Evaluation of Mitochondrial Oxidative Stress in the Brain of a Transgenic Mouse Model of Alzheimer’s Disease by in vitro Electron Paramagnetic Resonance Spectroscopy

Article type: Research Article

Authors: Manabe, Tatsuo^{a; 1} | Matsumura, Akihiro^{a; 1} | Yokokawa, Kazuki^a | Saito, Taro^a | Fujikura, Mai^a | Iwahara, Naotoshi^a | Matsushita, Takashi^a | Suzuki, Syuuichirou^a | Hisahara, Shin^a | Kawamata, Jun^a | Suzuki, Hiromi^a | Emoto, Miho C.^b | Fujii, Hirotada G.^b | Shimohama, Shun^{a; *}

Affiliations: [a] Department of Neurology, School of Medicine, Sapporo Medical University, Sapporo, Japan | [b] Health Sciences University of Hokkaido, Sapporo, Japan

Correspondence: [*] Correspondence to: Dr. Shun Shimohama, Department of Neurology, School of Medicine, Sapporo Medical University, South 1, West 16, Chuo-ku, Sapporo 060-8543, Japan. Tel.: +81-11-611-2111; Fax: +81-11-622-7668; E-mail: [email protected].

Note: [1] These authors contributed equally to this work.

Abstract: Alzheimer’s disease (AD) is one of the most common neurodegenerative diseases responsible for progressive dementia. Deposition of amyloid-β (Aβ) in the brain is the most important pathophysiological hallmark of AD. In addition, recent evidence indicates that reactive oxygen species (ROS) derived from mitochondria contribute to progression of AD pathology. We thus hypothesized that Aβ accumulates and oxidative stress increases in the brain mitochondria of a transgenic mouse model of AD (APdE9). We measured the quantity of Aβ and the activity of the antioxidant enzyme superoxide dismutase (SOD) in brain mitochondrial fractions prepared from APdE9 and wild-type (WT) mice aged 6, 9, 15, and 18 months. We also quantified the age-related changes in redox status in the mitochondrial fractions obtained from both APdE9 and WT mouse brains by electron paramagnetic resonance (EPR) spectrometry using a paramagnetic nitroxide “Mito-Tempo” [(2-(2,2,6,6-Tetramethylpiperidin-1-oxyl-4-ylamino)-2-oxoethyl) triphenylphosphonium chloride monohydrate] as a mitochondria-targeted redox-sensitive probe. In APdE9 mice, Aβ accumulated in brain mitochondria earlier than in the non-mitochondrial fraction of the brain. Furthermore, increased oxidative stress was demonstrated in brain mitochondria of APdE9 mice by in vitro SOD assay as well as EPR spectroscopy. EPR combined with a mitochondria-targeted redox-sensitive nitroxide probe is a potentially powerful tool to elucidate the etiology of AD and facilitate the development of new therapeutic strategies for AD.

Keywords: Alzheimer’s disease, amyloid-β, electron paramagnetic resonance, mitochondria, reactive oxygen special, redox status, oxidative stress

DOI: 10.3233/JAD-180985

Journal: Journal of Alzheimer's Disease, vol. 67, no. 3, pp. 1079-1087, 2019