MEASURMENT OF DAG MOLECULAR SPECIES IN BASOPHILS

描述(改编自调查者摘要)：二酰基甘油 (DAG)在早期受体介导的信号转导中发挥核心作用 许多类型的单元格的事件。这些脂质分子似乎也是 参与细胞分化、细胞生长和细胞凋亡。先前 现有的对这类分子的分析只在 微克分子范围，如果检查了总DAG水平，或需要显著 更高的水平，如果一个人对分子物种概况感兴趣的话。那里 有许多关于DAG物种的相对作用以及 关于它们产生的时间和地点的问题。此应用程序寻求 为了改进一种测量二甘油生成的新技术， 酰基或烷基连接的。这项技术的灵敏度提高了1000倍 与常规方法相比，分子物种特异性和质量 对这些物种的测量。该方法使用气体 色谱/负离子化学电离质谱仪 (GC/NICIMS)分离和测定选定的DAG分子物种 用于定量的内部氢化标准。该应用程序寻求 充分发展这一方法，使DAG和DAG的分子物种 磷脂可以高灵敏度地分析。此外，一个新的 方法来描述磷脂物种，而不需要事先知道 提出了物种存在的观点。当后一种方法完全发展时，它 或许有可能预测哪些DAG物种值得关注 其他建议的方法。后一种技术应该允许 研究人员了解细胞选择性产生的程度 特定的分子物种。我们自然期待有一个高度的 对DAG的敏感检测将促进对这一重要第二阶段的研究 信使存在于各种各样的细胞系统和细胞器中。

DESCRIPTION (Adapted from the Investigator's abstract): Diacylglycerols (DAG) have a central role in early receptor-mediated signal transduction events for many types of cells. These lipid molecules also appear to be involved in cell differentiation, cell growth and apoptosis. Previously existing assays for this class of molecules are sensitive only in the picomole range, if total DAG levels are examined, or require significantly higher levels if one is interested in a molecular species profile. There are many questions regarding the relative role of DAG species as well as questions about when and where they are generated. This application seeks to refine a new technique to measure the generation of diradylglycerols, acyl or alkyl linked. This technique provides 1000 fold better sensitivity than conventional methods, molecular species specificity and mass measurements of these species. The method employs gas chromatography/negative ion chemical ionization mass spectrometry (GC/NICIMS) to separate and measure selected molecular species of DAG using internal deuterated standards for quantitation. The application seeks to fully develop this approach so that both molecular species of DAG and phospholipids may be analyzed with high sensitivity. In addition, a new method to profile phospholipid species, without a prior knowledge of the species present is proposed. When this latter method is fully developed, it may be possible to predict which DAG species deserve attention using our other proposed methodologies. This latter technique should allow investigators to understand the extent to which cells selectively generate specific molecular species. We naturally expect that having a highly sensitive assay for DAGs will facilitate studies of this important second messenger in a wide variety of cells systems and cell organelles.