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Título
Genetically Encoded Biosensors to Monitor Intracellular Reactive Oxygen and Nitrogen Species and Glutathione Redox Potential in Skeletal Muscle Cells
Autor(es)
Materia
Biosensors
Quantitative fluorescence microscopy
Redox signaling
RONS
Hydrogen peroxide
Nitric oxide
Glutathione
Redox potential
Skeletal muscle
C2C12
Myoblast/myotube
Single skeletal muscle fiber
Clasificación UNESCO
2411.10 Fisiología del Músculo
Fecha de publicación
2021
Editor
Board
Citación
Fernández Puente, E., Palomero Labajos, J. (2021). Genetically Encoded Biosensors to Monitor Intracellular Reactive Oxygen and Nitrogen Species and Glutathione Redox Potential in Skeletal Muscle Cells. International Journal of Molecular Sciences, 22 (19) pp 1-20. https://doi.org/10.3390/ijms221910876
Resumen
[EN] Reactive oxygen and nitrogen species (RONS) play an important role in the pathophysiology of skeletal muscle and are involved in the regulation of intracellular signaling pathways,
which drive metabolism, regeneration, and adaptation in skeletal muscle. However, the molecular
mechanisms underlying these processes are unknown or partially uncovered. We implemented a
combination of methodological approaches that are funded for the use of genetically encoded biosensors associated with quantitative fluorescence microscopy imaging to study redox biology in skeletal
muscle. Therefore, it was possible to detect and monitor RONS and glutathione redox potential with
high specificity and spatio-temporal resolution in two models, isolated skeletal muscle fibers and
C2C12 myoblasts/myotubes. Biosensors HyPer3 and roGFP2-Orp1 were examined for the detection
of cytosolic hydrogen peroxide; HyPer-mito and HyPer-nuc for the detection of mitochondrial and
nuclear hydrogen peroxide; Mito-Grx1-roGFP2 and cyto-Grx1-roGFP2 were used for registration
of the glutathione redox potential in mitochondria and cytosol. G-geNOp was proven to detect
cytosolic nitric oxide. The fluorescence emitted by the biosensors is affected by pH, and this might
have masked the results; therefore, environmental CO2 must be controlled to avoid pH fluctuations.
In conclusion, genetically encoded biosensors and quantitative fluorescence microscopy provide
a robust methodology to investigate the pathophysiological processes associated with the redox
biology of skeletal muscle.
URI
DOI
10.3390/IJMS221910876
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