CHARACTERIZATION OF ZYGOTE ARREST ONE (ZAR1) IN NONHUMAN PRIMATES

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。ZAR1是一种在进化过程中保守的新发现的卵母细胞表达蛋白。缺乏ZAR1的雌性小鼠由于在受精卵阶段胚胎发育受阻而不育。因此，ZAR1属于一小群母系效应蛋白，这些蛋白是小鼠卵细胞向胚胎转变所必需的。然而，在人类中，ZAR1 mRNA在卵巢和睾丸中都可以检测到。为了阐明ZAR1在人类生育中的功能，我们选择了非人类灵长类动物作为我们的哺乳动物系统，因为它们在发育上与人类非常相似。该项目的目的是证明ZAR1在早期胚胎发生中的潜在功能，并确定在猴卵细胞向胚胎转变过程中起作用的关键分子。我们发现了一个与小鼠和人类ZAR1基因高度同源的猴子cDNA，并确定了猴子ZAR1 mRNA的表达模式。具体目的如下：(1)表征ZAR1蛋白在猴子体内的表达。将产生识别ZAR1蛋白高度保守区域的特异性抗体，并用于测定猴子和人类的ZAR1表达；(2)阐明ZAR1在猴子早期胚胎发育中的潜在功能。体外RNAi和转基因技术将用于猴卵母细胞和早期胚胎；(3)鉴定非人灵长类动物卵母细胞向胚胎转变的关键分子。我们将进行微阵列分析，以鉴定ZAR1敲低胚胎中的差异表达分子。这里描述的研究是第一次尝试阐明调节非人类灵长类动物着床前胚胎发生的分子机制。

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. ZAR1 is a newly identified oocyte-expressed protein conserved during evolution. Female mice lacking ZAR1 are infertile due to a block of embryogenesis at the zygote stage. Thus, ZAR1 belongs to a small group of maternal-effect proteins that are required for the oocyte-to-embryo transition in mice. In humans, however, ZAR1 mRNA is detectable in both the ovary and the testis. To elucidate functions of ZAR1 in human fertility, we chose nonhuman primates as our mammalian system due to their close resemblance to humans in development. The aim of this project is to demonstrate potential functions of ZAR1 during early embryogenesis and identify key molecules that function at the oocyte-to-embryo transition in monkeys. We have discovered a monkey cDNA that is highly homologous to mouse and human ZAR1 gene, and determined the expression pattern of monkey ZAR1 mRNA. The specific aims are as follows: (1) to characterize ZAR1 protein expression in monkeys. Specific antibodies that recognize a highly conserved region of ZAR1 protein will be generated and used to determine ZAR1 expression in monkeys and humans; (2) to demonstrate potential functions of ZAR1 during early embryo development in monkeys. In vitro RNAi and transgenic techniques will be employed in monkey oocytes and early embryos; (3) to identify key molecules that function at the oocyte-to-embryo transition in nonhuman primates. We will perform microarray analysis to identify differentially expressed molecules in ZAR1 knockdown embryos. The study described here is among the first attempts to illustrate molecular mechanisms that regulate preimplantation embryogenesis in nonhuman primates.