NMR STUDIES OF CELLULAR METABOLISM

细胞代谢的核磁共振研究

• 批准号: 3855925
• 负责人: R LONDON
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ENVIRONMENTAL HEALTH SCIENCES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3855925
• 关键词: boric acids; boron; dogs; erythrocytes; fluorescent dye /probe; fluorine; halocarbon compound; laboratory rat; nuclear magnetic resonance spectroscopy; oxygen tension; reagent /indicator; toxin metabolism

This program is aimed at the development and application of in vivo NMR spectroscopic methods for studying metabolism and its perturbation by chemical toxins. A principal focus of these studies has been the development of NMR active, intracellular indicator molecules to allow determination of metabolic parameters of interest in intact cells. Research has focused on the use of fluorinated indicators as a consequence of the inherent sensitivity of fluorine for NMR detection and the essential absence of background fluorine resonances from untreated cells. During the past year, progress was made in the development of fluorinated indicators for cytosolic calcium, magnesium, and pH. One of the fluorinated pH indicators synthesized during the past year appears to have more ideal kinetic, shift sensitivity, and loading properties than previously described pH indicators, and should provide valuable insight into this important metabolic parameter. We have also continued work on the use of perfluorocarbons, several of which are used clinically in surgical eye procedures, as indicators of cellular oxygen concentration, and evaluated the sensitivity of the fluorine chemical shift values to temperature as well. It appears that both oxygen concentration and temperature can be determined from NMR measurements on tissues containing these perfluorocarbons. A new effort initiated over the past year is aimed at determining the molecular basis for the toxicity of borate and several organic boron derivatives. Boric acid, which has a number of important commercial applications, has been shown to cause testicular atrophy in rats and dogs after ingestion of diets containing > 1000 ppm. During the past year we have carried out NMR studies of boric acid and phenyl boric acid in the presence of intact erythrocytes, enzymes, and other cellular materials thought to be potential targets, in order to quantify and evaluate the biochemical significance of these interactions. In addition, studies of the carbohydrate metabolism of sertoli cells using [1-13C] and [2-13C] labeled glucose have been initiated to investigated possible alterations related to borate toxicity.

本项目是针对活体核磁共振的发展和应用而开发的 新陈代谢及其扰动的光谱研究方法 化学毒素。这些研究的一个主要焦点是 核磁共振活性细胞内指示剂分子的开发 完整细胞中感兴趣的代谢参数的测定。 因此，研究的重点是氟化指示剂的使用 核磁共振检测中氟的固有灵敏度及其本质 未经处理的细胞没有本底氟共振。在.期间 过去一年，在开发氟化指标方面取得了进展 胞浆中的钙、镁和pH值。一种氟化的pH值 过去一年综合的指标似乎更理想 与以前相比，动力学、变速灵敏度和加载特性 描述了酸碱度指标，并应对此提供有价值的见解 重要的代谢参数。我们还继续就使用 全氟碳化合物，其中几种临床用于外科手术眼 程序，作为细胞氧浓度的指示器，并评估 氟化学位移值对温度的敏感性 井。看起来氧气浓度和温度都可以 根据对含有这些物质的组织的核磁共振测量确定的 全氟碳化合物。过去一年发起的一项新努力旨在 确定硼酸盐和几种化合物毒性的分子基础 有机硼衍生物。硼酸，它有许多重要的 商业应用，已被证明可导致大鼠睾丸萎缩 和狗在摄入含有1000ppm的饲料后。在过去 一年来，我们对硼酸和苯基硼酸进行了核磁共振研究。 完整的红细胞、酶和其他细胞物质的存在 被认为是潜在的目标，以便量化和评估 这些相互作用的生物化学意义。此外，还研究了 支持细胞[1-13C]和[2-13C]的碳水化合物代谢 已经开始标记葡萄糖来研究可能的变化 与硼酸盐毒性有关。