Fluorescence and chemiluminescence approaches for peroxynitrite detection

Prolo, Carolina; Ríos, Natalia; Piacenza, Lucı́a; Álvarez, María Noel; Radí, Rafael

doi:10.1016/j.freeradbiomed.2018.02.017

Cited by 81 publications

(44 citation statements)

References 111 publications

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“…Free radicals (e.g. O 2 .- ) oxidise Mito-SOX to fluorescent ethidium or 2-hydroxyethidium products whereas H 2 O 2 and/or peroxynitrite oxidise MitoPY1 to a fluorescent phenol by a boronate de-protection mechanism [41] , [42] , [43] , [44] , [45] , [46] , [47] . α-BTX increased Mito-SOX and MitoPY1 oxidation by 40.5 ± 4.9% and 30.5 ± 3.5%, respectively, compared to control ( Fig.…”

Section: Resultsmentioning

confidence: 99%

Mitochondrial ROS cause motor deficits induced by synaptic inactivity: Implications for synapse pruning

et al. 2018

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Developmental synapse pruning refines burgeoning connectomes. The basic mechanisms of mitochondrial reactive oxygen species (ROS) production suggest they select inactive synapses for pruning: whether they do so is unknown. To begin to unravel whether mitochondrial ROS regulate pruning, we made the local consequences of neuromuscular junction (NMJ) pruning detectable as motor deficits by using disparate exogenous and endogenous models to induce synaptic inactivity en masse in developing Xenopus laevis tadpoles. We resolved whether: (1) synaptic inactivity increases mitochondrial ROS; and (2) chemically heterogeneous antioxidants rescue synaptic inactivity induced motor deficits. Regardless of whether it was achieved with muscle (α-bungarotoxin), nerve (α-latrotoxin) targeted neurotoxins or an endogenous pruning cue (SPARC), synaptic inactivity increased mitochondrial ROS in vivo. The manganese porphyrins MnTE-2-PyP5+ and/or MnTnBuOE-2-PyP5+ blocked mitochondrial ROS to significantly reduce neurotoxin and endogenous pruning cue induced motor deficits. Selectively inducing mitochondrial ROS—using mitochondria-targeted Paraquat (MitoPQ)—recapitulated synaptic inactivity induced motor deficits; which were significantly reduced by blocking mitochondrial ROS with MnTnBuOE-2-PyP5+. We unveil mitochondrial ROS as synaptic activity sentinels that regulate the phenotypical consequences of forced synaptic inactivity at the NMJ. Our novel results are relevant to pruning because synaptic inactivity is one of its defining features.

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Section: Resultsmentioning

confidence: 99%

Mitochondrial ROS cause motor deficits induced by synaptic inactivity: Implications for synapse pruning

et al. 2018

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“…Optimal stimulation of murine macrophages leads to the extracellular formation of ∼0.1–0.2 nmol peroxynitrite min −1 /10 6 cells, which translates into a phagosomal peroxynitrite production of 0.1–0.4 mM s −1 , in the small T. cruzi –containing vacuole (3–5 fl; Alvarez et al, 2002). A recent study assessed peroxynitrite generation in response to phagocytosed T. cruzi , using a fluorescein-based boronate-derived probe that takes advantage of the fast reaction of peroxynitrite with boronic-esters (Zielonka et al, 2012; Rios et al, 2016; Prolo et al, 2018). The study showed that the amount of peroxynitrite needed to kill T. cruzi (LD 100 ) was ∼0.6 fmol/parasite; thus the amount of peroxynitrite generated during the time that the pathogen remains in the macrophage phagosome (90 min) is sufficient to kill the parasite.…”

Section: Kinetics and Toxicity Of Phagosomal Reactive Species In Macrmentioning

confidence: 99%

Reactive species and pathogen antioxidant networks during phagocytosis

Piacenza

Trujillo

Radí

2019

Journal of Experimental Medicine

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The generation of phagosomal cytotoxic reactive species (i.e., free radicals and oxidants) by activated macrophages and neutrophils is a crucial process for the control of intracellular pathogens. The chemical nature of these species, the reactions they are involved in, and the subsequent effects are multifaceted and depend on several host- and pathogen-derived factors that influence their production rates and catabolism inside the phagosome. Pathogens rely on an intricate and synergistic antioxidant armamentarium that ensures their own survival by detoxifying reactive species. In this review, we discuss the generation, kinetics, and toxicity of reactive species generated in phagocytes, with a focus on the response of macrophages to internalized pathogens and concentrating on Mycobacterium tuberculosis and Trypanosoma cruzi as examples of bacterial and parasitic infection, respectively. The ability of pathogens to deal with host-derived reactive species largely depends on the competence of their antioxidant networks at the onset of invasion, which in turn can tilt the balance toward pathogen survival, proliferation, and virulence over redox-dependent control of infection.

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“…The rate constant of the reaction, however, is significantly higher (~10 6 M -1 s -1 for ONOOand ~1 M -1 s -1 for H2O2), and the reaction typically involves a minor pathway, leading to ONOO -specific minor products (59). The high rate constant provides an opportunity to estimate the absolute flux of ONOOin cultured cells (48,60,61). Formation of ONOO --specific products provides an opportunity to selectively monitor ONOOformation in chemical and biological systems (14).…”

Section: Peroxynitritementioning

confidence: 99%

Tracking isotopically labeled oxidants using boronate-based redox probes

Ríos

Radí

Kalyanaraman

et al. 2020

Journal of Biological Chemistry

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Reactive oxygen and nitrogen species have been implicated in many biological processes and diseases, including immune responses, cardiovascular dysfunction, neurodegeneration, and cancer. These chemical species are short-lived in biological settings, and detecting them in these conditions and diseases requires the use of molecular probes that form stable, easily detectable, products. The chemical mechanisms and limitations of many of the currently used probes are not well-understood, hampering their effective applications. Boronates have emerged as a class of probes for the detection of nucleophilic two-electron oxidants. Here, we report the results of an oxygen-18–labeling MS study to identify the origin of oxygen atoms in the oxidation products of phenylboronate targeted to mitochondria. We demonstrate that boronate oxidation by hydrogen peroxide, peroxymonocarbonate, hypochlorite, or peroxynitrite involves the incorporation of oxygen atoms from these oxidants. We therefore conclude that boronates can be used as probes to track isotopically labeled oxidants. This suggests that the detection of specific products formed from these redox probes could enable precise identification of oxidants formed in biological systems. We discuss the implications of these results for understanding the mechanism of conversion of the boronate-based redox probes to oxidant-specific products.

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Fluorescence and chemiluminescence approaches for peroxynitrite detection

Cited by 81 publications

References 111 publications

Mitochondrial ROS cause motor deficits induced by synaptic inactivity: Implications for synapse pruning

Mitochondrial ROS cause motor deficits induced by synaptic inactivity: Implications for synapse pruning

Reactive species and pathogen antioxidant networks during phagocytosis

Tracking isotopically labeled oxidants using boronate-based redox probes

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