ORAL-ABORAL AXIS SPECIFICATION IN THE SEA URCHIN EMBRYO

海胆胚胎的口腔轴规格

• 批准号: 7610081
• 负责人: JAMES A COFFMAN
• 金额: $ 4.54万
• 依托单位: MOUNT DESERT ISLAND BIOLOGICAL LAB
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-01 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7610081
• 关键词: Antisense Oligonucleotides; Applications Grants; Award; Computer Retrieval of Information on Scientific Projects Database; Data; Development; Embryo; Enzymes; Event; Funding; Grant; Green Fluorescent Proteins; Heavy Metals; Hydrogen Peroxide; Inherited; Institution; Larva; Life; MAPK14 gene; Mediating; Mitochondria; Nodal; Oral; Organism; Oxidants; Oxidation-Reduction; Phosphoric Monoester Hydrolases; Production; Protein Phosphatase 2A Regulatory Subunit PR53; Protein phosphatase; Reagent; Research; Research Personnel; Resources; Sea Urchins; Side; Signal Transduction; Signaling Molecule; Source; Specific qualifier value; Testing; Toxic Environmental Substances; Transforming Growth Factor beta; United States National Institutes of Health; Work; base; catalase; density; egg; oral ectoderm; tool

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. The Coffman Lab has used its INBRE support (startup funds awarded to James Coffman, PI) to carry out research on the mechanism underlying developmental specification of the oral- aboral (OA) axis of the sea urchin larva. Axis specification is the initial symmetry breaking event in development that sets up the spatial coordinates for the body plan of an organism. Based on previous work, we have hypothesized that the OA axis of the sea urchin larva is initially specified by a redox gradient established by asymmetric distribution of mitochondria in the egg and early embryo, with the side of the embryo inheriting the highest density of mitochondria being pre-disposed to activate expression of the TGF-beta signaling molecule Nodal and thence develop as oral ectoderm. We further hypothesize that the specification of OA polarity involves mitochondrial H2O2-mediated inactivation of a phosphatase that otherwise inactivates p38-MAPK, activity of which is required for Nodal expression. During this funding period we have developed tools to test these hypotheses, and have used them to generate preliminary data for a pending NIH/NIEHS grant application. These tools include morpholino antisense oligonucleotides targeted to a redox-regulated protein phosphatase, PP2A, which our preliminary data suggest lies upstream of p38-MAPK and Nodal expression; a mitochondrially targeted GFP, which we are using to track the distribution of mitochondria in living embryos; and a mitochondrially targeted catalase, an enzyme that depletes H2O2 which we are using to test the hypothesis that mitochondrial H2O2 is a signaling intermediate required for p38 activation and consequent specification of oral ectoderm. These reagents will also be used to test the hypothesis that the well known sensitivity of OA axis specification to environmental toxicants such as heavy metals is attributable to alterations in redox state and mitochondrial oxidant production, which is the focus of our pending NIEHS grant application.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 科夫曼实验室（Coffman Lab）使用了近亲支持（授予詹姆斯·科夫曼（James Coffman）的启动资金） 对口腔发育规范的基础机制进行研究 海胆幼虫的原住民（OA）轴。 轴规范是初始对称性断裂 开发的事件为有机体的身体计划建立了空间坐标。 根据以前的工作，我们假设海胆幼虫的OA轴是 最初是由通过线粒体不对称分布确定的氧化还原梯度指定的 卵和早期胚胎，胚胎的一侧继承了最高密度 线粒体预先介绍以激活TGF-β信号分子的表达 淋巴结和随着口服外胚层的发展。 我们进一步假设 OA极性涉及线粒体H2O2介导的磷酸酶的失活，该磷酸酶的失活 否则会使p38-mapk失活，淋巴结表达需要其活性。 期间 在这个资金期间，我们开发了测试这些假设的工具，并使用它们来 为待处理的NIH/NIEHS赠款应用程序生成初步数据。 这些工具包括 靶向氧化还原调节的蛋白磷酸酶的吗啡反义寡核苷酸， pp2a，我们的初步数据表明，p38-mapk和节点表达的上游；一个 线粒体靶向的GFP，我们用来跟踪线粒体在中的分布 活着的胚胎；线粒体靶向过氧化氢酶，一种耗尽H2O2的酶 我们用来测试线粒体H2O2是一个信号传导的假设 p38激活所需的中间体和口服外胚层的规范。 这些试剂还将用于检验以下假设：OA的敏感性 轴向环境有毒物质（例如重金属）的规范归因于 氧化还原状态和线粒体氧化剂产生的改变，这是我们的重点 等待NIEHS赠款申请。