ROLE OF THE ANTIZYME FAMILY DURING XENOPUS DEVELOPMENT

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。多胺是重要的有机阳离子，对生命至关重要。活跃增殖的细胞，如胚胎细胞或癌细胞，对多胺有严格的需求。因此，多胺水平的操纵可以对生长产生深远的影响。细胞维持良好调节的途径，将多胺水平控制在细胞生长的最佳水平。抗酶（AZ）是一种重要的蛋白质，通过控制多胺的水平来影响细胞增殖的速率。本项目的目的是表征多胺水平的调节和对细胞生长的影响，使用的发展模型的增殖，非洲爪蟾胚胎发生。这项工作的重点将是确定AZ如何调节非洲爪蟾胚胎细胞的增殖能力。目前还没有一个系统的研究抗酶的功能，使用的发展模式系统，如两栖动物非洲爪蟾。本项目将利用定量RT-PCR、Northern、Westerns和二维蛋白质凝胶技术，对非洲爪蟾胚胎早期发育过程中AZ基因家族的表达模式进行表征。AZ家族的功能将通过显微注射实验来研究，该实验将迫使AZ在单细胞胚胎阶段表达。这将允许AZ是否可以显性控制细胞增殖的研究。因此，该项目的结果将是AZ家族在使用易处理的生物体非洲爪蟾的增殖和发育中的作用的阐明。由于非洲爪蟾的发育途径与相同的哺乳动物途径密切相关，因此该项目将有助于阐明AZ在小鼠和人类中的作用，并可能应用于癌细胞的生长。

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. Polyamines are important organic cations that are essential for life. Cells that are actively proliferating such as embryonic cells or cancer cells, have strict needs for polyamines. Thus, manipulation of polyamine levels can have profound effects on growth. Cells maintain well-regulated pathways that control polyamine levels at optimum levels for cell growth. Antizyme (AZ) is a crucial protein that influences the rate of cell proliferation by controlling the levels of polyamines. An aim of this project is to characterize the regulation of polyamine levels and effects on cell growth using a developmental model of proliferation, Xenopus laevis embryogenesis. A focus of the work will be to determine how AZ regulates the proliferative capacity of Xenopus embryonic cells. There has not been a systematic study of antizyme function using a developmental model system such as the amphibian Xenopus laevis. This project will characterize the expression pattern of the AZ gene family using quantitative RT-PCR, Northerns, Westerns, and two dimensional protein gels during the early development of Xenopus embryos. The function of the AZ family will be studied by microinjection experiments that will force AZ expression at the one cell embryo stage. This will allow for a study of whether AZ can dominantly control cell proliferation. Thus, the outcome of this project will be the elucidation of the role of the AZ family in proliferation and development using a tractable organism, Xenopus laevis. Since the developmental pathways of Xenopus laevis are closely related to the same mammalian pathways, this project will help to shed light on the role of AZ in mice and humans with potential applications to the growth of cancer cells.