Proteomic and functional alterations in brain mitochondria from Tg2576 mice occur before amyloid plaque deposition.
Synaptic dysfunction is an early event in Alzheimer's disease patients and has also been detected in transgenic mouse models. In the present study, we analyzed proteomic changes in synaptosomal fractions from Tg2576 mice that overexpress mutant human amyloid precursor protein (K670N, M671L) and from their nontransgenic littermates. Cortical and hippocampal tissue was microdissected at the onset of cognitive impairment, but before deposition of amyloid plaques. Crude synaptosomal fractions were prepared by differential centrifugation, proteins were separated by 2-D DIGE and identified by MS/MS. Significant alterations were detected in mitochondrial heat shock protein 70 pointing to a mitochondrial stress response. Subsequently, synaptosomal versus nonsynaptic mitochondria were purified from Tg2576 mice brains by density gradient centrifugation. Mitochondrial proteins were separated by IEF or Blue-native gel electrophoresis in the first dimension and SDS-PAGE in the second dimension. Numerous changes in the protein subunit composition of the respiratory chain complexes I and III were identified. Levels of corresponding mRNAs remain unchanged as shown by Affymetrix oligonucleotide array analysis. Functional examination revealed impaired state 3 respiration and uncoupled respiration in brain mitochondria from young Tg2576 mice. By immunoblotting, amyloid-beta oligomers were detected in synaptosomal fractions from Tg2576 mice and reduced glucose metabolism was observed in Tg2576 mice brains by [14C]-2-deoxyglucose infusion. Taken together, we demonstrate alterations in the mitochondrial proteome and function that occur in Tg2576 mice brains before amyloid plaque deposition suggesting that mitochondria are early targets of amyloid-beta aggregates.
Gillardon
Frank
F
Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, Germany.
Rist
Wolfgang
W
Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, Germany.
Kussmaul
Lothar
L
Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, Germany.
Vogel
Johannes
J
University of Zurich, Zurich, Switzerland.
Berg
Matthias
M
GE Healthcare, Munich, Germany.
Danzer
Karin
K
Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, Germany.
Kraut
Norbert
N
Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, Germany.
Hengerer
Bastian
B
Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, Germany.
D000682:Amyloid / Q000378:metabolism*; D016564:Amyloid beta-Protein Precursor / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D001921:Brain / Q000378:metabolism* / Q000473:pathology; D002250:Carbon Radioisotopes / Q000737:chemistry; D005947:Glucose / Q000378:metabolism; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D008928:Mitochondria / Q000378:metabolism*; D020543:Proteome / Q000378:metabolism*; D013574:Synaptosomes / Q000378:metabolism*; D053719:Tandem Mass Spectrometry
D000682:Amyloid; D016564:Amyloid beta-Protein Precursor; D002250:Carbon Radioisotopes; D020543:Proteome; D005947:Glucose