Sampling and Detection of Neuropeptides in nL Volumes

描述：（申请人提供） 阿片肽（脑啡肽和强啡肽）在脑内的功能作用 药物滥用和成瘾问题已得到广泛承认，但尚未得到充分理解。 如果能提供关于以下方面的资料，将促进这一领域的进展： 细胞外阿片肽的浓度、寿命和调节 活脑的空间该研究项目的目标是克服 这些技术挑战现在阻碍了获取这些信息。 必须克服两个技术障碍。首先，阿片肽存在于低水平 细胞外空间的水平。其次，它们作为肽家族存在 而不是单独的、不同的物质。因此，分析方法与 特别高的灵敏度和优异的选择性是必需的，但 尚未广泛使用。在此期间开发的敏感和选择性方法 这个项目将建立在一个最先进的平台上， 可用的色谱和质谱仪器。所以他们 将很容易被众多研究实验室采用， 药物滥用的石化方面。 将使用毛细管移液器收集少量未稀释的纯 大鼠纹状体组织切片的细胞外液。肽含量 将在基础条件下采集的样本中测定， 钾诱导的去极化，在不存在钙离子的情况下， 存在肽酶抑制剂。

DESCRIPTION: (provided by the applicant) The functional role of the opioid peptides (encephalin and dynorphins) in substance abuse and addiction is well recognized but not fully understood. Advances in this area would be stimulated by information regarding the concentration, lifetime, and regulation of opioid peptides in the extracellular space of the living brain. The goal of this research project is to overcome the technical challenges that now obstruct access to that information. Two technical obstacles must be overcome. First, opioid peptides exist at low levels in the extracellular space. Second, they exist as families of peptides rather than individual, distinct substances. Thus, analytical methods with exceptionally high sensitivity and excellent selectivity are necessary but are not yet widely available. The sensitive and selective methods developed during this project will be built on a platform of state-of-the-art, commercially available chromatographic and mass spectrometric instrumentation. Thus, they will be readily adoptable by the numerous research laboratories addressing the petrochemical aspects of substance abuse. A capillary pipet will be used to collect small, undiluted samples of neat extracellular fluid of striatal tissue slices from rats. The peptide content will be determined in samples acquired under basal conditions, during potassium-induced depolarization, in the absence of calcium ion, and in the presence of peptidase inhibitors.