scholarly journals In Situ Detection and Measurement of Intracellular Reactive Oxygen Species in Single Isolated Mature Skeletal Muscle Fibers by Real Time Fluorescence Microscopy

2008 ◽  
Vol 10 (8) ◽  
pp. 1463-1474 ◽  
Author(s):  
Jesus Palomero ◽  
Deborah Pye ◽  
Tabitha Kabayo ◽  
David G. Spiller ◽  
Malcolm J. Jackson
Keyword(s):  
Reactive Oxygen Species ◽  
Skeletal Muscle ◽  
Fluorescence Microscopy ◽  
Real Time ◽  
Muscle Fibers ◽  
Reactive Oxygen ◽  
Intracellular Reactive Oxygen Species ◽  
Oxygen Species ◽  
In Situ Detection

scholarly journals Reactive oxygen species formation during tetanic contractions in single isolated Xenopus myofibers

2011 ◽  
Vol 111 (3) ◽  
pp. 898-904 ◽  
Author(s):  
Li Zuo ◽  
Leonardo Nogueira ◽  
Michael C. Hogan
Keyword(s):  
Reactive Oxygen Species ◽  
Skeletal Muscle ◽  
Real Time ◽  
Contractile Activity ◽  
Reactive Oxygen ◽  
Ros Generation ◽  
Oxygen Species ◽  
Fatigue Development ◽  
Species Formation ◽  
Significant Difference

Contracting skeletal muscle produces reactive oxygen species (ROS) that have been shown to affect muscle function and adaptation. However, real-time measurement of ROS in contracting myofibers has proven to be difficult. We used amphibian ( Xenopus laevis) muscle to test the hypothesis that ROS are formed during contractile activity in isolated single skeletal muscle fibers and that this contraction-induced ROS formation affects fatigue development. Single myofibers were loaded with 5 μM dihydrofluorescein-DA (Hfluor-DA), a fluorescent probe that reacts with ROS and results in the formation of fluorescein (Fluor) to precisely monitor ROS generation within single myofibers in real time using confocal miscroscopy. Three identical periods of maximal tetanic contractions (1 contraction/3 s for 2 min, separated by 60 min of rest) were conducted by each myofiber ( n = 6) at 20°C. Ebselen (an antioxidant) was present in the perfusate (10 μM) during the second contractile period. Force was reduced by ∼30% during each of the three contraction periods, with no significant difference in fatigue development among the three periods. The Fluor signal, indicative of ROS generation, increased significantly above baseline in both the first (42 ± 14%) and third periods (39 ± 10%), with no significant difference in the increase in fluorescence between the first and third periods. There was no increase of Fluor in the presence of ebselen during the second contractile period. These results demonstrated that, in isolated intact Xenopus myofibers, 1) ROS can be measured in real time during tetanic contractions, 2) contractile activity induced a significant increase above resting levels of ROS production, and 3) ebselen treatment reduced ROS generation to baseline levels but had no effect on myofiber contractility and fatigue development.

Activation of in situ tissue transglutaminase by intracellular reactive oxygen species

2003 ◽  
Vol 305 (3) ◽  
pp. 633-640 ◽  
Author(s):  
Zee-Won Lee ◽  
Sang-Mo Kwon ◽  
Sung-Woo Kim ◽  
Sun-Ju Yi ◽  
Young-Myeong Kim ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Tissue Transglutaminase ◽  
Reactive Oxygen ◽  
Intracellular Reactive Oxygen Species ◽  
Oxygen Species

scholarly journals Low Po2 conditions induce reactive oxygen species formation during contractions in single skeletal muscle fibers

2013 ◽  
Vol 304 (11) ◽  
pp. R1009-R1016 ◽  
Author(s):  
Li Zuo ◽  
Amy Shiah ◽  
William J. Roberts ◽  
Michael T. Chien ◽  
Peter D. Wagner ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Skeletal Muscle ◽  
Muscle Fibers ◽  
Reactive Oxygen ◽  
Rest Period ◽  
Convincing Evidence ◽  
Ros Generation ◽  
Oxygen Species ◽  
Skeletal Muscle Fibers ◽  
Ros Formation

Contractions in whole skeletal muscle during hypoxia are known to generate reactive oxygen species (ROS); however, identification of real-time ROS formation within isolated single skeletal muscle fibers has been challenging. Consequently, there is no convincing evidence showing increased ROS production in intact contracting fibers under low Po2 conditions. Therefore, we hypothesized that intracellular ROS generation in single contracting skeletal myofibers increases during low Po2 compared with a value approximating normal resting Po2. Dihydrofluorescein was loaded into single frog ( Xenopus) fibers, and fluorescence was used to monitor ROS using confocal microscopy. Myofibers were exposed to two maximal tetanic contractile periods (1 contraction/3 s for 2 min, separated by a 60-min rest period), each consisting of one of the following treatments: high Po2 (30 Torr), low Po2 (3–5 Torr), high Po2 with ebselen (antioxidant), or low Po2 with ebselen. Ebselen (10 μM) was administered before the designated contractile period. ROS formation during low Po2 treatment was greater than during high Po2 treatment, and ebselen decreased ROS generation in both low- and high-Po2 conditions ( P < 0.05). ROS accumulated at a faster rate in low vs. high Po2. Force was reduced >30% for each condition except low Po2 with ebselen, which only decreased ∼15%. We concluded that single myofibers under low Po2 conditions develop accelerated and more oxidative stress than at Po2 = 30 Torr (normal human resting Po2). Ebselen decreases ROS formation in both low and high Po2, but only mitigates skeletal muscle fatigue during reduced Po2 conditions.

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