In Vivo Imaging with Genetically Encoded Redox Biosensors

Kostyuk, Alexander I.; Panova, Anastasiya S.; Kokova, Aleksandra D; Kotova, Daria A.; Maltsev, Dmitry I; Podgorny, Oleg V.; Belousov, Vsevolod V.; Bilan, Dmitry S.

doi:10.3390/ijms21218164

Cited by 41 publications

(19 citation statements)

References 582 publications

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“…In conjunction with measuring stress-related and redox activation signals, it is also important to consider ROS generation via histochemical staining, or quantitative determination through the luminol chemiluminescence assay, and the levels of cell death via electrolyte leakage assays [ 34 , 45 ]. Genetically encoded ROS reporters are increasingly being deployed, to provide spatial–temporal information on subcellular ROS generation [ 46 ]. A range of genetically encoded reporters, with different subcellular locations, capable of measuring ATP [ 47 ], monitoring intracellular H 2 O 2 dynamics [ 48 ], NAD redox status [ 49 ], or dynamic changes in NADPH [ 50 ], are being developed, and these will be invaluable in providing a spatial context to targeted metabolomics approaches.…”

Section: Plant Stress Responses Are Metabolically Diverse and Require A Suite Of Technologies For Accurate Characterizationmentioning

confidence: 99%

Unravelling Plant Responses to Stress—The Importance of Targeted and Untargeted Metabolomics

et al. 2021

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Climate change and an increasing population, present a massive global challenge with respect to environmentally sustainable nutritious food production. Crop yield enhancements, through breeding, are decreasing, whilst agricultural intensification is constrained by emerging, re-emerging, and endemic pests and pathogens, accounting for ~30% of global crop losses, as well as mounting abiotic stress pressures, due to climate change. Metabolomics approaches have previously contributed to our knowledge within the fields of molecular plant pathology and plant–insect interactions. However, these remain incredibly challenging targets, due to the vast diversity in metabolite volatility and polarity, heterogeneous mixtures of pathogen and plant cells, as well as rapid rates of metabolite turn-over. Unravelling the systematic biochemical responses of plants to various individual and combined stresses, involves monitoring signaling compounds, secondary messengers, phytohormones, and defensive and protective chemicals. This demands both targeted and untargeted metabolomics approaches, as well as a range of enzymatic assays, protein assays, and proteomic and transcriptomic technologies. In this review, we focus upon the technical and biological challenges of measuring the metabolome associated with plant stress. We illustrate the challenges, with relevant examples from bacterial and fungal molecular pathologies, plant–insect interactions, and abiotic and combined stress in the environment. We also discuss future prospects from both the perspective of key innovative metabolomic technologies and their deployment in breeding for stress resistance.

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Section: Plant Stress Responses Are Metabolically Diverse and Require A Suite Of Technologies For Accurate Characterizationmentioning

confidence: 99%

Unravelling Plant Responses to Stress—The Importance of Targeted and Untargeted Metabolomics

et al. 2021

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“…Furthermore, a recent study presented redox sensors for the cytosolic glutathione system, the major antioxidant system of the cell [ 127 ]. These genetically encoded redox proteins are very useful for monitoring the redox landscape under experimental conditions [ 128 ]. For instance, these sensors enable the study of molecular pathologies in animal models, such as the oxidative stress-mediated neurodegeneration of motor neurons in zebrafish [ 129 ].…”

Section: Tools and Methods To Monitor The Redox Landscape Of Neuroinflammationmentioning

confidence: 99%

Imaging Biomarkers for Monitoring the Inflammatory Redox Landscape in the Brain

Fernandes

Özcelik

2021

Antioxidants

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Inflammation is one key process in driving cellular redox homeostasis toward oxidative stress, which perpetuates inflammation. In the brain, this interplay results in a vicious cycle of cell death, the loss of neurons, and leakage of the blood–brain barrier. Hence, the neuroinflammatory response fuels the development of acute and chronic inflammatory diseases. Interrogation of the interplay between inflammation, oxidative stress, and cell death in neurological tissue in vivo is very challenging. The complexity of the underlying biological process and the fragility of the brain limit our understanding of the cause and the adequate diagnostics of neuroinflammatory diseases. In recent years, advancements in the development of molecular imaging agents addressed this limitation and enabled imaging of biomarkers of neuroinflammation in the brain. Notable redox biomarkers for imaging with positron emission tomography (PET) tracers are the 18 kDa translocator protein (TSPO) and monoamine oxygenase B (MAO–B). These findings and achievements offer the opportunity for novel diagnostic applications and therapeutic strategies. This review summarizes experimental as well as established pharmaceutical and biotechnological tools for imaging the inflammatory redox landscape in the brain, and provides a glimpse into future applications.

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“…These two probes therefore offer rapid and reversible ratiometric measurement of fluorescence in response to redox changes in reducing compartments, and recombinant roGFP constructs with targeting sequences also allow plant organelle (cytosol, mitochondria, ER, peroxisomes) specific redox sensing. [45][46][47][48][49] These biosensors have been used extensively in plants, for example in examining Arabidopsis responses to drought stress 50 and saline stress 51 (see Kostyuk et al 52 for further examples).…”

Section: Fluorescent Biosensorsmentioning

confidence: 99%