Abstract
Neuropeptides and peptide hormones are involved in the regulation of most if not all body functions, ranging from physiology to neuronal processing and the control of behavior. To assess their functions, it is often vital to determine when and in which quantities they are produced, stored, and released. The latter is especially difficult to assess in small insects, such as the genetically amenable fruit fly Drosophila melanogaster, and cannot be achieved merely by quantifying mRNA transcripts. We have adapted and optimized methods to quantify neuropeptides and peptide hormones by metabolic labeling followed by LC-MS. In this chapter, we describe the labeling protocols used in our laboratory and discuss problems and pitfalls that we encountered.
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Acknowledgments
We thank Gertrud Gramlich for excellent technical help, and Jörg Kahnt (MS facility, Max-Planck-Institute of Terrestrial Microbiology, Marburg, Germany), Jens T. Vanselow, and Andreas Schlosser (MS facility, Virchow Center, University of Würzburg, Germany) for MS measurements and helpful discussions. Original work was supported by the Deutsche Forschungsgemeinschaft (DFG WE 2652/4-2, to CW) and The Wellcome Trust/DBT India Alliance Early Career Fellowship (Award # IA/E/12/1/500742, to Megha).
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Kunz, T.O., Chen, J., Megha, Wegener, C. (2018). Metabolic Labeling to Quantify Drosophila Neuropeptides and Peptide Hormones. In: Schrader, M., Fricker, L. (eds) Peptidomics. Methods in Molecular Biology, vol 1719. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7537-2_11
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DOI: https://doi.org/10.1007/978-1-4939-7537-2_11
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