Neuropeptides in the CNS with New Mass Spectrometric Sampling Protocols

中枢神经系统中的神经肽采用新的质谱采样方案

• 批准号: 7618153
• 负责人: Jonathan V. Sweedler
• 金额: $ 21.17万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-08-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7618153
• 关键词: Address; Architecture; Area; Award; Basic Science; Biochemical; Biochemistry; Biological; Brain; Brain region; Cell Culture Techniques; Cell Separation; Cells; Cerebellar cortex structure; Chemical Stimulation; Chemical Structure; Chemicals; Collection; Complex; Corpus Callosum; Detection; Development; Drug Addiction; Drug abuse; Exposure to; Follow-Up Studies; Goals; Individual; Investigation; Learning; Location; Mammalian Cell; Maps; Mass Spectrum Analysis; Measurement; Measures; Methods; Nerve Tissue; Nervous system structure; Neurobiology; Neuromodulator; Neurons; Neuropeptides; Neurosciences; Outcome; Output; Peptide Signal Sequences; Peptide Synthesis; Peptide Transport; Peptides; Phase; Physiological; Post-Translational Protein Processing; Preparation; Principal Investigator; Process; Proteins; Proteomics; Protocols documentation; Rattus; Research; Research Design; Research Personnel; Sampling; Signaling Molecule; Slice; Solid; Spatial Distribution; Specificity; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Staging; Techniques; Technology; Testing; Time; Tissues; Validation; amidation; base; biological systems; cell growth; cytokine; drug of abuse; genetic analysis; insight; interest; member; neuronal cell body; new technology; novel; programs; relating to nervous system; research study; technique development; technology development; tool; Address; Architecture; Area; Award; Basic Science; Biochemical; Biochemistry; Biological; Brain; Brain region; Cell Culture Techniques; Cell Separation; Cells; Cerebellar cortex structure; Chemical Stimulation; Chemical Structure; Chemicals; Collection; Complex; Corpus Callosum; Detection; Development; Drug Addiction; Drug abuse; Exposure to; Follow-Up Studies; Goals; Individual; Investigation; Learning; Location; Mammalian Cell; Maps; Mass Spectrum Analysis; Measurement; Measures; Methods; Nerve Tissue; Nervous system structure; Neurobiology; Neuromodulator; Neurons; Neuropeptides; Neurosciences; Outcome; Output; Peptide Signal Sequences; Peptide Synthesis; Peptide Transport; Peptides; Phase; Physiological; Post-Translational Protein Processing; Preparation; Principal Investigator; Process; Proteins; Proteomics; Protocols documentation; Rattus; Research; Research Design; Research Personnel; Sampling; Signaling Molecule; Slice; Solid; Spatial Distribution; Specificity; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Staging; Techniques; Technology; Testing; Time; Tissues; Validation; amidation; base; biological systems; cell growth; cytokine; drug of abuse; genetic analysis; insight; interest; member; neuronal cell body; new technology; novel; programs; relating to nervous system; research study; technique development; technology development; tool

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.

描述（由申请人提供）：药物滥用研究的一个重要目标是在暴露于滥用药物的情况下检测和表征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较高神经元功能联系起来。在众多信号分子中，细胞间信号肽（ISP）是一个巨大的测量挑战。在生物系统中，实际上有数百种肽，其中许多肽仅在翻译后修饰时具有生理活性。拟议的研究都开发并实施了基于基质的激光解吸/电离质谱法（MALDI-MS）的技术来发现和表征ISP的技术，包括确定其化学形式和哺乳动物神经系统中的空间定位。总体策略是开发新技术和采样方案，以检测和鉴定非常小的生物样品中的新ISP，包括新鲜制备或培养的单个哺乳动物细胞，小组细胞以及从神经系统的空间限制区域释放的ISP。这些技术承诺将揭示数百种潜在的ISP，这些ISP将通过旨在：（a）找到具有特定后翻译后修饰的肽（例如胺化）； （b）确定神经系统中经历长距离传输的肽； （c）发现哪些肽是以活动依赖性方式释放的。这项技术开发理想地与尖端的基础研究奖邀请相匹配。通过开发强大的基于蛋白质组学的技术来回答有关最复杂，最难以捉摸的神经调节剂的问题，神经肽，对滥用药物对中枢神经系统的影响的基本见解将获得。