得的数据将使我们能够申请R 01基金，用于未来ZAR 1的深入机制研究以及卵母细胞质量评估和女性不孕症治疗的转化研究。

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.