Abstract:Mitochondrial flashes (mitoflashes) are recently discovered mitochondrial activity which reflects chemical and electrical excitation of the organelle. Emerging evidence indicates that mitoflashes represent highly regulated, elementary signaling events that play important roles in physiological and pathophysiological processes in eukaryotes. Furthermore, they are regulated by mitochondrial ROS, Ca, and protons, and are intertwined with mitochondrial metabolic processes. As such, targeting mitoflash activity may… Show more
“…Superoxide flashes are 10-s events that occur spontaneously and suddenly in mitochondria and reflect electrical and chemical activities (Feng et al 2017). Superoxide flashes were first defined as transient events of the mitochondrial matrix-targeted biosensor mt-cp YFP (Wei and PeerJ reviewing PDF | (2018:11:33136:1:1:NEW 22 Feb 2019)…”
Section: Introductionmentioning
confidence: 99%
“…Manuscript to be reviewed Dirksen 2012). As mt-cp YFP is sensitive to pH, superoxide flashes can be visualized by chemical probes, including ROS indicators, such as MitoSOX for superoxide and 2.7dichlorodihyfrofluorescein (DCF) for H 2 O 2 (Feng et al 2017;Zhang et al 2013). Interestingly, cp-YFP superoxide flashes are correlated with depolarization of the mitochondrial membrane potential (ΔΨ m ) (Zhang et al 2013).…”
Background Mitochondria are the center of energy metabolism and the production of reactive oxygen species (ROS). ROS production results in a burst of "superoxide flashes", which is always accompanied by depolarization of mitochondrial membrane potential. Superoxide flashes have only been studied in the model plant Arabidopsis thaliana using a complex method to isolate mitochondria. In this study, we present an efficient, easier method to isolate functional mitochondria from floral tissues to measure superoxide flashes. Method We used 0.5 g samples to isolate mitochondria within < 1.5 h from flowers of two nontransgenic plants (Magnolia denudata and Nelumbo nucifera) to measure superoxide flashes. Superoxide flashes were visualized by the pH-insensitive indicator MitoSOX Red, while the mitochondrial membrane potential (ΔΨm) was labelled with TMRM. Results Mitochondria isolated using our method showed a high respiration ratio. Our results indicate that the location of ROS and mitochondria was in a good coincidence. Increased ROS together with a higher frequency of superoxide flashes was found in mitochondria isolated from the flower pistil. Furthermore, a higher rate of depolarization of the ΔΨm was observed in the pistil. Taken together, these results demonstrate that the frequency of superoxide flashes is closely related to depolarization of the ΔΨm in petals and pistils of flowers.
“…Superoxide flashes are 10-s events that occur spontaneously and suddenly in mitochondria and reflect electrical and chemical activities (Feng et al 2017). Superoxide flashes were first defined as transient events of the mitochondrial matrix-targeted biosensor mt-cp YFP (Wei and PeerJ reviewing PDF | (2018:11:33136:1:1:NEW 22 Feb 2019)…”
Section: Introductionmentioning
confidence: 99%
“…Manuscript to be reviewed Dirksen 2012). As mt-cp YFP is sensitive to pH, superoxide flashes can be visualized by chemical probes, including ROS indicators, such as MitoSOX for superoxide and 2.7dichlorodihyfrofluorescein (DCF) for H 2 O 2 (Feng et al 2017;Zhang et al 2013). Interestingly, cp-YFP superoxide flashes are correlated with depolarization of the mitochondrial membrane potential (ΔΨ m ) (Zhang et al 2013).…”
Background Mitochondria are the center of energy metabolism and the production of reactive oxygen species (ROS). ROS production results in a burst of "superoxide flashes", which is always accompanied by depolarization of mitochondrial membrane potential. Superoxide flashes have only been studied in the model plant Arabidopsis thaliana using a complex method to isolate mitochondria. In this study, we present an efficient, easier method to isolate functional mitochondria from floral tissues to measure superoxide flashes. Method We used 0.5 g samples to isolate mitochondria within < 1.5 h from flowers of two nontransgenic plants (Magnolia denudata and Nelumbo nucifera) to measure superoxide flashes. Superoxide flashes were visualized by the pH-insensitive indicator MitoSOX Red, while the mitochondrial membrane potential (ΔΨm) was labelled with TMRM. Results Mitochondria isolated using our method showed a high respiration ratio. Our results indicate that the location of ROS and mitochondria was in a good coincidence. Increased ROS together with a higher frequency of superoxide flashes was found in mitochondria isolated from the flower pistil. Furthermore, a higher rate of depolarization of the ΔΨm was observed in the pistil. Taken together, these results demonstrate that the frequency of superoxide flashes is closely related to depolarization of the ΔΨm in petals and pistils of flowers.
