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.

氧气剥夺会影响生物的生长和发育。自然界中的许多生物都会发生氧气水平的变化，并适应氧气剥夺。土壤线虫秀丽隐杆线虫能够存活各种氧气水平，生物体的发育进展取决于氧气浓度。例如，暴露于低氧环境（0.5％至1.5％的氧气）的秀丽隐杆线虫会缓慢发展，但在其发育和细胞周期进程中暴露于缺氧环境（0％氧）停滞的线虫。这种逮捕是缺氧引起的悬浮动画，在重新暴露于氧气后是可逆的。秀丽隐杆线虫以外的生物，例如斑马鱼，盐水虾和水果蝇，也能够通过阻止发育来生存缺氧。 关于响应缺氧的发育和细胞周期停滞知之甚少。为了充分了解缺氧引起的悬浮动画，有必要了解在发育的不同阶段，在各种后生动物中对缺氧的遗传，生理和细胞反应。尚不清楚是否存在缺氧诱导细胞周期停滞的遗传和细胞基础。使用诸如秀丽隐杆线虫等遗传模型系统研究缺氧引起的细胞周期停滞将有助于确定是否对缺氧有遗传反应。 秀丽隐杆线虫是一个模型系统，在该系统中，经典的前进和反向遗传分析是可能的。线虫胚胎的较大尺寸和透明度使它们成为观察早期发育过程中细胞周期事件的绝佳系统。 此外，秀丽隐杆线虫染色体是全中心的，具有延伸染色体长度的动力学。因此，在后期期间，染色体随着整个单元而无需滞留的臂移动。此外，大尺寸的秀丽隐杆线虫动物（最多4微米）使研究细胞周期事件变得容易。因此，帕迪拉博士最近发现线虫胚胎会阻止缺氧的细胞周期进展，使线虫成为理解氧剥夺生存所需的细胞和遗传反应的有用系统。帕迪拉（Padilla）的长期研究目标是确定秀丽隐杆线虫对严重氧气剥夺的反应的机制。这个为期一年的项目的假设是，通过RNA干扰（RNAi）筛选的基因（ODS-1）对于秀丽隐杆线虫胚胎生存的缺氧至关重要的基因是线虫胚胎中纺锤体检查点的一个组成部分。 这项研究将侧重于ODS-1的表征，并由以下两个目的组成：目标1。确定在暴露于常氧或缺氧环境的胚胎中ODS-1蛋白的亚细胞定位。IAM2。评估ODS-1（RNAI）胚胎的表型的表型将导致胚胎的胚胎对基因和基因产品的表征，以使其对基因的表征进行表征，以使其在基因上的表征，以使其在基因上的表征，以使其在基因上的表征有效，以使其在基因上的表征，以使其出现，以表征出现的基因，以使其在基因上的表征，以使其出现的eNS emem n n n e Engiiip.并将允许检验假设，即主轴检查点与缺氧引起的细胞周期逮捕有关。Boader的影响：这是一项研究起动器赠款，授予NSF博士后研究员，他在NSF 00-139中所述，在合格机构中接受了合格机构的任期职位。 帕迪拉（Padilla）博士通过招募本科生和研究生来积极整合研究和教育，以在她的实验室以及在本科教室中的教学中从事研究项目。