Alterations of brain and cerebellar proteomes linked to Aβ and tau pathology in a female triple-transgenic murine model of Alzheimer's disease

D Ciavardelli,A Del Viscovo,S L Sensi,M Moreno,D De Gregorio,C Di Ilio,E Silvestri,F Goglia,L M T Canzoniero,M Bomba

doi:10.1038/cddis.2010.68

D Ciavardelli, A Del Viscovo + Show 8 more

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https://doi.org/10.1038/cddis.2010.68

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Abstract

The triple-transgenic Alzheimer (3 × Tg-AD) mouse expresses mutant PS1M146V, APPswe, and tauP301L transgenes and progressively develops plaques and neurofibrillary tangles with a temporal- and region-specific profile that resembles the neuropathological progression of Alzheimer's disease (AD). In this study, we used proteomic approaches such as two-dimensional gel electrophoresis and mass spectrometry to investigate the alterations in protein expression occurring in the brain and cerebellum of 3 × Tg-AD and presenilin-1 (PS1) knock-in mice (animals that do not develop Aβ- or tau-dependent pathology nor cognitive decline and were used as control). Finally, using the Ingenuity Pathway Analysis we evaluated novel networks and molecular pathways involved in this AD model. We identified several differentially expressed spots and analysis of 3 × Tg-AD brains showed a significant downregulation of synaptic proteins that are involved in neurotransmitter synthesis, storage and release, as well as a set of proteins that are associated with cytoskeleton assembly and energy metabolism. Interestingly, in the cerebellum, a structure not affected by AD, we found an upregulation of proteins involved in carbohydrate metabolism and protein catabolism. Our findings help to unravel the pathogenic brain mechanisms set in motion by mutant amyloid precursor protein (APP) and hyperphosphorylated tau. These data also reveal cerebellar pathways that may be important to counteract the pathogenic actions of Aβ and tau, and ultimately offer novel targets for therapeutic intervention.

Highlights

presenilin-1 knock-in (PS1-KI) mice harboring the Familial AD (FAD)-related M146V PS1 mutation do not express mutant amyloid precursor protein (APP) or h-tau, do not show any Alzheimer’s disease (AD)-related cognitive decline nor Ab- or tau-mediated pathology,[9] thereby providing an adequate control to investigate the specific impact of the AD-like Ab and tau pathology occurring in 3ÂTg-AD mice
Protein expression profile and identification of differentially expressed proteins in the brain and cerebellum of PS1-KI and 3ÂTg-AD mice. 2D-E-based analysis of protein expression profile of brain and cerebellum in PS1-KI (CTRL) and 3 Â Tg-AD mice showed that 2D-E gel protein-spot patterns were qualitatively and quantitatively similar across all the analyzed gels
In our study, using a classical proteomic approach based on 2-DE and mass spectrometry (MS), we analyzed the differential protein expression associated with the combination of mutant PS1M146V, APPSwe, and tauP301L in the brain and cerebellum of 14-month-old 3ÂTg-AD female mice

Summary

Introduction

Evolutionary Relationship; IPA, Ingenuity Pathway Analysis; DTT, dithiothreitol; SDS-PAGE, sodium dodecyl sulphate-polyacrylamide gel electrophoresis; IAA, iodoacetamide; MALDI-TOF-MS, matrix-assisted laser desorption ionization-time of flight MS; HR, hit ratio; ELDP, excess of limit-digested peptides; MC, mass coverage; PMFscore, peptide mass fingerprint score; PSD, post source decay; ARHGDA, rho-GDP dissociation inhibitor 1; CKB, brain creatine kinase; DPYSL2, dihydropyrimidinase-like[2] protein; ENO1, enolase 1 (a); GLUD1, glutamate dehydrogenase 1; IDH3A, isocitrate dehydrogenase 3 (NAD þ ) a; LDHB, lactate dehydrogenase B; ACO2, mitochondrial aconitase 2; GPD2, mitochondrial glycerol-3-phosphate dehydrogenase 2; OGDHL, oxoglutarate dehydrogenase-like protein; UQCRC1, ubiquinol-cytochrome c reductase core protein I; DNM1, dynamin 1; |NEFM, neurofilament, medium polypeptide; PITPNA, phosphatidylinositol transfer protein, a; SNAP25, synaptosomal-associated protein, 25 kDa; SNCA, synuclein-a; TPI1, triosephosphate isomerase 1; TPM1, tropomyosin 1; ATP6V1A, ATPase, H þ transporting, lysosomal 70 kDa, V1 subunit A; WDR1, WD repeat domain 1; CA2, carbonic anhydrase II; CCT5, chaperonin-containing TCP1subunit 5. The link between tau protein mutations and NFT formation in the AD brain is still unknown; mutant P301L tau leads to its hyperphosphorylation and NFT production in frontotemporal dementia as well as in a form of parkinsonism linked to a mutation found in chromosome 17.4 not specific for AD, the P301L tau mutation is key to model tau neuropathology in transgenic animal models. Several studies have described the differential protein expression profile in the brain of transgenic AD models (reviewed in Sowell et al.6) and a recent study has analyzed protein alterations occurring in male 3ÂTg-AD mice.[7] In contrast, the cerebellar proteome in samples from AD mice or AD patients has been less investigated, most likely because the prevailing assumption is that this central nervous system (CNS) area is relatively unaffected by AD. Using the Protein Analysis Through Evolutionary Relationship (PANTHER) application system and Ingenuity Pathway Analysis (IPA), we inferred the functional role and networks linked to the proteins that we found differentially expressed

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