Dataset Information

Ratiometric Imaging of Mitochondrial Hydrogen Peroxide in Aβ₄₂-Mediated Neurotoxicity.

ABSTRACT: Oxidative stress is important for the etiology and pathogenesis of Alzheimer's disease (AD). Research tools that can conveniently evaluate oxidative stress in AD models are expected to catalyze and accelerate research on AD. This study explored the use of genetically encoded fluorescent indicators (GEFIs) to detect mitochondrial oxidative stress in organotypic brain slices and AD mouse models. To enable ratiometric normalization and avoid tissue autofluorescence, we genetically fused a green fluorescent hydrogen peroxide (H₂O₂) indicator, HyPer7, with each of two selected, bright red fluorescent proteins (RFPs), mScarlet-I and tdTomato. The resultant indicators, namely, HyPerGR_S and HyPerGR_T, were tagged with mitochondrial targeting sequences and examined for localization and function in cultured HeLa cells and primary mouse neurons. We further utilized HyPerGR_T, which is a genetic fusion of HyPer7 with tdTomato, to monitor mitochondrial H₂O₂ in response to the human β-amyloid 1-42 isoform (Aβ₄₂) in cultured brain slices and an AD mouse model. Owing to the high sensitivity and low autofluorescence interference resulting from HyPerGR_T, we successfully detected Aβ₄₂-mediated mitochondrial H₂O₂ in these AD models. The results suggest that HyPerGR_T is a valuable tool for studying mitochondrial oxidative stress in tissues and animals.

SUBMITTER: Li X

PROVIDER: S-EPMC8957525 | biostudies-literature | 2022 Mar

REPOSITORIES: biostudies-literature

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Ratiometric Imaging of Mitochondrial Hydrogen Peroxide in Aβ<sub>42</sub>-Mediated Neurotoxicity.

Li Xinyu X Zhang Yiyu Y Ai Hui-Wang HW

ACS sensors 20220228 3

Oxidative stress is important for the etiology and pathogenesis of Alzheimer's disease (AD). Research tools that can conveniently evaluate oxidative stress in AD models are expected to catalyze and accelerate research on AD. This study explored the use of genetically encoded fluorescent indicators (GEFIs) to detect mitochondrial oxidative stress in organotypic brain slices and AD mouse models. To enable ratiometric normalization and avoid tissue autofluorescence, we genetically fused a green flu ...[more]

PMID: 35226474

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Project description:Alzheimer's disease (AD) is a progressive neurodegenerative disorder in which mitochondrial dysfunction and neuroinflammation play crucial roles in its progression. Our previous studies found that cornuside from Cornus officinalis Sieb.Et Zucc is an anti-AD candidate, however, its underlying mechanism remains unknown. In the present study, AD mice were established by intracerebroventricular injection of Aβ1-42 and treated with cornuside (3, 10, 30 mg/kg) for 2 weeks. Cornuside significantly ameliorated behavioral deficits, protected synaptic plasticity and relieved neuronal damage in Aβ1-42 induced mice. Importantly, cornuside decreased NLRP3 inflammasome activation, characterized by decreased levels of NLRP3, ASC, Caspase-1, GSDMD, and IL-1β. Furthermore, cornuside promoted mitophagy accompanied by decreasing SQSTM1/p62 and promoting LC3B-I transforming into LC3B-II, via Pink1/Parkin signaling instead of FUNDC1 or BNIP3 pathways. In order to investigate the relationship between NLRP3 inflammasome and mitophagy in the neuroprotective mechanism of cornuside, we established an in-vitro model in BV2 cells exposed to LPS and Aβ1-42. And cornuside inhibited NLRP3 inflammasome activation and subsequent cytokine release, also protected neurons from damaging factors in microenvironment of conditional culture. Cornuside improved mitochondrial function by promoting oxidative phosphorylation and glycolysis, decreasing the production of ROS and mitochondrial membrane potential depolarization. Besides, mitophagy was also facilitated with increased colocalization of MitoTracker with LC3B and Parkin, and Pink1/Parkin, FUNDC1 and BNIP3 pathways were all involved in the mechanism of cornuside. By blocking the formation of autophagosomes by 3-MA, the protective effects on mitochondria, the inhibition on NLRP3 inflammasome as well as neuronal protection in conditional culture were eliminated. There is reason to believe that the promotion of mitophagy plays a key role in the NLRP3 inhibition of cornuside. In conclusion, cornuside re-establishes the mitophagy flux which eliminates damaged mitochondria and recovers mitochondrial function, both of them are in favor of inhibiting NLRP3 inflammasome activation, then alleviating neuronal and synaptic damage, and finally improving cognitive function.

Dataset Information

Ratiometric Imaging of Mitochondrial Hydrogen Peroxide in Aβ₄₂-Mediated Neurotoxicity.

Publications

Ratiometric Imaging of Mitochondrial Hydrogen Peroxide in Aβ<sub>42</sub>-Mediated Neurotoxicity.

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Dataset Information

Ratiometric Imaging of Mitochondrial Hydrogen Peroxide in Aβ42-Mediated Neurotoxicity.

Publications

Ratiometric Imaging of Mitochondrial Hydrogen Peroxide in Aβ<sub>42</sub>-Mediated Neurotoxicity.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Ratiometric Imaging of Mitochondrial Hydrogen Peroxide in Aβ₄₂-Mediated Neurotoxicity.