Study of the mitochondrial-cellular response to environmental stress by flourescence imaging

通过荧光成像研究线粒体细胞对环境应激的反应

基本信息

批准号：
9513726
负责人：
GYORGY CSORDAS
金额：
$ 53.56万
依托单位：
THOMAS JEFFERSON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-07-14 至 2020-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9513726
关键词：
Affect Animals Apoptosis Area Arsenic Bioenergetics Cadmium Calcium Calcium Signaling Cardiac Myocytes Cell Culture Techniques Cell Line Cell Respiration Cell Survival Cell physiology Cells Cellular Stress Communication Communities Coupling Cues Dependovirus Development Dioxins Disease Electron Microscopy Endoplasmic Reticulum Generations Goals Heavy Metals HepG2 Hepatic Hepatocyte Homeostasis Image Impairment In Situ Individual Injury Label Light Link Liver Measurement Measures Mediating Methodology Methods Mitochondria Molecular Biology Monitor Morphology Mus Muscle Muscle Fibers Myocardium Organ Organelles Pathogenesis Pathway interactions Phase Population Predisposition Production Proteins Protocols documentation Reactive Oxygen Species Reagent Reporter Research Research Personnel Role Sarcoplasmic Reticulum Scientist Signal Transduction Skeletal Muscle Stress Structure Testing Time Tissues Transgenic Animals Transgenic Mice Viral Work base biological adaptation to stress cell injury environmental agent fluorescence imaging genetic approach in vivo innovation mitochondrial dysfunction mouse model novel oxidative damage public health relevance response sensor technology development tool transgene expression treatment strategy

项目摘要

DESCRIPTION (provided by applicant): A range of environmental agents causes tissue injury that has been attributed to reactive oxygen species (ROS) produced by mitochondria. However, the causative pathways remain largely unknown because it has been difficult to directly monitor or specifically perturb ROS. In the R21 phase, this proposal brings together efforts to develop a new, genetically-targeted toolkit to perturb and measure ROS and calcium (Ca2+) signals in a sensitive and specific manner. Furthermore, this toolkit will allow recording of ROS and Ca2+ down to the level of specific subcompartments of the mitochondria, which likely make differential contributions in ROS dysregulation. The R33 phase will use adeno-associated viruses and transgene expression to bring the novel toolkit into mice to enable study of the effect of various environmental agents on ROS and Ca2+ signals in situ in the liver, heart, and skeletal muscle. Within phases one and two, the project will study the specific involvement of ROS and Ca2+ in the stress pathways triggered by arsenic (As), cadmium (Cd), and dioxin. The investigators will specifically test the novel hypothesis that environmental stress induced by these agents causes impaired mitochondria- endoplasmic/sarcoplasmic reticulum (ER/SR) functional and structural coupling, providing an important mechanism underlying cell injury in various tissues, including the liver, cardiac and skeletal muscle. This team has developed methods to manipulate the mitochondrial-ER/SR interface and has shown that this has consequences for localized Ca2+ signaling. These studies will allow for a paradigm shift in the way mitochondrial pathogenesis of environmental stress is studied, and will shed new light on the mechanisms by which environmental agents can cause tissue damage, leading to unique opportunities for the development of innovative treatment strategies.

描述（由适用提供）：一系列环境药物会导致组织损伤，这归因于线粒体产生的活性氧（ROS）。然而，由于很难直接监测或特别扰动ROS，因此建造的途径仍然在很大程度上未知。在R21阶段，该提案汇集了开发一种新的，遗传靶向的工具包，以敏感和特定的方式扰动和测量ROS和钙（Ca2+）信号。此外，该工具包将允许ROS和Ca2+向下记录到线粒体的特定子组门的水平，这可能会在ROS功能障碍中产生差异贡献。 R33阶段将使用腺相关的病毒和转化表达来将新型工具包带入小鼠中，以研究各种环境药物对ROS和Ca2+信号在第一和第二阶段的原位影响的影响，该项目将研究ROS和Ca2+在Arsenic（ab）中触发的ROS和Ca2+的特定参与。研究人员将特别检验新的假设，即这些药物诱导的环境应激会导致线粒体内质/肌质网（ER/SR）功能和结构耦合受损，从而提供了各种计时损伤的重要机制，包括各种时间，包括脂肪，心脏，心脏，心脏，心脏和骨骼肌肉。该团队已经开发了操纵线粒体/SR接口的方法，并表明这对局部CA2+信号传导有影响。这些研究将允许环境压力的线粒体发病机理的方式发生范式转移，并将对环境药物可能造成组织损害的机制进行新的启示，从而为开发创新治疗策略带来独特的机会。