Identification, Quantitation, and Imaging of the Crustacean Peptidome
Identification, Quantitation, and Imaging of the Crustacean Peptidome
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DOI：
10.1007/978-1-4939-7537-2_17
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发表时间：
2018-01-01
期刊：
影响因子：
--
通讯作者：
Li, Lingjun
中科院分区：
文献类型：
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作者：
DeLaney, Kellen;Buchberger, Amanda;Li, Lingjun
甲壳类动物作为一个有用的，简化的模型，研究肽和神经调节，因为它们含有大量的神经肽同源哺乳动物，使电生理学研究在单细胞和神经回路水平。特别是，甲壳类动物包含明确的神经网络，包括口胃神经节，食道神经节，连合神经节和几个富含神经肽的器官，如大脑，心包器官和窦腺。由于缺乏甲壳动物肽的基因组数据库，甲壳动物肽组学的一个重要步骤涉及新肽的发现和鉴定以及数据库的构建，最近借助于质谱（MS）。在这里，我们提出了一个一般的工作流程和详细的方法，甲壳动物组织样品和循环液的MS为基础的肽组分析。结合分析，定量也可以用同位素或同量异位素标记进行。关于肽的定位模式和变化的信息可以通过质谱成像来研究。结合这些样品制备策略和MS分析技术允许多方面的方法来获得甲壳动物肽能信号通路的深入了解。
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. In particular, crustaceans contain well-defined neural networks, including the stomatogastric ganglion, esophageal ganglion, commissural ganglia, and several neuropeptide-rich organs, such as the brain, pericardial organs, and sinus glands. Due to the lack of a genomic database for crustacean peptides, an important step of crustacean peptidomics involves the discovery and identification of novel peptides and the construction of a database, more recently with the aid of mass spectrometry (MS). Herein, we present a general workflow and detailed methods for MS-based peptidomic 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 multifaceted approach to obtaining deep knowledge of crustacean peptidergic signaling pathways.