NEUROPEPTIDES IN THE CNS WITH IMAGING MASS SPECTROMETRY

描述（由申请人提供）：药物滥用研究的一个重要目标是检测和表征暴露于滥用药物后神经系统生物化学的变化，并将这些变化与正常和病理条件下的高级神经元功能联系起来。在众多的信号分子中，细胞间信号肽（ISP）是一个巨大的测量挑战;在生物系统中表达的肽有数百种，其中许多只有在后期修饰时才具有生理活性。拟议的研究开发和实施基于基质辅助激光解吸/电离质谱（MALDI-MS）的技术，用于发现和表征ISPs，包括确定其化学形式和哺乳动物神经系统中的空间定位。总体战略是开发新技术和采样协议，以检测和识别非常小的生物样品中的新ISP，包括单独的新鲜制备或培养的单个哺乳动物细胞，小细胞群，以及从神经系统空间受限区域释放的细胞。这些技术有望发现数百种潜在的ISP，这些ISP将通过设计用于以下目的的研究来过滤功能相关性：（A）发现具有特定翻译后修饰的肽（例如，酰胺化）;（B）确定在神经系统中经历长距离运输的肽;（C）发现哪些肽以活性依赖性方式释放。这一技术发展与尖端基础研究奖的征集工作完美匹配。通过开发强大的新的基于蛋白质组学的技术来回答关于最复杂和最难以捉摸的神经调质，神经肽的问题，将获得对滥用药物对CNS影响的基本见解。

DESCRIPTION (provided by applicant): An important goal of drug abuse research is to detect and characterize changes in the biochemistry of the nervous system on upon exposure to drugs of abuse, and to relate these changes to higher neuronal functions in normal and pathological conditions. Amongst numerous signaling molecules, intercellular signaling peptides (ISPs) are a dramatic measurement challenge; there are literally hundreds of peptides expressed in biological systems, many of which are only physiologically active when posttranslationally modified. The proposed research both develops and implements matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) based technologies for the discovery and characterization of ISPs, including determination of their chemical form and spatial localization in the mammalian nervous system. The overall strategy is to develop new technologies and sampling protocols to detect and identify new ISPs in very small biological samples, including individual freshly prepared or cultured single mammalian cells, small groups of cells, and those released from spatially restricted regions of the nervous system. These technologies promise to uncover hundreds of potential ISPs that will be filtered for functional relevance by studies designed to: (A) find peptides with specific post- translational modifications (e.g., amidation); (B) determine the peptides that undergo long- distance transport in the nervous system; (C) discover which peptides are released in an activity-dependent manner. This technology development ideally matches the cutting-edge basic research award solicitation. By developing powerful new proteomics-based technologies to answer questions about the most complex and elusive set of neuromodulators, the neuropeptides, basic insight into the effects of drugs of abuse on the CNS will be gained.