Detection of Intracellular Oxygen Species

细胞内氧的检测

基本信息

批准号：
8127804
负责人：
SAMEH S ALI
金额：
$ 14.34万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-09-15 至 2012-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8127804
关键词：
Acids Aconitate Hydratase Aconitic Acid Address Aging Antioxidants Apoptosis Apoptotic Behavior Binding Biological Assay Biological Process Biosensing Techniques Biosensor Brain Calibration Cell Culture Techniques Cell Extracts Cells Cerebral Hypoxia Cerebral Ischemia Complement Craniocerebral Trauma Cytosol Data Detection Development Disease Dose Electrodes Electron Spin Resonance Spectroscopy Electron Transport Electrons Employment Environment Enzyme Electrodes Enzymes Etiology Exposure to Fluorescence Microscopy Free Radicals Functional disorder Genetic Transcription Glucose Goals Heart Arrest Homeostasis Hydrogen Peroxide Hydroxyl Radical Hypoxia Immobilized Enzymes Immune response Individual Injury Intervention Iron Isocitrates Kinetics Knockout Mice Life Lipids Location Measures Membrane Membrane Fluidity Membrane Potentials Methodology Methods Mitochondria Mitochondrial Matrix Monitor N-Methylaspartate Neuronal Injury Neurons Oligodendroglia Optics Organism Outcome Oxidation-Reduction Oxygen Oxygen saturation measurement Pathologic Physical environment Physiological Play Principal Investigator Production Reactive Oxygen Species Research Research Personnel Role SOD2 gene Serum Signal Transduction Site Sodium Chloride Specific qualifier value Spin Trapping Stimulus Stroke Structure of superior cervical ganglion Sulfhydryl Compounds Superoxide Dismutase Superoxides System Techniques Therapeutic Training Transduction Gene Vitamin E Withdrawal adduct age related arctic environment base biological systems career cold temperature cytochrome c deprivation detector dihydroethidium extracellular fluorescence imaging healthy aging improved isocitrate lipophilicity nanobiosensor nanofiber nanosensors neurotoxic oxidation programs research study sensor simulation tetracyanoquinodimethane tetrathiafulvalene

项目摘要

DESCRIPTION (provided by applicant): Detection of Intracellular Oxygen Species Among the fundamental questions in research on the biomolecular basis of disease is whether reactive oxygen species (ROS), e.g. superoxide (O2-) and H2O2, are continuously produced by cells -from mitochondria or other sites, and what levels of these ROS are generated under physiological and pathological conditions. An essential requirement to address this is development of noninvasive techniques capable of assaying O2- and H2O2 at the cellular and subcellular level. The long-term objective of the present research is to develop quantitiative techniques to detect and quantify ROS in single cells, with a focus on mitochondriaL ROS production, and to correlate results from these techniques with commonly-used assays for ROS such as fluorescence microscopy using oxidation-sensitive probes. To achieve these goals, we will exploit developments in the fields of electrochemical nanosensors and spin-trapping electron paramagnetic resonance (EPR) spectroscopy. Studies will be carried out in cell cultures and mitochondria using the following approach: 1) synthesize and apply new lipophilic derivatives of the spin trap DEPMPO to trap O2-, 2) correlate EPR spectra and data from EPR oximetry to obtain information about the environment in which O2- is released, 3) characterize nanosensors based on covalently attached enzymes on functionalized optical nanofibers for intracellular detection of O2- and/or H2O2 , 4) compare results from these techniques with those from more accessible methods, e.g. fluorescence microscopy, and 5) study changes in O2- and H2O2 levels under injury conditions. It is hoped that these studies will address questions such as: i) What are actual concentrations of O2- and H2O2 after physiologic and pathologic stimuli? ii) Are specific diseases associated with elevated vs. decreased ROS levels? and iii) What are the best therapeutic strategies to ameliorate or reverse changes in ROS without compromising their signaling functions?

描述（由申请人提供）：细胞内氧的检测疾病的生物分子基础研究中的基本问题之一是活性氧（ROS）是否存在于细胞内。超氧化物 (O2-) 和 H2O2 由细胞从线粒体或其他部位持续产生，以及在生理和病理条件下产生的这些 ROS 水平是多少。解决这个问题的一个基本要求是开发能够在细胞和亚细胞水平上测定 O2- 和 H2O2 的非侵入性技术。本研究的长期目标是开发定量技术来检测和量化单细胞中的 ROS，重点关注线粒体 ROS 的产生，并将这些技术的结果与常用的 ROS 测定（例如使用荧光显微镜）关联起来。氧化敏感探针。为了实现这些目标，我们将利用电化学纳米传感器和自旋捕获电子顺磁共振（EPR）光谱学领域的发展。将使用以下方法在细胞培养物和线粒体中进行研究：1) 合成并应用自旋捕获 DEPMPO 的新亲脂性衍生物来捕获 O2-，2) 将 EPR 光谱和 EPR 血氧测定法的数据关联起来，以获得有关环境的信息。释放 O2-，3) 表征基于功能化光学纳米纤维上共价连接酶的纳米传感器，用于细胞内检测 O2- 和/或 H2O2，4)将这些技术的结果与更容易使用的方法（例如，荧光显微镜，5) 研究损伤条件下 O2- 和 H2O2 水平的变化。希望这些研究能够解决以下问题： i) 生理和病理刺激后 O2- 和 H2O2 的实际浓度是多少？ ii) 特定疾病是否与 ROS 水平升高或降低相关？ iii) 在不损害其信号功能的情况下改善或逆转 ROS 变化的最佳治疗策略是什么？

项目成果

期刊论文数量（1）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Loss of caveolin-1 accelerates neurodegeneration and aging.

Caveolin-1 的缺失会加速神经退行性变和衰老。

DOI：
发表时间：
2010-12-23
期刊：
影响因子：
3.7
作者：
Head, Brian P;Peart, Jason N;Panneerselvam, Mathivadhani;Yokoyama, Takaakira;Pearn, Matthew L;Niesman, Ingrid R;Bonds, Jacqueline A;Schilling, Jan M;Miyanohara, Atsushi;Headrick, John;Ali, Sameh S;Roth, David M;Patel, Piyush M;Patel, Hemal H
通讯作者：
Patel, Hemal H