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Brain Tissue Sample Stabilization and Extraction Strategies for Neuropeptidomics

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Peptidomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1719))

Abstract

Neuropeptides are bioactive peptides that are synthesized and secreted by neurons in signaling pathways in the brain. Peptides and proteins are extremely vulnerable to proteolytic cleavage when their biological surrounding changes. This makes neuropeptidomics challenging due to the rapid alterations that occur to the peptidome after harvesting of brain tissue samples. For a successful neuropeptidomic study the biological tissue sample analyzed should resemble the premortem state as much as possible. Heat stabilization has been proven to inhibit postmortem degradation by denaturing proteolytic enzymes, hence increasing identification rates of neuropeptides. Here, we describe a stabilization protocol of a frozen tissue specimen that increases the number of intact mature neuropeptides identified and minimizes interference of degradation products from abundant proteins. Additionally, we present an extraction protocol that aims to extract a wide range of hydrophilic and hydrophobic neuropeptides by using both an aqueous and an organic extraction medium.

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Author information

Authors and Affiliations

  1. Biomolecular Imaging and Proteomics, Department of Pharmaceutical Biosciences, Uppsala University, Uppsala, Sweden

    Elva Fridjonsdottir, Anna Nilsson, Henrik Wadensten & Per E. Andrén

Authors
  1. Elva Fridjonsdottir
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  2. Anna Nilsson
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  3. Henrik Wadensten
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  4. Per E. Andrén
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Corresponding author

Correspondence to Per E. Andrén .

Editor information

Editors and Affiliations

  1. Department of Biotechnology and Bioinformatics, Weihenstephan-Tr. University of Applied Sciences, Freising, Bayern, Germany

    Michael Schrader

  2. Department of Molecular Pharmacology, Albert Einstein College of Medicine, Bronx, New York, USA

    Lloyd Fricker

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Fridjonsdottir, E., Nilsson, A., Wadensten, H., Andrén, P.E. (2018). Brain Tissue Sample Stabilization and Extraction Strategies for Neuropeptidomics. 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_2

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  • DOI: https://doi.org/10.1007/978-1-4939-7537-2_2

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7536-5

  • Online ISBN: 978-1-4939-7537-2

  • eBook Packages: Springer Protocols

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