Abstract
Today, commercially available mass spectrometers increasingly meet all the demands of the proteomics community including high throughput, high sensitivity, and significant fragmentation capability for sequence determinations. Therefore, proper sample preparation is often the most crucial step to obtain the necessary data, particularly when working with biological samples. Depending on the size, sample preparation techniques differ and have to be optimized empirically. This is particularly apparent at the single cell level. In this chapter, we describe protocols for the use of MALDI-TOF mass spectrometry to directly analyse the peptidome of single insect neurons.
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References
Hummon, A.B., Amare, A., and Sweedler, J.V. (2006) Discovering new invertebrate neuropeptides using mass spectrometry. Mass Spectrom. Rev. 25, 77–98.
Rubakhin, S.S. and Sweedler, J.V. (2007) Characterizing peptides in individual mammalian cells using mass spectrometry. Nat. Protoc. 2, 1987–1997.
Neupert, S. and Predel, R. (2005) Mass spectrometric analysis of single identified neurons of an insect. Biochem. Biophys. Res. Commun. 327, 640–645.
Ma, P.W., Garden, R.W., Niermann, J.T., O’Connor, M., Sweedler, J.V., and Roelofs, W.L. (2000) Characterizing the Hez-PBAN gene products in neuronal clusters with immunocytochemistry and MALDI MS. J. Insect Physiol. 46, 221–230.
Neupert, S., Johard, H.A., Nassel, D.R., and Predel, R. (2007) Single-cell peptidomics of Drosophila melanogaster neurons identified by Gal4-driven fluorescence. Anal. Chem. 79, 3690–3694.
Reischig, T. and Stengl, M. (2002) Optic lobe commissures in a three-dimensional brain model of the cockroach Leucophaea maderae: a search for the circadian coupling pathways. J. Comp. Neurol. 443, 388–400.
Brand, A. (1999) GFP as a cell and developmental marker in the Drosophila nervous system. Methods Cell Biol. 58, 165–181.
Nachman, R.J., Russell, W.K., Coast, G.M., Russell, D.H., and Predel, R. (2005) Mass spectrometric assignment of Leu/Ile in neuropeptides from single neurohemal organ preparations of insects. Peptides 26, 2151–2156.
Melcher, C. and Pankratz, M.J. (2005) Candidate gustatory interneurons modulating feeding behavior in the Drosophila brain. PLoS. Biol. 3, e305.
Acknowledgments
Original work was supported by the Deutsche Forschungsgemeinschaft (DFG, PR 595/1-1…7, 6-1…4; and PR 766/9-1 to R Predel, J Schachtner and C Wegener) and by the Peter und Traudl Engelhorn Stiftung (to SN).
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Neupert, S., Predel, R. (2010). Peptidomic Analysis of Single Identified Neurons. In: Soloviev, M. (eds) Peptidomics. Methods in Molecular Biology, vol 615. Humana Press. https://doi.org/10.1007/978-1-60761-535-4_11
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DOI: https://doi.org/10.1007/978-1-60761-535-4_11
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