Detection of Mitochondrial Reactive Oxygen Species in Living Rat Trigeminal Caudal Neurons
International Journal of Oral Biology
; : 103-109, 2015.
Article
in Ko
| WPRIM
| ID: wpr-104522
Responsible library:
WPRO
ABSTRACT
Growing evidence suggests that mitochondrial reactive oxygen species (ROS) are involved in various pain states. This study was performed to investigate whether ROS-induced changes in neuronal excitability in trigeminal subnucleus caudalis are related to ROS generation in mitochondria. Confocal scanning laser microscopy was used to measure ROS-induced fluorescence intensity in live rat trigeminal caudalis slices. The ROS level increased during the perfusion of malate, a mitochondrial substrate, after loading of 2',7'-dichlorofluorescin diacetate (H2DCF-DA), an indicator of the intracellular ROS; the ROS level recovered to the control condition after washout. When pre-treated with phenyl N-tert-butylnitrone (PBN) and 4-hydroxy-2,2,6,6-tetramethylpiperidene-1-oxyl (TEMPOL), malate-induced increase of ROS level was suppressed. To identify the direct relation between elevated ROS levels and mitochondria, we applied the malate after double-loading of H2DCF-DA and chloromethyl-X-rosamine (CMXRos; MitoTracker Red), which is a mitochondria-specific fluorescent probe. As a result, increase of both intracellular ROS and mitochondrial ROS were observed simultaneously. This study demonstrated that elevated ROS in trigeminal subnucleus caudalis neuron can be induced through mitochondrial-ROS pathway, primarily by the leakage of ROS from the mitochondrial electron transport chain.
Key words
Full text:
1
Index:
WPRIM
Main subject:
Perfusion
/
Reactive Oxygen Species
/
Microscopy, Confocal
/
Electron Transport
/
Fluorescence
/
Mitochondria
/
Neurons
Type of study:
Diagnostic_studies
Limits:
Animals
Language:
Ko
Journal:
International Journal of Oral Biology
Year:
2015
Type:
Article