scholarly journals Real-time imaging of hydrogen peroxide dynamics in vegetative and pathogenic hyphae of Fusarium graminearum

2015 ◽  
Vol 5 (1) ◽  
Author(s):  
Michael Mentges ◽  
Jörg Bormann
Keyword(s):  
Hydrogen Peroxide ◽  
Real Time ◽  
Fusarium Graminearum ◽  
Real Time Imaging

scholarly journals Real-time imaging of intracellular hydrogen peroxide in pancreatic islets

2016 ◽  
Vol 473 (23) ◽  
pp. 4443-4456 ◽  
Author(s):  
Adam Neal ◽  
Austin Rountree ◽  
Kelly Kernan ◽  
Brian Van Yserloo ◽  
Huiliang Zhang ◽  
...  
Keyword(s):  
Hydrogen Peroxide ◽  
Real Time ◽  
Live Cells ◽  
H2o2 Production ◽  
Spatiotemporal Resolution ◽  
Intact Cells ◽  
Real Time Imaging ◽  
Mitochondrial Superoxide ◽  
Acute Changes ◽  
Intracellular Hydrogen Peroxide

A real-time method to measure intracellular hydrogen peroxide (H2O2) would be very impactful in characterizing rapid changes that occur in physiologic and pathophysiologic states. Current methods do not provide the sensitivity, specificity and spatiotemporal resolution needed for such experiments on intact cells. We developed the use of HyPer, a genetic indicator for H2O2 that can be expressed in the cytosol (cyto-HyPer) or the mitochondria (mito-HyPer) of live cells. INS-1 cells or islets were permeabilized and the cytosolic HyPer signal was a linear function of extracellular H2O2, allowing fluorescent cyto-HyPer signals to be converted into H2O2 concentrations. Glucose increased cytosolic H2O2, an effect that was suppressed by overexpression of catalase. Large perturbations in pH can influence the HyPer signal, but inclusion of HEPES [4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid] in the perfusate prevented pH changes, but did not affect glucose-induced cyto-HyPer signals, suggesting that this effect is largely pH-independent. Using the assay, two fundamental questions were addressed. Knockdown of superoxide dismutase 2 (SOD2), the mitochondrial form of SOD, completely suppressed glucose-induced H2O2. Furthermore, glucose also induced mitochondrial superoxide and H2O2 production, which preceded the appearance of cytosolic H2O2. Therefore, glucose-induced H2O2 largely originated from mitochondria. Finally, the glucose-induced HyPer signal was less than 1/20th of that induced by toxic levels of H2O2. Overall, the use of HyPer for real-time imaging allowed resolution of acute changes in intracellular levels of H2O2 and will have great utility for islet studies involving mechanisms of H2O2-mediated signaling and oxidative stress.

Fluorogenic Probe Using a Mislow–Evans Rearrangement for Real‐Time Imaging of Hydrogen Peroxide

2020 ◽  
Vol 132 (40) ◽  
pp. 17588-17594
Author(s):  
Dianne Pham ◽  
Upamanyu Basu ◽  
Ivanna Pohorilets ◽  
Claudette M. St Croix ◽  
Simon C. Watkins ◽  
...  
Keyword(s):  
Hydrogen Peroxide ◽  
Real Time ◽  
Real Time Imaging ◽  
Fluorogenic Probe

Real-Time Imaging of Mitochondrial Hydrogen Peroxide and pH Fluctuations in Living Cells Using a Fluorescent Nanosensor

2015 ◽  
Vol 87 (7) ◽  
pp. 3678-3684 ◽  
Author(s):  
Limin Yang ◽  
Na Li ◽  
Wei Pan ◽  
Zhengze Yu ◽  
Bo Tang
Keyword(s):  
Hydrogen Peroxide ◽  
Real Time ◽  
Living Cells ◽  
Real Time Imaging ◽  
Ph Fluctuations

scholarly journals Fluorogenic Probe Using a Mislow-Evans Rearrangement for Real-Time Imaging of Hydrogen Peroxide

2020 ◽  
Author(s):  
Dianne Pham ◽  
Upamanyu Basu ◽  
Ivanna Pohorilets ◽  
Claudette M. St Croix ◽  
Simon Watkins ◽  
...  
Keyword(s):  
Hydrogen Peroxide ◽  
Real Time ◽  
Small Molecule ◽  
Sigmatropic Rearrangement ◽  
Probe Design ◽  
Fast Kinetics ◽  
Real Time Imaging ◽  
Fluorogenic Probe ◽  
Fluorogenic Probes ◽  
Green Fluorescent

