A ratiometric fluorescent probe for assessing mitochondrial phospholipid peroxidation within living cells

Prime, Tracy A.; Forkink, Marleen; Logan, Angela; Finichiu, Peter G.; McLachlan, J; Pun, Pamela Boon Li; Koopman, Werner J.H.; Larsen, Lesley; Latter, Melissa J.; Smith, Robin A.J.; Murphy, Michael P.

doi:10.1016/j.freeradbiomed.2012.05.033

Cited by 68 publications

(64 citation statements)

References 37 publications

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“…One hour prior to irradiation, 1 µM C11-BODIPY 581/591 (4,4-difluoro-5-(4-phenyl-1,3-butadienyl)-4-bora-3a,4a-diaza-s-indacene-3-undecanoic acid; ThermoFisher) or 100 nM mito-C11-BODIPY 581/591 (derived from C11-BODIPY 581/591 , conjugated to a triphenylphosphonium lipophilic cation -also known as MitoPerOx, synthesized as previously described [29]) was added to each flask. The triphenylphosphonium conjugation leads to selective uptake of mito-C11-BODIPY 581/591 into mitochondria in cells [29].…”

Section: Detection Of Lipid Peroxidation By C11-bodipymentioning

confidence: 99%

“…The triphenylphosphonium conjugation leads to selective uptake of mito-C11-BODIPY 581/591 into mitochondria in cells [29]. Post-irradiation, cells were washed and re-suspended in 1 ml of cold PBS ready for analysis by flow cytometry.…”

Section: Detection Of Lipid Peroxidation By C11-bodipymentioning

confidence: 99%

“…Lipid peroxidation following photodynamic treatments was assessed using the probe C11-BODIPY 581/591 (Fig. 2F) and its mitochondria-targeted derivative mito-C11-BODIPY 581/591 (MitoPerOx [29], Fig. 2G).…”

Section: Cells Cultured Long-term Under Hyperoxic Conditions Are Resimentioning

confidence: 99%

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Altered cellular redox homeostasis and redox responses under standard oxygen cell culture conditions versus physioxia

Ferguson

Smerdon

Harries

et al. 2018

Free Radical Biology and Medicine

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In vivo, mammalian cells reside in an environment of 0.5-10% O 2 (depending on the tissue location within the body), whilst standard in vitro cell culture is carried out under room air. Little is known about the effects of this hyperoxic environment on treatment-induced oxidative stress, relative to a physiological oxygen environment. In the present study we investigated the effects of long-term culture under hyperoxia (air) on photodynamic treatment. Upon photodynamic irradiation, cells which had been cultured long-term under hyperoxia generated higher concentrations of mitochondrial reactive oxygen species, compared with cells in a physioxic (2% O 2 ) environment. However, there was no significant difference in viability between hyperoxic and physioxic cells. The expression of genes encoding key redox homeostasis proteins and the activity of key antioxidant enzymes was significantly higher after the long-term culture of hyperoxic cells compared with physioxic cells. The induction of antioxidant genes and increased antioxidant enzyme activity appear to contribute to the development of a phenotype that is resistant to oxidative stress-induced cellular damage and death when using standard cell culture conditions. The results from experiments using selective inhibitors suggested that the thioredoxin antioxidant system contributes to this phenotype. To avoid artefactual results, in vitro cellular responses should be studied in mammalian cells that have been cultured under physioxia. This investigation provides new insights into the effects of physioxic cell culture on a model of a clinically relevant photodynamic treatment and the associated cellular pathways.

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Section: Detection Of Lipid Peroxidation By C11-bodipymentioning

confidence: 99%

Section: Detection Of Lipid Peroxidation By C11-bodipymentioning

confidence: 99%

See 1 more Smart Citation

Altered cellular redox homeostasis and redox responses under standard oxygen cell culture conditions versus physioxia

Ferguson

Smerdon

Harries

et al. 2018

Free Radical Biology and Medicine

Self Cite

View full text Add to dashboard Cite

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“…Further, a recently developed mitochondrion-targeted fluorescent lipid peroxidation probe MitoPerOx was shown to report mitochondrial lipid peroxidation in isolated liver cells (284). Highly reactive ROS, including OH, peroxynitrite (ONOO -), and hypochlorous acid (HOCl), can be measured in mitochondria by rhodamine-like fluorophores capped with either a 4-aminophenyl aryl ether (MitoAR) or a 4-hydroxy aryl ether (MitoHR) group (187).…”

Section: Compartment-specific Ros Measurementmentioning

confidence: 99%

Nutritional Countermeasures Targeting Reactive Oxygen Species in Cancer: From Mechanisms to Biomarkers and Clinical Evidence

