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利用质谱法鉴定和靶向定量线虫秀丽隐杆线虫中的内源性神经肽。

Identification and Targeted Quantification of Endogenous Neuropeptides in the Nematode Caenorhabditis elegans Using Mass Spectrometry.

机构信息

Department of Biology, Animal Physiology & Neurobiology, University of Leuven (KU Leuven), Leuven, Belgium.

Bavarian Center for Biomolecular Mass Spectrometry (BayBioMS), Technical University of Munich (TUM), Freising, Germany.

出版信息

Methods Mol Biol. 2024;2758:341-373. doi: 10.1007/978-1-0716-3646-6_19.

Abstract

The nematode Caenorhabditis elegans lends itself as an excellent model organism for peptidomics studies. Its ease of cultivation and quick generation time make it suitable for high-throughput studies. The nervous system, with its 302 neurons, is probably the best-known and studied endocrine tissue. Moreover, its neuropeptidergic signaling pathways display numerous similarities with those observed in other metazoans. Here, we describe two label-free approaches for neuropeptidomics in C. elegans: one for discovery purposes, and another for targeted quantification and comparisons of neuropeptide levels between different samples. Starting from a detailed peptide extraction procedure, we here outline the liquid chromatography tandem mass spectrometry (LC-MS/MS) setup and describe subsequent data analysis approaches.

摘要

秀丽隐杆线虫是一种优秀的肽组学研究模式生物。它易于培养且世代时间短,非常适合进行高通量研究。其神经系统拥有 302 个神经元,可能是最知名和研究最多的内分泌组织。此外,其神经肽信号通路与其他后生动物中观察到的信号通路具有许多相似之处。在这里,我们描述了秀丽隐杆线虫中两种非标记肽组学方法:一种用于发现目的,另一种用于靶向定量和比较不同样品之间的神经肽水平。我们从详细的肽提取程序开始,在此概述了液相色谱串联质谱(LC-MS/MS)设置,并描述了随后的数据分析方法。

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