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Observation of the Generation of ONOO(-) in Mitochondria under Various Stimuli with a Sensitive Fluorescence Probe.
Analytical Chemistry 2017 May 3
Peroxynitrite (ONOO(-)), an important reactive oxygen species (ROS), may be generated in the mitochondria under various physiological and pathological conditions. In this work, we designed and synthesized a mitochondria-targetable fluorescence probe RPTPP by using rhodamine as the fluorophore, phenylhydrazine as the recognition moiety, and triphenyl-phosphonium cation as the mitochondria-targeting moiety. Upon reaction of the probe with ONOO(-), the oxidation of phenylhydrazine by ONOO(-) and the subsequent hydrolysis opens the nonfluorescent spirocyclic structure and, thus, triggers a fluorescence turn-on response, which provides a sensitive and selective method for the detection of ONOO(-). The mitochondria-targeting property of RPTPP was confirmed by the colocalization experiments as well as the mitochondria uncoupling treatments. Moreover, the applications of the probe for imaging intracellular ONOO(-) were performed in living cells, which reveals that the ONOO(-) level in RAW264.7 cells undergoes an about 9-fold increase with the stimulation of LPS and IFN-γ for 15 h.
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