Research Starter Grant: Analysis of ODS-1 in C. elegans Exposed to Anoxia

研究启动资助：分析暴露于缺氧的线虫中的 ODS-1

• 批准号: 0307491
• 负责人: Pamela Padilla
• 金额: $ 5万
• 依托单位: University of North Texas
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-06-01 至 2004-05-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0307491&HistoricalAwards=false
• 关键词: Research; Starter; Grant; Analysis; ODS

Oxygen deprivation influences the growth and development of organisms. Many organisms in nature are subjected to changes in oxygen levels and have adapted to survive oxygen deprivation. The soil nematode Caenorhabditis elegans is capable of surviving a wide range of oxygen levels and the developmental progression of the organism depends on the oxygen concentration. For example, C. elegans exposed to a hypoxic environment (0.5% to 1.5% oxygen) develop slowly, but nematodes exposed to an anoxic environment (0% oxygen) arrest in their developmental and cell cycle progression. This arrest, anoxia-induced suspended animation, is reversible upon re-exposure to oxygen. Organisms other than C. elegans, such as zebrafish, brine shrimp, and fruit flies are also capable of surviving anoxia by arresting development. Little is known about the developmental and cell cycle arrest in response to anoxia. To gain a full understanding of anoxia-induced suspended animation it is necessary to understand the genetic, physiologic, and cellular responses to anoxia in a variety of metazoans at different stages of development.Embryos exposed to anoxia arrest developmental and cell cycle progression. It is not known if there is a genetic and cellular basis for anoxia induced cell cycle arrest. Using a genetic model system such as C. elegans to study anoxia induced cell cycle arrest will help determine if there is a genetic response to anoxia. C. elegans is a model system in which classical forward and reverse genetic analysis is possible. The large size and transparency of nematode embryos make them an excellent system for observing cell cycle events during early development. Additionally, C. elegans chromosomes are holocentric, with kinetochores that extend the length of the chromosomes. Thus, during anaphase chromosomes move as entire units without lagging arms. Furthermore, the large size of C. elegans kinetochores (up to 4 microns) make it easy to study cell cycle events. Thus, the recent finding by Dr. Padilla that the nematode embryo arrests cell cycle progression in anoxia, make the nematode a useful system for understanding the cellular and genetic responses required for oxygen deprivation survival.Dr. Padilla's long-term research goal is to determine the mechanisms employed by C. elegans to respond to severe oxygen deprivation. The hypothesis of this one-year project is that a gene (ODS-1), identified by an RNA interference (RNAi) screen for genes that are essential for C. elegans embryos to survive anoxia, is a component of the spindle checkpoint in the nematode embryo. The research will focus on the characterization of ODS-1, and consists of the following two aims:Aim 1. To determine the subcellular localization of ODS-1 protein in embryos exposed to a normoxic or anoxic environment.Aim 2. To evaluate the phenotype of ODS-1 (RNAi) embryos.This research will result in the characterization of a gene and gene product that appears to be important for nematode embryos to survive anoxia, and will permit the testing of the hypothesis that spindle checkpoints are involved in anoxia-induced cell cycle arrest.Broader impacts: This is a Research Starter Grant, awarded to an NSF Postdoctoral Fellow who has accepted a tenure-track position at an eligible institution, as described in NSF 00-139. Dr. Padilla actively integrates research and education by recruiting undergraduate and graduate students to work on research projects in her lab, as well as by teaching in the undergraduate classroom.

缺氧会影响生物体的生长和发育。自然界中的许多生物体都受到氧气水平的变化，并适应了缺氧的生存。秀丽隐杆线虫能够在宽范围的氧浓度下存活，并且生物体的发育进程取决于氧浓度。例如，C.暴露于低氧环境（0.5%至1.5%氧气）的线虫发育缓慢，但暴露于缺氧环境（0%氧气）的线虫在其发育和细胞周期进程中停滞。这种停滞，缺氧诱导的假死，在再次暴露于氧气时是可逆的。除了C以外的生物。斑马鱼、盐水虾和果蝇等秀丽线虫也能够通过阻止发育而在缺氧中存活。 对缺氧引起的发育和细胞周期阻滞知之甚少。为了全面了解缺氧诱导的假死，有必要了解处于不同发育阶段的多种后生动物对缺氧的遗传、生理和细胞反应。暴露于缺氧的胚胎会阻止发育和细胞周期进程。目前尚不清楚缺氧诱导细胞周期停滞是否存在遗传和细胞基础。利用C. elegans研究缺氧诱导的细胞周期停滞将有助于确定是否存在对缺氧的遗传反应。 C. elegans是一个模型系统，其中可以进行经典的正向和反向遗传分析。线虫胚胎的大尺寸和透明性使其成为观察早期发育过程中细胞周期事件的良好系统。 此外，还发现C.线虫的染色体是全着丝粒的，具有延长染色体长度的动粒。因此，在分裂后期，染色体作为一个完整的单位移动，没有落后的臂。此外，C. elegans动粒（最大4微米）使得研究细胞周期事件变得容易。因此，Padilla博士最近发现，线虫胚胎在缺氧条件下会阻止细胞周期进程，这使得线虫成为了解缺氧生存所需的细胞和遗传反应的有用系统。对严重缺氧的反应。这个为期一年的项目的假设是，一个基因（ODS-1），通过RNA干扰（RNAi）筛选的基因是必不可少的C。线虫胚胎在缺氧条件下存活，是线虫胚胎纺锤体检查点的组成部分。 该研究将侧重于ODS-1的表征，并包括以下两个目标：目标1。目的1.确定ODS-1蛋白在常氧和缺氧环境下胚胎中的亚细胞定位。评估ODS-1（RNAi）胚胎的表型。这项研究将导致一个基因和基因产物的表征，该基因和基因产物似乎对线虫胚胎在缺氧中存活很重要，并将允许测试纺锤体检查点参与缺氧诱导的细胞周期停滞的假设。更广泛的影响：这是一个研究启动补助金，授予NSF博士后研究员谁已经接受了终身职位的轨道位置在一个合格的机构，如NSF 00 - 139中所述。 帕迪利亚博士积极整合研究和教育，招募本科生和研究生在她的实验室研究项目的工作，以及在本科课堂教学。