IDENTIFICATION/CHARACTERIZATION OF POTENTIAL NOVEL MATERNAL-EFFECT GENE PRODUCTS

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来源获得了主要资金，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。卵母细胞衍生的母体效应蛋白指导卵母细胞向胚胎转变时发生的一系列关键事件，包括第二次减数分裂的完成、第一次有丝分裂的开始和胚胎基因组的激活。很少有母效应蛋白已被确定，很少有人知道的机制，介导其功能。这项研究的目的是确定和表征新的潜在的母效应基因。利用计算机差减技术，我们在小鼠中鉴定了四个新基因，命名为xw 13，xw 44，xw 46和xw 47。基于这些mRNA的空间和时间分布，我假设这四个新基因编码小鼠卵母细胞向胚胎过渡所需的母体效应蛋白。设计了两个具体目标：目标1，表征由这些新基因编码的基因产物。我们将产生特异性抗体对四个新的推导蛋白质，以表征其蛋白质的结构和表达在小鼠组织和早期胚胎，并确定其直系同源物在非人灵长类动物和人类;目的2，阐明新的基因产物在卵母细胞到胚胎的过渡在体外的功能意义。在小鼠卵母细胞和早期胚胎中应用体外RNAi和转基因方法，我们将研究新基因产物在卵母细胞成熟，受精和早期胚胎发生过程中的潜在功能。大约50%的妊娠丢失与人类胚胎植入前缺陷有关。因此，母体效应蛋白的研究将提供关键的见解，可能的基因产物，涉及在特发性人类不育症。

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. Oocyte-derived maternal-effect proteins direct a series of crucial events occurring at the oocyte-to-embryo transition, including the completion of the second meiosis, the initiation of the first mitosis, and the activation of the embryonic genome. Few maternal-effect proteins have been identified and little is known about the mechanisms mediating their functions. The goal of this research is to identify and characterize novel potential maternal-effect genes. Using in silico subtraction, we have identified four novel genes named xw13, xw44, xw46 and xw47 in mice. Based on the spatial and temporal distribution of these mRNAs, I hypothesize that the four novel genes encode maternal effect proteins that are required for the oocyte-to-embryo transition in mice. Two specific aims are designed: Aim 1, characterization of gene products encoded by these novel genes. We will generate specific antibodies against the four novel deduced proteins to characterize their protein structure and expression in mouse tissues and early embryos, and identify their orthologs in nonhuman primates and humans; Aim 2, elucidation of the functional significance of the novel gene products in the oocyte-to-embryo transition in vitro. Applying in vitro RNAi and transgene methodologies in mouse oocytes and early embryos, we will study the potential functions of the novel gene products during oocyte maturation, fertilization, and early embryogenesis. Approximately 50% of pregnancy loss is associated with preimplantation defects in humans. Thus, the studies on maternal effect proteins will provide key insights into possible gene products that are implicated in idiopathic human infertility.