Genetics/Cell Biology of Anoxia in C.elegans

描述（由申请人提供）：极度缺氧是涉及心脏或肺功能障碍的几种疾病的病理学核心。氧剥夺也在实体肿瘤对放射或化学治疗的抵抗中起作用。了解缺氧抗性生物体对缺氧或低氧的遗传和细胞反应将有助于开发用于拯救受损缺血组织或破坏缺氧肿瘤细胞的治疗。长期的研究目标是确定和表征秀丽隐杆线虫用于缺氧生存的分子机制。 本申请的目的是表征线虫对缺氧的细胞反应，并鉴定缺氧存活所需的基因。本申请的中心假设是线虫具有发育依赖的遗传和细胞机制以在缺氧条件下存活。也就是说，允许胚胎在缺氧中存活的遗传机制对于线虫成虫在缺氧中存活可能并不重要。我们将结合遗传学、细胞生物学和形态学研究来研究C.优雅追求以下具体目标将检验本申请的假设： 目标1.确定ODS-1在缺氧胚胎中的作用ODS-1（Oxygen Deficiency Sensitive）是通过RNA干扰（RNAi）筛选C.线虫胚胎在缺氧条件下存活。ODS-1的蛋白定位和表型分析将被检查，以了解该蛋白在缺氧中的作用。 目标2.识别胚胎在缺氧条件下生存所需的基因。系统的RNAi筛选将用于鉴定胚胎在缺氧或缺氧中存活所需的基因。将检查在该筛选中鉴定的基因的表型。 目标3：描述胚胎后线虫对缺氧的细胞反应，并鉴定胚胎后线虫在缺氧条件下生存所需的基因。为了表征胚胎后野生型线虫对缺氧的反应，我们将使用Nomarski微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筛选来鉴定胚胎后线虫在缺氧或低氧下存活所需的基因。

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.