CHARACTERIZATION OF ZYGOTE ARREST ONE (ZAR1) IN NONHUMAN PRIMATES

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。ZAR 1是一个新发现的在进化过程中保守的卵母细胞表达蛋白。缺乏ZAR 1的雌性小鼠由于受精卵阶段胚胎发生受阻而不育。因此，ZAR 1属于小鼠卵母细胞向胚胎转变所需的一小组母体效应蛋白。然而，在人类中，ZAR 1 mRNA在卵巢和睾丸中都可检测到。为了阐明ZAR 1在人类生育中的功能，我们选择非人灵长类动物作为我们的哺乳动物系统，因为它们在发育方面与人类非常相似。该项目的目的是证明ZAR 1在早期胚胎发育过程中的潜在功能，并确定在猴卵母细胞向胚胎过渡中发挥作用的关键分子。我们发现了一个与小鼠和人ZAR 1基因高度同源的猴cDNA，并确定了猴ZAR 1 mRNA的表达模式。具体目的如下：（1）表征ZAR 1蛋白在猴中的表达。将产生识别ZAR 1蛋白的高度保守区域的特异性抗体，并用于确定ZAR 1在猴和人中的表达;（2）证明ZAR 1在猴早期胚胎发育期间的潜在功能。将在猴卵母细胞和早期胚胎中采用体外RNAi和转基因技术;（3）鉴定在非人灵长类动物卵母细胞向胚胎转变中起作用的关键分子。我们将进行微阵列分析，以确定差异表达的分子在ZAR 1敲低胚胎。这里描述的研究是第一次试图说明调节非人类灵长类动物植入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.