Preparation of Drosophila Larval Samples for Gas Chromatography-Mass Spectrometry (GC-MS)-based Metabolomics
Preparation of Drosophila Larval Samples for Gas Chromatography-Mass Spectrometry (GC-MS)-based Metabolomics
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DOI：
10.3791/57847
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发表时间：
2018-06-01
影响因子：
1.2
通讯作者：
Tennessen, Jason M.
中科院分区：
文献类型：
--
作者：
Li, Hongde;Tennessen, Jason M.
代谢组学领域的最新进展使果蝇黑腹果蝇成为研究动物代谢的强大遗传模型。通过将大量果蝇遗传工具与调查大量中间代谢的能力相结合，代谢组学方法可以揭示饮食，基因型，生活史事件和环境线索之间的复杂相互作用。此外，代谢组学研究可以发现新的酶机制，并发现以前未知的看似不同的代谢途径之间的联系。为了促进更广泛地使用这种技术在果蝇社区，在这里，我们提供了一个详细的协议，描述了如何准备果蝇幼虫样品的气相色谱-质谱（GC-MS）为基础的代谢组学分析。我们的方案包括幼虫样品收集，代谢物提取，化学衍生化和GC-MS分析的描述。成功完成该方案将允许用户测量小极性代谢物的相对丰度，包括参与糖酵解和TCA循环的氨基酸、糖和有机酸。
Recent advances in the field of metabolomics have established the fruit fly Drosophila melanogaster as a powerful genetic model for studying animal metabolism. By combining the vast array of Drosophila genetic tools with the ability to survey large swaths of intermediary metabolism, a metabolomics approach can reveal complex interactions between diet, genotype, life-history events, and environmental cues. In addition, metabolomics studies can discover novel enzymatic mechanisms and uncover previously unknown connections between seemingly disparate metabolic pathways. In order to facilitate more widespread use of this technology among the Drosophila community, here we provide a detailed protocol that describes how to prepare Drosophila larval samples for gas chromatography-mass spectrometry (GC-MS)-based metabolomic analysis. Our protocol includes descriptions of larval sample collection, metabolite extraction, chemical derivatization, and GC-MS analysis. Successful completion of this protocol will allow users to measure the relative abundance of small polar metabolites, including amino acids, sugars, and organic acids involved in glycolysis and the TCA cycles.