Abstract
Oxidative stress is a key event in the onset and progression of neurodegenerative diseases, including Alzheimer’s disease (AD). To investigate the role of oxidative stress in AD and to search for potential biomarkers in peripheral blood, serums were collected in this study from the 3-, 6-, and 12-month-old triple transgenic AD mice (3×Tg-AD mice) and the age- and sex-matched non-transgenic (non-Tg) littermates. The serum oxidized proteins were quantified by slot-blot analysis and enzyme-linked immunosorbent assay (ELISA) to investigate the total levels of serum protein carbonyl groups. Western blotting, in conjunction with two-dimensional gel electrophoresis (2D-Oxyblot), was employed to identify and quantify the specifically-carbonylated proteins in the serum of 3×Tg-AD mice. The results showed that the levels of serum protein carbonyls were increased in the three month old 3×Tg-AD mice compared with the non-Tg control mice, whereas no significant differences were observed in the six and 12 months old AD mice, suggesting that oxidative stress is an early event in AD progression. With the application of 2D-Oxyblot analysis, (immunoglobin) Ig gamma-2B chain C region (IGH-3), Ig lambda-2 chain C region (IGLC2), Ig kappa chain C region (IGKC), and Ig kappa chain V-V region HP R16.7 were identified as significantly oxidized proteins compared with the control. Among them IGH-3 and IGKC were validated via immunoprecipitation and Western blot analysis. Identification of oxidized proteins in the serums of 3×Tg-AD mice can not only reveal potential roles of those proteins in the pathogenesis of AD but also provide potential biomarkers of AD at the early stage.
Highlights
Alzheimer’s disease (AD) is the most important cause of dementia in the elderly and is characterized by progressive cognitive deficits that gradually lead to loss of memory and impaired activities in daily living
To study the role that oxidative stress may play in the progression of AD and to search for the biomarkers of AD, we used male 3ˆTg-AD mice and age-matched non-transgenic controls to identify carbonylated serum proteins at early and advanced AD stages (3, 6, and 12-month-old)
We evaluated the levels of total protein carbonyls and identified the oxidative modification proteins in the serums of 3ˆTg-AD mice
Summary
Alzheimer’s disease (AD) is the most important cause of dementia in the elderly and is characterized by progressive cognitive deficits that gradually lead to loss of memory and impaired activities in daily living. A triple transgenic mouse model of AD (3ˆTg-AD) has been developed, which was derived from mixed 129/C57BL6 mice and harboring the mutated genes of APPswe, PS1M146V, and TauP301L [19,20] Those mice exhibit both Aβ and phosphorylated tau alterations, recapitulating most of the pathological features of the AD brains [21]. To study the role that oxidative stress may play in the progression of AD and to search for the biomarkers of AD, we used male 3ˆTg-AD mice and age-matched non-transgenic controls to identify carbonylated serum proteins at early and advanced AD stages (3-, 6-, and 12-month-old). Sroci.u2p01s6,a1n7d, 46t9wo-dimensional gel electrophoresis (2D-Oxyblots) to identify which p3rooft1e7ins were subject to oxidation
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