Genetics/Cell Biology of Anoxia in C.elegans

描述(由申请人提供)：极端缺氧是几种涉及心脏或肺功能障碍的疾病的病理中心。缺氧在实体肿瘤对放射或化疗的抵抗中也起到了作用。了解耐缺氧生物对缺氧或缺氧的遗传和细胞反应，将有助于开发抢救受损的缺血组织或破坏缺氧的肿瘤细胞的治疗方法。长期的研究目标是识别和表征秀丽线虫在缺氧条件下生存的分子机制。 这项应用的目的是描述线虫对缺氧的细胞反应，并识别缺氧生存所需的基因。这一应用的中心假设是，线虫具有发育依赖的遗传和细胞机制，可以在缺氧条件下生存。也就是说，允许胚胎在缺氧中存活的遗传机制可能对线虫成虫在缺氧中存活并不重要。我们将结合遗传学、细胞生物学和形态学研究来研究线虫的缺氧。追求以下具体目标将检验这项申请的假设： 目的1.确定ODS-1在缺氧胚胎中的作用。通过RNA干扰(RNAi)筛选线虫胚胎在缺氧条件下存活所必需的基因，鉴定出了对氧剥夺敏感的ODS-1。我们将研究消耗臭氧层物质-1的蛋白质定位和表型分析，以了解该蛋白质在缺氧中所起的作用。 目的2.确定胚胎缺氧生存所需的基因。系统的RNAi筛查将用于识别胚胎在缺氧或缺氧状态下存活所需的基因。将检查在此筛查中确定的基因的表型。 目的3.研究胚胎后线虫对缺氧的细胞反应，并鉴定胚胎后线虫缺氧生存所需的基因。为了表征胚胎后野生型线虫对缺氧的反应，我们将使用诺马尔斯基差分干涉对比光学显微镜和特定组织中绿色荧光蛋白的可视化来分析组织。我们将使用系统的RNAi筛选来识别胚胎后线虫缺氧或缺氧生存所需的基因。

DESCRIPTION (provided by applicant): Extreme oxygen deprivation is central to the pathology of several diseases involving cardiac or pulmonary dysfunction. Oxygen deprivation also plays a role in the resistance of solid tumors to radiation or chemotherapy treatment. Understanding the genetic and cellular response oxygen-deprivation resistant organisms have to anoxia or hypoxia will facilitate the development of treatment for the rescue of damaged ischemic tissue or the destruction of oxygen deprived tumor cells. The long-term research goal is to identify and characterize the molecular mechanisms Caenorhabditis elegans use to survive oxygen deprivation. The objectives of this application are to characterize the cellular response nematodes have to oxygen deprivation and to identify genes required for oxygen deprivation survival. The central hypothesis of this application is that nematodes have developmentally dependent genetic and cellular mechanisms to survive oxygen deprivation. That is, a genetic mechanism that allows embryos to survive anoxia may not be important for the nematode adult to survive anoxia. We will use a combination of genetic, cell biological, and morphological studies to study oxygen deprivation in C. elegans. Pursuing the following specific aims will test the hypothesis of this application: Aim 1. Determine the role ODS-1 has in anoxic embryos. ODS-1 (Oxygen Deprivation Sensitive) was identified by a RNA interference (RNAi) screen for genes that are essential for C. elegans embryos to survive anoxia. The protein localization and phenotypic analysis of ODS-1 will be examined to understand the role this protein has in anoxia. Aim 2. Identify genes required for embryos to survive oxygen deprivation. A systematic RNAi screen will be used to identify genes required for embryos to survive anoxia or hypoxia. The phenotype of genes identified in this screen will be examined. Aim 3. Characterize the cellular response post-embryonic nematodes have to oxygen deprivation and identify genes required for post-embryonic nematodes to survive oxygen deprivation. To characterize the response post-embryonic wild type nematodes have to oxygen deprivation we will analyze tissues using Nomarski differential-interference contrast optical microscopy and visualization of green fluorescent protein in specific tissues. We will use systematic RNAi screen to identify genes required for post-embryonic nematodes to survive anoxia or hypoxia.