IDENTIFICATION/CHARACTERIZATION OF NOVEL MATERNAL-EFFECT GENE PRODUCTS

点击翻译按钮获取中文摘要

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 gene products direct a series of crucial events occurring at the oocyte-to-embryo transition, including the completion of the first meiosis, the initiation of the second meiosis, and the activation of the embryo genome. Only a few of these gene products, termed maternal effect proteins, have been identified and little is known about the mechanisms mediating their functions. We identified several novel potential maternal effect genes by in silico subtraction. After initial gene expression analysis in mice and macaques, we decided to focus on a newly described protein family--NLR family, pyrin domain containing protein (NLRP). The function of the majority members in this family is unknown. However, several Nlrp genes are found exclusively in germ cells in rodents; and at least one of them, NLRP5, is essential to early embryo development in mice. Based on previous findings in mice and our preliminary data in macaques, we hypothesize that oocyte-specific NLRPs are required for oocyte maturation, fertilization, and early embryo development; thus, knocking down one or more NLRP genes in the macaque oocyte will impair the progress of oocyte-to-embryo transition, resulting in unfertilizable eggs or developmental arrest of preimplantation embryos. To further characterize oocyte-specific NLRPs in macaques, we propose to demonstrate the spatiotemporal expression of NLRP proteins and perform functional studies using in vitro RNA interference (RNAi) in the oocyte. In addition, we will generate Nlrp gene knockout/knockdown mouse models to explore their in vivo functions in rodents.