Open AccessAnalysis of microdissected neurons by 18 O mass spectrometry reveals altered protein expression in Alzheimer's disease
Author(s) -
Hashimoto Masakazu,
Bogdanovic Nenad,
Nakagawa Hiroyuki,
Volkmann Inga,
Aoki Mikio,
Winblad Bengt,
Sakai Jun,
Tjernberg Lars O.
Publication year - 2012
Publication title -
journal of cellular and molecular medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.44
H-Index - 130
eISSN - 1582-4934
pISSN - 1582-1838
DOI - 10.1111/j.1582-4934.2011.01441.x
Subject(s) - laser capture microdissection , proteome , western blot , biology , hippocampal formation , proteomics , heat shock protein , immunohistochemistry , hsp70 , hsf1 , alzheimer's disease , microbiology and biotechnology , gene expression , pathology , disease , biochemistry , neuroscience , medicine , gene , immunology
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.