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.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 科夫曼实验室利用其INBRE支持（启动资金授予詹姆斯科夫曼，PI） 开展口腔发育特化的机制研究， 海胆幼虫的反口轴。 轴规范是初始对称破缺 为有机体的身体平面建立空间坐标的发育过程。 基于以前的工作，我们假设海胆幼虫的OA轴是 最初由氧化还原梯度确定，该氧化还原梯度是由线粒体的不对称分布确定的， 卵和早期胚胎，胚胎的一侧遗传了最高密度的 线粒体倾向于激活TGF-β信号分子的表达 结状的，并由此发育为口腔外胚层。 我们进一步假设， OA极性涉及线粒体H2 O2介导的磷酸酶失活， 否则使Nodal表达所需的p38-MAPK失活。 期间 在这一资助期间，我们开发了测试这些假设的工具，并利用它们来 为未决的NIH/NIEHS拨款申请生成初步数据。 这些工具包括 靶向氧化还原调节蛋白磷酸酶的吗啉代反义寡核苷酸， PP 2A，我们的初步数据表明其位于p38-MAPK和Nodal表达的上游; 我们用它来追踪线粒体在细胞中的分布， 活的胚胎;以及一种以肾脏为靶点的过氧化氢酶，一种消耗H2 O2的酶 我们用它来检验线粒体H2 O2是一种信号传导的假设， p38活化和随后的口腔外胚层特化所需的中间体。 这些试剂也将用于检验假设，即众所周知的OA敏感性 对环境毒物（如重金属）的轴质量标准可归因于 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拨款申请。