Two Different Strategies for Stabilization of Brain Tissue and Extraction of Neuropeptides

Elva Fridjonsdottir, Anna Nilsson, Lloyd D. Fricker, Per E. Andrén

Research output: Chapter in Book/Report/Conference proceedingChapter

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 living 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 two different stabilization protocols for rodent brain samples that increase the number of intact mature neuropeptides and minimize interference from degradation products of abundant proteins. Additionally, we present an extraction protocol that aims to extract a wide range of hydrophilic and hydrophobic neuropeptides by sequentially using an aqueous and an organic extraction medium.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages49-60
Number of pages12
DOIs
StatePublished - 2024

Publication series

NameMethods in Molecular Biology
Volume2758
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Brain tissue
  • Electrospray mass spectrometry
  • Extraction
  • Heat inactivation
  • Neuropeptides
  • Postmortem degradation
  • Sample preparation
  • Stabilization

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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