Analysis of microdissected neurons by 18O mass spectrometry reveals altered protein expression in Alzheimer's disease

J Cell Mol Med. 2012 Aug;16(8):1686-700. doi: 10.1111/j.1582-4934.2011.01441.x.

Abstract

It is evident that the symptoms of Alzheimer's disease (AD) are derived from severe neuronal damage, and especially pyramidal neurons in the hippocampus are affected pathologically. Here, we analysed the proteome of hippocampal neurons, isolated from post-mortem brains by laser capture microdissection. By using (18)O labelling and mass spectrometry, the relative expression levels of 150 proteins in AD and controls were estimated. Many of the identified proteins are involved in transcription and nucleotide binding, glycolysis, heat-shock response, microtubule stabilization, axonal transport or inflammation. The proteins showing the most altered expression in AD were selected for immunohistochemical analysis. These analyses confirmed the altered expression levels, and showed in many AD cases a pathological pattern. For comparison, we also analysed hippocampal sections by Western blot. The expression levels found by this method showed poor correlation with the neuron-specific analysis. Hence, we conclude that cell-specific proteome analysis reveals differences in the proteome that cannot be detected by bulk analysis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aged
  • Aged, 80 and over
  • Alzheimer Disease / metabolism*
  • Alzheimer Disease / pathology*
  • Amino Acid Sequence
  • Blotting, Western
  • Case-Control Studies
  • Demography
  • Down-Regulation
  • Female
  • Hippocampus / metabolism
  • Hippocampus / pathology
  • Humans
  • Immunohistochemistry
  • Isotope Labeling
  • Laser Capture Microdissection*
  • Male
  • Mass Spectrometry / methods*
  • Microglia / metabolism
  • Molecular Sequence Data
  • Nerve Tissue Proteins / metabolism*
  • Neurons / metabolism*
  • Neurons / pathology*
  • Oxygen Isotopes
  • Peptides / chemistry
  • Peptides / metabolism
  • Proteome / metabolism
  • Pyramidal Cells / metabolism
  • Pyramidal Cells / pathology
  • Trypsin / metabolism
  • Up-Regulation

Substances

  • Nerve Tissue Proteins
  • Oxygen Isotopes
  • Peptides
  • Proteome
  • Trypsin