<p>Hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>) is a second messenger that mediates the biology of wound healing, apoptosis, inflammation, aging, neurodegenerative diseases, and more. Its presence has been fluorometrically imaged with protein- or small molecule-based sensors. However, only protein-based sensors have afforded temporal insights with the resolution of seconds. Small molecule-based fluorogenic probes are preferred for various reasons; however, current electrophilic chemosensors react with H<sub>2</sub>O<sub>2</sub> slowly, requiring >20 minutes for a sufficient response. Here, we report a fluorogenic probe that selectively reacts with H<sub>2</sub>O<sub>2</sub> and undergoes a [2,3]-sigmatropic rearrangement (seleno-Mislow-Evans rearrangement) followed by an acetal hydrolysis to produce a green fluorescent molecule in seconds. The mode of reaction is based on the umpolung of previously developed sensors; the probe acts as a nucleophile rather than an electrophile. The fast kinetics outcompete the reaction between thiols and H<sub>2</sub>O<sub>2</sub>, enabling real-time imaging of H<sub>2</sub>O<sub>2</sub> produced inside the subcellular compartments of cells in 8 seconds. Further, the probe was able to recapitulate data previously observed only with a genetically encoded protein-based sensor. The present probe design provides a platform that can match the temporal resolution of protein-based H<sub>2</sub>O<sub>2</sub> detection. </p>

A near-infrared fluorescent probe for endogenous hydrogen peroxide real-time imaging in living cells and zebrafish

2019 ◽  
Vol 165 ◽  
pp. 518-523 ◽  
Author(s):  
Xin Huang ◽  
Zhipeng Li ◽  
Zixin Liu ◽  
Chengchu Zeng ◽  
Liming Hu
Keyword(s):  
Hydrogen Peroxide ◽  
Real Time ◽  
Fluorescent Probe ◽  
Near Infrared ◽  
Living Cells ◽  
Real Time Imaging

scholarly journals Fluorogenic Probe Using a Mislow-Evans Rearrangement for Real-Time Imaging of Hydrogen Peroxide

2020 ◽  
Author(s):  
Dianne Pham ◽  
Upamanyu Basu ◽  
Ivanna Pohorilets ◽  
Claudette M. St Croix ◽  
Simon Watkins ◽  
...  
Keyword(s):  
Hydrogen Peroxide ◽  
Real Time ◽  
Small Molecule ◽  
Sigmatropic Rearrangement ◽  
Probe Design ◽  
Fast Kinetics ◽  
Real Time Imaging ◽  
Fluorogenic Probe ◽  
Fluorogenic Probes ◽  
Green Fluorescent

<p>Hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>) is a second messenger that mediates the biology of wound healing, apoptosis, inflammation, aging, neurodegenerative diseases, and more. Its presence has been fluorometrically imaged with protein- or small molecule-based sensors. However, only protein-based sensors have afforded temporal insights with the resolution of seconds. Small molecule-based fluorogenic probes are preferred for various reasons; however, current electrophilic chemosensors react with H<sub>2</sub>O<sub>2</sub> slowly, requiring >20 minutes for a sufficient response. Here, we report a fluorogenic probe that selectively reacts with H<sub>2</sub>O<sub>2</sub> and undergoes a [2,3]-sigmatropic rearrangement (seleno-Mislow-Evans rearrangement) followed by an acetal hydrolysis to produce a green fluorescent molecule in seconds. The mode of reaction is based on the umpolung of previously developed sensors; the probe acts as a nucleophile rather than an electrophile. The fast kinetics outcompete the reaction between thiols and H<sub>2</sub>O<sub>2</sub>, enabling real-time imaging of H<sub>2</sub>O<sub>2</sub> produced inside the subcellular compartments of cells in 8 seconds. Further, the probe was able to recapitulate data previously observed only with a genetically encoded protein-based sensor. The present probe design provides a platform that can match the temporal resolution of protein-based H<sub>2</sub>O<sub>2</sub> detection. </p>

scholarly journals Fluorogenic Probe Using a Mislow–Evans Rearrangement for Real‐Time Imaging of Hydrogen Peroxide

2020 ◽  
Vol 59 (40) ◽  
pp. 17435-17441 ◽  
Author(s):  
Dianne Pham ◽  
Upamanyu Basu ◽  
Ivanna Pohorilets ◽  
Claudette M. St Croix ◽  
Simon C. Watkins ◽  
...  
Keyword(s):  
Hydrogen Peroxide ◽  
Real Time ◽  
Real Time Imaging ◽  
Fluorogenic Probe
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