Specific Visualization of Nitric Oxide in the Vasculature with Two-Photon Microscopy Using a Copper Based Fluorescent Probe

Ghosh, Mitrajit; Akker, Nynke M. S. van den; Wijnands, Karolina A. P.; Poeze, Martijn; Weber, Christian; McQuade, Lindsey E.; Pluth, Michael D.; Lippard, Stephen J.; Post, Mark J.; Molin, Daniël; Zandvoort, Marc A. M. J. van

doi:10.1371/journal.pone.0075331

Cited by 18 publications

(28 citation statements)

References 27 publications

(67 reference statements)

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“…Ex vivo NO production in endothelial cells was detected with Cu 2 Fl 2 using 2-photon imaging as described in the online Supplementary Materials and Method section in File S1 [39].…”

Section: Methodsmentioning

confidence: 99%

Arginase-1 Deficiency Regulates Arginine Concentrations and NOS2-Mediated NO Production during Endotoxemia

et al. 2014

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Rationale and objectiveArginase-1 is an important component of the intricate mechanism regulating arginine availability during immune responses and nitric oxide synthase (NOS) activity. In this study Arg1fl/fl/Tie2-Cretg/− mice were developed to investigate the effect of arginase-1 related arginine depletion on NOS2- and NOS3-dependent NO production and jejunal microcirculation under resting and endotoxemic conditions, in mice lacking arginase-1 in endothelial and hematopoietic cells.Methods and ResultsArginase-1-deficient mice as compared with control mice exhibited higher plasma arginine concentration concomitant with enhanced NO production in endothelial cells and jejunal tissue during endotoxemia. In parallel, impaired jejunal microcirculation was observed in endotoxemic conditions. Cultured bone-marrow-derived macrophages of arginase-1 deficient animals also presented a higher inflammatory response to endotoxin than control littermates. Since NOS2 competes with arginase for their common substrate arginine during endotoxemia, Nos2 deficient mice were also studied under endotoxemic conditions. As Nos2−/− macrophages showed an impaired inflammatory response to endotoxin compared to wild-type macrophages, NOS2 is potentially involved. A strongly reduced NO production in Arg1fl/fl/Tie2-Cretg/− mice following infusion of the NOS2 inhibitor 1400W further implicated NOS2 in the enhanced capacity to produce NO production Arg1fl/fl/Tie2-Cretg/− mice.ConclusionsReduced arginase-1 activity in Arg1fl/fl/Tie2-Cretg/− mice resulted in increased inflammatory response and NO production by NOS2, accompanied by a depressed microcirculatory flow during endotoxemia. Thus, arginase-1 deficiency facilitates a NOS2-mediated pro-inflammatory activity at the expense of NOS3-mediated endothelial relaxation.

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“…Ex vivo NO production in endothelial cells was detected with Cu 2 Fl 2 using 2-photon imaging as described in the online Supplementary Materials and Method section in File S1 [39].…”

Section: Methodsmentioning

confidence: 99%

Arginase-1 Deficiency Regulates Arginine Concentrations and NOS2-Mediated NO Production during Endotoxemia

et al. 2014

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“…The small size, volatility, short half-life (approximately 2 ms) 5 and other physical properties of NO present considerable challenges in developing reliable methods for its detection and accurate measurement within blood, cells and tissues. Many fluorescence-based sensors including diaminofluorescein 6,7 , BODIPY 8 , Near Infra-Red fluorescence 9–12 , carbon-nanotube 9,10 and metal-based turn-on fluorescent probes 13,14 have been developed to detect NO in cells, tissues and organs 15,16 . Electrochemical methods have been applied for NO sensing, leading to the development of many chemical multimodality sensors that have significant limitations based on their physical and chemical properties and toxicological profiles 17–19 .…”

Section: Introductionmentioning

confidence: 99%

A Novel Ruthenium-based Molecular Sensor to Detect Endothelial Nitric Oxide

Vidanapathirana

Pullen

Zhang

et al. 2019

Sci Rep

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Nitric oxide (NO) is a key regulator of endothelial cell and vascular function. The direct measurement of NO is challenging due to its short half-life, and as such surrogate measurements are typically used to approximate its relative concentrations. Here we demonstrate that ruthenium-based [Ru(bpy)2(dabpy)]2+ is a potent sensor for NO in its irreversible, NO-bound active form, [Ru(bpy)2(T-bpy)]2+. Using spectrophotometry we established the sensor’s ability to detect and measure soluble NO in a concentration-dependent manner in cell-free media. Endothelial cells cultured with acetylcholine or hydrogen peroxide to induce endogenous NO production showed modest increases of 7.3 ± 7.1% and 36.3 ± 25.0% respectively in fluorescence signal from baseline state, while addition of exogenous NO increased their fluorescence by 5.2-fold. The changes in fluorescence signal were proportionate and comparable against conventional NO assays. Rabbit blood samples immediately exposed to [Ru(bpy)2(dabpy)]2+ displayed 8-fold higher mean fluorescence, relative to blood without sensor. Approximately 14% of the observed signal was NO/NO adduct-specific. Optimal readings were obtained when sensor was added to freshly collected blood, remaining stable during subsequent freeze-thaw cycles. Clinical studies are now required to test the utility of [Ru(bpy)2(dabpy)]2+ as a sensor to detect changes in NO from human blood samples in cardiovascular health and disease.

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“…To further exploit the probes in living systems, these authors introduced one of the Cu-FL probes, Cu 2 FL 2 E(Fig. 7, complex 35), to visualize NO in vasculature with two-photon microscopy[103]. Porcine aortic cells (SMCs) and segments of murine common carotid arteries (tissue)…”

mentioning

confidence: 99%