Samoylenko

Hossain²,

Mennerich³

et al. 2013

Antioxidants & Redox Signaling

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Reactive oxygen species (ROS) exert various biological effects and contribute to signaling events during physiological and pathological processes. Enhanced levels of ROS are highly associated with different tumors, a Western lifestyle, and a nutritional regime. The supplementation of food with traditional antioxidants was shown to be protective against cancer in a number of studies both in vitro and in vivo. However, recent largescale human trials in well-nourished populations did not confirm the beneficial role of antioxidants in cancer, whereas there is a well-established connection between longevity of several human populations and increased amount of antioxidants in their diets. Although our knowledge about ROS generators, ROS scavengers, and ROS signaling has improved, the knowledge about the direct link between nutrition, ROS levels, and cancer is limited. These limitations are partly due to lack of standardized reliable ROS measurement methods, easily usable biomarkers, knowledge of ROS action in cellular compartments, and individual genetic predispositions. The current review summarizes ROS formation due to nutrition with respect to macronutrients and antioxidant micronutrients in the context of cancer and discusses signaling mechanisms, used biomarkers, and its limitations along with large-scale human trials.

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“…Next to direct ROS probes, one could also use probes that assess ROS indirectly by measuring the downstream damage such as lipid peroxidation. C11-BODIPY 581/591 , for example, is a ratiometric sensor for lipid peroxidation (Drummen et al 2002), while MitoPeroX (a mitochondria targeted derivative of C11-BODIPY 581/591 ) is a ratiometric probe for the specific assessment of mitochondrial phospholipid peroxidation (Prime et al 2012). Several fluorescent dyes have also been developed for measuring mitochondrial morphology and Δψ m .…”

Section: Sensorsmentioning

confidence: 99%

Integrated High-Content Quantification of Intracellular ROS Levels and Mitochondrial Morphofunction

Sieprath

Corne

Willems

et al. 2016

Advances in Anatomy, Embryology and Cell Biology

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Oxidative stress arises from an imbalance between the production of reactive oxygen species (ROS) and their removal by cellular antioxidant systems. Especially under pathological conditions, mitochondria constitute a relevant source of cellular ROS. These organelles harbor the electron transport chain, bringing electrons in close vicinity to molecular oxygen. Although a full understanding is still lacking, intracellular ROS generation and mitochondrial function are also linked to changes in mitochondrial morphology. To study the intricate relationships between the different factors that govern cellular redox balance in living cells, we have developed a high-content microscopy-based strategy for simultaneous quantification of intracellular ROS levels and mitochondrial morphofunction. Here, we summarize the principles of intracellular ROS generation and removal, and we explain the major considerations for performing quantitative microscopy analyses of ROS and mitochondrial morphofunction in living cells. Next, we describe our workflow and finally, we illustrate that a multiparametric readout enables the unambiguous classification of chemically perturbed cells as well as laminopathy patient cells. Principles of intracellular ROS generation and removalReactive oxygen species (ROS) are small, short-lived derivatives of molecular oxygen (O 2 ) of radical and non-radical nature (Halliwell and Gutteridge 2007 • ), nitrate radical (NO 3 • ) and many other nitrogen derivatives. ROS were originally described as molecular constituents of the defense system of phagocytic cells, but it has become clear that besides their damaging properties, they also function as signaling molecules and mediate a variety of other cellular responses including cell proliferation, differentiation, gene expression and migration (Lambeth 2004;Bartz and Piantadosi 2010). Intracellular ROS metabolismROS can be generated at various sites in the cell (Fig. 1A). This can be either deliberately, e.g. by NADPH oxidases (NOX), or as a byproduct, e.g. during normal cellular respiration in mitochondria (Babior 1999;Turrens 2003;Murphy 2009). The NOX family of NADPH oxidases (NOX1, NOX2, NOX3, NOX4, NOX5, DUOX1 and DUOX2) are proteins that transport electrons (e -) from NADPH across biological membranes (plasma or endomembranes) (Bedard and Krause 2007;Dupre-Crochet et al. 2013). The activation mechanisms and tissue distribution of the isoforms differ but they all use O 2 as e --acceptor, producing O 2•-. Through ROS generation, they play a role in many cellular processes including host defense, regulation of gene expression and cell differentiation (Bedard and Krause 2007). Despite their sometimes significant contribution to the global ROS pools, NOX are not the predominant source of intracellular ROS. Mitochondria are considered the major culprit, in particular under pathological conditions. Mitochondrial ROS are generated as a byproduct of the oxidative phosphorylation (OXPHOS, cf. below). Irrespective of its source, ROS production generally sta...

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A ratiometric fluorescent probe for assessing mitochondrial phospholipid peroxidation within living cells

Cited by 68 publications

References 37 publications

Altered cellular redox homeostasis and redox responses under standard oxygen cell culture conditions versus physioxia

Altered cellular redox homeostasis and redox responses under standard oxygen cell culture conditions versus physioxia

Nutritional Countermeasures Targeting Reactive Oxygen Species in Cancer: From Mechanisms to Biomarkers and Clinical Evidence

Integrated High-Content Quantification of Intracellular ROS Levels and Mitochondrial Morphofunction

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