Department of Chemistry, University of Wisconsin-Madison, Madison, WI, USA.
School of Pharmacy, University of Wisconsin-Madison, Madison, WI, USA.
Methods Mol Biol. 2024;2758:255-289. doi: 10.1007/978-1-0716-3646-6_14.
Crustaceans serve as a useful, simplified model for studying peptides and neuromodulation, as they contain numerous neuropeptide homologs to mammals and enable electrophysiological studies at the single-cell and neural circuit levels. Crustaceans contain well-defined neural networks, including the stomatogastric ganglion, oesophageal ganglion, commissural ganglia, and several neuropeptide-rich organs such as the brain, pericardial organs, and sinus glands. As existing mass spectrometry (MS) methods are not readily amenable to neuropeptide studies, there is a great need for optimized sample preparation, data acquisition, and data analysis methods. Herein, we present a general workflow and detailed methods for MS-based neuropeptidomic analysis of crustacean tissue samples and circulating fluids. In conjunction with profiling, quantitation can also be performed with isotopic or isobaric labeling. Information regarding the localization patterns and changes of peptides can be studied via mass spectrometry imaging. Combining these sample preparation strategies and MS analytical techniques allows for a multi-faceted approach to obtaining deep knowledge of crustacean peptidergic signaling pathways.
甲壳类动物作为研究肽和神经调节的有用简化模型，因为它们含有许多与哺乳动物同源的神经肽，并能够在单细胞和神经回路水平上进行电生理研究。甲壳类动物含有明确的神经网络，包括口胃神经节、食管神经节、交感神经节以及几个富含神经肽的器官，如脑、心包器官和窦腺。由于现有的质谱 (MS) 方法不适用于神经肽研究，因此非常需要优化的样品制备、数据采集和数据分析方法。本文介绍了一种基于 MS 的甲壳类动物组织样品和循环液中神经肽组学分析的通用工作流程和详细方法。与分析物鉴定相结合，还可以使用同位素或等压标记进行定量。通过质谱成像可以研究肽的定位模式和变化信息。通过组合这些样品制备策略和 MS 分析技术，可以采用多方面的方法来深入了解甲壳类动物的肽信号通路。
