In Situ Quantitation of Neuropeptides in Feeding

饲喂中神经肽的原位定量

• 批准号: 7219873
• 负责人: Stephanie Shannon Cape
• 金额: $ 2.84万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-01-15 至 2007-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7219873
• 关键词: Affect; Animals; Beds; Biological; Biological Models; Biological Neural Networks; Cardiovascular Diseases; Cells; Class; Complex; Coupled; Crabs; Crustacea; Desire for food; Detection; Development; Eating; Eating Disorders; End Point; Energy Metabolism; Ensure; Feeding behaviors; Food deprivation (experimental); Fourier Transform; Ganglia; Health; Homeostasis; In Situ; Incidence; Investigation; Isotope Labeling; Label; Lead; Liquid substance; Malignant Neoplasms; Marriage; Mass Spectrum Analysis; Measures; Methodology; Methods; Modeling; Molecular; Myxoid cyst; Nature; Nervous system structure; Neurons; Neuropeptides; Neurosecretory Systems; Non-Insulin-Dependent Diabetes Mellitus; Obesity; Organ; Organism; Peptides; Physiological; Physiological Processes; Play; Pliability; Process; Property; Proteins; Proteomics; Regulation; Relative (related person); Reproducibility; Research; Risk; Role; Sampling; Scheme; Signal Pathway; Signal Transduction; Signaling Molecule; Solutions; Stable Isotope Labeling; Stroke; System; Techniques; Testing; Tissues; Validation; analytical method; base; cost; feeding; health economics; improved; innovation; insight; interest; mRNA Differential Displays; mass spectrometer; method development; neurochemistry; novel; technology development; tissue processing

DESCRIPTION (provided by applicant): the health and economic costs of eating disorders are enormous. For example, obesity is associated with type 2 diabetes, cardiovascular disease, stroke, and cancer. Increasingly, the importance of the role of neuropeptides in appetite and energy'expenditure is being established. However, the high complexity of signaling pathways that regulate food intake and energy homeostasis makes the study of these critical mechanisms daunting. Therefore, it is not only necessary to develop advanced analytical methods to measure and identify these low abundance signaling molecules, but also to apply these methods to study the feeding circuit in a simpler well-defined system. The specific aims of this project include: (1) To develop novel mass spectrometry based strategies to enable enhanced in situ neuropeptide profiling and the first demonstration of quantitative direct tissue analysis and (2) To apply these innovative methods to investigate the differential display of neuropeptides in satiated and food deprived Cancer borealis (Jonah crab). The development of a mass spectrometry based method that allows quantitative comparison of neuropeptides in situ will enable increased sampling flexibility and reduced tissue processing thus decreasing the risk of analyte loss or contamination. Not only does the crustacean nervous system offer an excellent test-bed for this technology development and validation, but it is also an ideal model for investigating neuromodulation in a well-defined neural network. A high incidence of identified neurons whose electrophysiological properties can be readily assessed and a rich repertoire of neuromodulatory substances makes this small neural network an attractive model system. This project has great potential to lead to the discovery of new neuropeptides and increase understanding of the neurochemical basis of the peptidergic regulation of feeding behavior. Not only will the analytical methods developed be applicable to the study of larger, more complex vertebrate systems, but also the molecular insights gained from the study of the crustacean system will likely lead to the identification of homologous mechanisms in higher order species. Therefore, this study will lead to improved methods and extended capabilities for neuropeptide analysis as well as the identification of potential targets for the development of new pharmacologic treatments for eating disorders. *New methods for measuring molecular signals used by the nervous system will be developed and then applied to study how feeding behavior is controlled. This research will lead to a better understanding of nervous system principles as well as new treatments for important health concerns such as obesity.

描述（由申请人提供）：饮食失调的健康和经济成本是巨大的。例如，肥胖与 2 型糖尿病、心血管疾病、中风和癌症有关。人们日益认识到神经肽在食欲和能量消耗中的作用的重要性。然而，调节食物摄入和能量稳态的信号通路的高度复杂性使得对这些关键机制的研究变得令人畏惧。因此，不仅需要开发先进的分析方法来测量和识别这些低丰度信号分子，而且还需要应用这些方法来研究更简单的明确系统中的供给回路。该项目的具体目标包括：（1）开发基于质谱的新型策略，以增强原位神经肽分析并首次演示定量直接组织分析；（2）应用这些创新方法来研究神经肽的差异显示饱食且食物匮乏的北方巨蟹（约拿蟹）。开发基于质谱的方法可以对神经肽进行原位定量比较，这将提高采样灵活性并减少组织处理，从而降低分析物损失或污染的风险。甲壳动物神经系统不仅为这项技术的开发和验证提供了一个优秀的测试平台，而且还是研究明确的神经网络中的神经调节的理想模型。其电生理特性易于评估的已识别神经元的高发生率以及丰富的神经调节物质库使这种小型神经网络成为有吸引力的模型系统。该项目具有巨大的潜力，可以发现新的神经肽，并增加对摄食行为肽能调节的神经化学基础的理解。所开发的分析方法不仅适用于更大、更复杂的脊椎动物系统的研究，而且从甲壳类动物系统的研究中获得的分子见解也可能导致高等物种中同源机制的识别。因此，这项研究将改进神经肽分析的方法和扩展能力，并确定开发饮食失调新药物治疗的潜在靶点。 *将开发测量神经系统使用的分子信号的新方法，然后应用于研究如何控制进食行为。这项研究将有助于更好地理解神经系统原理以及针对肥胖等重要健康问题的新疗法。