Redox imaging using genetically encoded redox indicators in zebrafish and mice

Breckwoldt, Michael O.; Wittmann, Christine; Misgeld, Thomas; Kerschensteiner, Martin; Grabher, Clemens

doi:10.1515/hsz-2014-0294

Cited by 16 publications

(9 citation statements)

References 68 publications

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“…We have recently characterized a physiological redox signal in neuronal mitochondria that goes along with a profound shape change of the organelle (dubbed “mitochondrial contractions”) 8 27 . In this first description, mitochondrial redox signals were only investigated at a single organelle level.…”

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confidence: 99%

Mitochondrial redox and pH signaling occurs in axonal and synaptic organelle clusters

Breckwoldt

Armoundas

Aon

et al. 2016

Sci Rep

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Redox switches are important mediators in neoplastic, cardiovascular and neurological disorders. We recently identified spontaneous redox signals in neurons at the single mitochondrion level where transients of glutathione oxidation go along with shortening and re-elongation of the organelle. We now have developed advanced image and signal-processing methods to re-assess and extend previously obtained data. Here we analyze redox and pH signals of entire mitochondrial populations. In total, we quantified the effects of 628 redox and pH events in 1797 mitochondria from intercostal axons and neuromuscular synapses using optical sensors (mito-Grx1-roGFP2; mito-SypHer). We show that neuronal mitochondria can undergo multiple redox cycles exhibiting markedly different signal characteristics compared to single redox events. Redox and pH events occur more often in mitochondrial clusters (medium cluster size: 34.1 ± 4.8 μm2). Local clusters possess higher mitochondrial densities than the rest of the axon, suggesting morphological and functional inter-mitochondrial coupling. We find that cluster formation is redox sensitive and can be blocked by the antioxidant MitoQ. In a nerve crush paradigm, mitochondrial clusters form sequentially adjacent to the lesion site and oxidation spreads between mitochondria. Our methodology combines optical bioenergetics and advanced signal processing and allows quantitative assessment of entire mitochondrial populations.

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confidence: 99%

Mitochondrial redox and pH signaling occurs in axonal and synaptic organelle clusters

Breckwoldt

Armoundas

Aon

et al. 2016

Sci Rep

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“…Genetically encoded biosensors based on FPs. For more than 10 years, genetically encoded biosensors based on FPs have been developed for redox imaging in cell culture and R671 BIOSENSORS FOR ROS DETECTION tissues but also in vivo (20,45,131). One of their main advantages is their biocompatibility as they can be stably expressed in transgenic animals and targeted to a specific location.…”

Section: Toward In Vivo Ros Detection: From Experimental Setup To Chomentioning

confidence: 99%

“…Genetically encoded ROS sensors (cp-YFP, HyPer, or roGFP2-Orp1) were successfully expressed in transgenic organisms including mouse, zebrafish, Xenopus, Caenorhabditis elegans, and plants to study the ROS production in situ (14,20,56). In particular, it was possible to monitor H 2 O 2 levels during development of transgenic zebrafish (56).…”

Section: R672mentioning

confidence: 99%

Biosensors for spatiotemporal detection of reactive oxygen species in cells and tissues

Erard

Dupré-Crochet

Nüße

2018

American Journal of Physiology-Regulatory, Integrative and Comp

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Redox biology has become a major issue in numerous areas of physiology. Reactive oxygen species (ROS) have a broad range of roles from signal transduction to growth control and cell death. To understand the nature of these roles, accurate measurement of the reactive compounds is required. An increasing number of tools for ROS detection is available; however, the specificity and sensitivity of these tools are often insufficient. Furthermore, their specificity has been rarely evaluated in complex physiological conditions. Many ROS probes are sensitive to environmental conditions in particular pH, which may interfere with ROS detection and cause misleading results. Accurate detection of ROS in physiology and pathophysiology faces additional challenges concerning the precise localization of the ROS and the timing of their production and disappearance. Certain ROS are membrane permeable, and certain ROS probes move across cells and organelles. Targetable ROS probes such as fluorescent protein-based biosensors are required for accurate localization. Here we analyze these challenges in more detail, provide indications on the strength and weakness of current tools for ROS detection, and point out developments that will provide improved ROS detection methods in the future. There is no universal method that fits all situations in physiology and cell biology. A detailed knowledge of the ROS probes is required to choose the appropriate method for a given biological problem. The knowledge of the shortcomings of these probes should also guide the development of new sensors.

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“…Therefore, optical imaging combined with the use of fluorescent indicators is a fundamental tool used in basic research; this method enables the noninvasive detection of various intracellular metabolites or signaling molecules in real time with single-cell or even subcellular resolution. Genetically encoded biosensors have many advantages over other types of fluorescent probes (reviewed in [ 20 , 21 , 23 , 24 , 25 , 26 ]) and have allowed unmatched opportunities for studying hypoxic/ischemic injury of the brain. Genetically encoded biosensors may be expressed in a spatiotemporally defined manner in specific cell types or subcellular compartments.…”

Section: Introductionmentioning

confidence: 99%

Genetically Encoded Tools for Research of Cell Signaling and Metabolism under Brain Hypoxia

et al. 2020

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Hypoxia is characterized by low oxygen content in the tissues. The central nervous system (CNS) is highly vulnerable to a lack of oxygen. Prolonged hypoxia leads to the death of brain cells, which underlies the development of many pathological conditions. Despite the relevance of the topic, different approaches used to study the molecular mechanisms of hypoxia have many limitations. One promising lead is the use of various genetically encoded tools that allow for the observation of intracellular parameters in living systems. In the first part of this review, we provide the classification of oxygen/hypoxia reporters as well as describe other genetically encoded reporters for various metabolic and redox parameters that could be implemented in hypoxia studies. In the second part, we discuss the advantages and disadvantages of the primary hypoxia model systems and highlight inspiring examples of research in which these experimental settings were combined with genetically encoded reporters.

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Redox imaging using genetically encoded redox indicators in zebrafish and mice

Cited by 16 publications

References 68 publications

Mitochondrial redox and pH signaling occurs in axonal and synaptic organelle clusters

Mitochondrial redox and pH signaling occurs in axonal and synaptic organelle clusters

Biosensors for spatiotemporal detection of reactive oxygen species in cells and tissues

Genetically Encoded Tools for Research of Cell Signaling and Metabolism under Brain Hypoxia

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