“…Superoxide flashes are 10-s events that occur spontaneously and suddenly in mitochondria and reflect electrical and chemical activities (Feng et al, 2017). Superoxide flashes were first defined as transient events of the mitochondrial matrix-targeted biosensor mt-cp YFP (Wei & Dirksen, 2012).…”
Section: Introductionmentioning
confidence: 99%
“…Superoxide flashes were first defined as transient events of the mitochondrial matrix-targeted biosensor mt-cp YFP (Wei & Dirksen, 2012). As mt-cp YFP is sensitive to pH, superoxide flashes can be visualized by chemical probes, including ROS indicators, such as MitoSOX for superoxide and 2.7-dichlorodihyfrofluorescein (DCF) for H 2 O 2 (Feng et al, 2017; Zhang et al, 2013). Interestingly, cp-YFP superoxide flashes are correlated with depolarization of the mitochondrial membrane potential (ΔΨ m ) (Zhang et al, 2013).…”
Background
Mitochondria are the center of energy metabolism and the production of reactive oxygen species (ROS). ROS production results in a burst of “superoxide flashes”, which is always accompanied by depolarization of mitochondrial membrane potential. Superoxide flashes have only been studied in the model plant Arabidopsis thaliana using a complex method to isolate mitochondria. In this study, we present an efficient, easier method to isolate functional mitochondria from floral tissues to measure superoxide flashes.
Method
We used 0.5 g samples to isolate mitochondria within <1.5 h from flowers of two non-transgenic plants (Magnolia denudata and Nelumbo nucifera) to measure superoxide flashes. Superoxide flashes were visualized by the pH-insensitive indicator MitoSOX Red, while the mitochondrial membrane potential (ΔΨ m) was labelled with TMRM.
Results
Mitochondria isolated using our method showed a high respiration ratio. Our results indicate that the location of ROS and mitochondria was in a good coincidence. Increased ROS together with a higher frequency of superoxide flashes was found in mitochondria isolated from the flower pistil. Furthermore, a higher rate of depolarization of the ΔΨ m was observed in the pistil. Taken together, these results demonstrate that the frequency of superoxide flashes is closely related to depolarization of the ΔΨ m in petals and pistils of flowers.
“…Mitoflash has also been ubiquitously observed in diverse biological systems such as isolated mitochondria, cells in culture dishes, explanted organs, and enormous models ranging from worms and zebrafish to rodents and humans. 1,[5][6][7][8] Thus it was suggested as an elemental signaling event 2,9 in a variety of physiological and pathophysiological conditions.…”
Mitochondrial oxidative flashes (mitoflashes) are oxidative burst events in mitochondria. It is crosslinked with numerous mitochondrial molecular processes and related with pivotal cell functions such as apoptosis and respiration. In previous research, mitoflashes were found as spontaneous occasional events. It would be observed more frequently if cells were treated with proapoptotic chemicals. We show that multiple mitoflashes can be initiated by a single femtosecond-laser stimulation that was tightly focused on a diffraction-limited spot in the mitochondrial tubular structure. The mitoflash events triggered by different photostimulations are quantified and analyzed. The width and amplitude of mitoflashes are found very sensitive to stimulation parameters including laser power, exposure duration, and total incident laser energy. This study provides a quantitative investigation on the photostimulated mitoflashes. It may thus demonstrate such optical method to be a promising technique in future mitochondrial research.
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