Abstract
Objective:Oxidative stress plays an important role in the onset of many neuronal and peripheral disorders. We examined the feasibility of obtaining semiquantitative fluorescent images of reactive oxygen species (ROS) generation in mouse brain and kidney utilizing a planar laser scanner and dihydroethidium (DHE).Methods:To investigate ROS generation in brain, sodium nitroprusside was injected into the striatum. Dihydroethidium was injected into the tail vein. After DHE injection, tissue slices were analyzed utilizing a planar laser scanner. For kidney study, cis-diamminedichloroplatinum [II] (cisplatin) was intraperitoneally administrated into mice.Results:Clear and semiquantitative fluorescent images of ROS generation in the mouse brain and kidney were obtained. Furthermore, the fluorescence intensity was stable and not affected by fading. Sodium nitroprusside induced approximately 6 times the fluorescence accumulation in the brain. Cisplatin caused renal injury in all mice, and in comparison with control mice, more than 10 times fluorescence accumulation was observed in the renal medulla with tubular necrosis and vacuolization.Conclusions:We successfully obtained ex vivo semiquantitative fluorescent images of ROS generation utilizing a planar laser scanner and DHE. This simple method is useful for ROS detection in several ROS-related animal models and would be applicable to a variety of biochemical processes.
Highlights
Reactive oxygen species (ROS) play an important role as regulatory mediators in signaling processes under physiological conditions, and relatively low ROS concentrations are maintained by the redox regulation system.[1]
We examined a simple procedure for imaging the ROS generation in intact animals utilizing a planar laser scanner, Fluoro-Imaging Analyzer System (FLA-7000; Fuji Film Co, Tokyo, Japan) and a fluorescent probe, dihydroethidium (DHE)
The result indicated that the fluorescence intensity is stable and not affected by the fading, which often occurs during microscopic observation
Summary
Reactive oxygen species (ROS) play an important role as regulatory mediators in signaling processes under physiological conditions, and relatively low ROS concentrations are maintained by the redox regulation system.[1] the persistent production of excessive amounts of ROS induces a disturbance in redox homeostasis. ROS are implicated in the onset of many neurodegenerative disorders and the development of adverse renal and/or cardiac side effects for cancer chemotherapy. The development of the method for in vivo ROS imaging is an important subject for both basic and clinical research.
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
