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。 ZAR1是新近发现的一种在卵母细胞中表达的蛋白，在进化过程中保守。缺乏ZAR1的雌性小鼠由于受精卵阶段的胚胎发育受阻而不育。因此，ZAR1属于小鼠卵母细胞向胚胎转化所需的一小部分母体效应蛋白。然而，在人类身上，ZAR1在卵巢和睾丸中都能检测到。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作为我们的哺乳动物系统，因为它们在发育过程中与人类非常相似。该项目的目的是证明ZAR1在早期胚胎发育中的潜在功能，并确定在猴子卵母细胞到胚胎转变过程中发挥作用的关键分子。我们发现了一个与小鼠和人ZAR1高度同源的猴基因，并确定了猴ZAR1基因的表达模式。具体目的如下：(1)研究ZAR1蛋白在猕猴体内的表达。将产生识别ZAR1蛋白高度保守区域的特定抗体，并用于确定ZAR1在猴子和人类中的表达；(2)展示ZAR1在猴子胚胎早期发育中的潜在功能。在体外，RNAi和转基因技术将被用于猴子卵母细胞和早期胚胎；(3)识别在非人类灵长类动物卵母细胞到胚胎转变过程中起作用的关键分子。我们将进行微阵列分析，以确定ZAR1基因敲除胚胎中差异表达的分子。这项研究是首次尝试阐明调控非人类灵长类植入前胚胎发生的分子机制之一。

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 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, 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 monkey embryo development. 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.