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A5494 - A Far Red Emitting Fluorescence Probe Detect Specific Peroxynitrite In Vitro and Is Used for Detection of Peroxynitrite in Live Cells and Lung Infectious Mouse Model
Author Block: D. Jeong1, J. Ryu2, Y. Chung3, J. Yoon4, J. Yoon5, J. Ryu2; 1Severance Biomedical Science Institute, Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul 120-752, Korea., Seoul, Korea, Republic of, Seoul, Korea, Republic of, 2Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul, Korea, Seoul, Korea, Republic of, 3The Airway Mucus Institute, Yonsei University College of Medicine, Seoul, Korea, Seoul, Korea, Republic of, 4Department of Otorhinolaryngology, Yonsei University College of Medicine, Seoul, Korea, Seoul, Korea, Republic of, 5Department of Chemistry and Nano Science, Ewha Womans University, Seoul, 120-750, Korea, Seoul, Korea, Republic of.
It is well known that reactive oxygen species (ROS) and reactive nitrogen species (RNS) are related to various pathophysiological processes. Among ROS and RNS, peroxynitrite (ONOO−) has attracted much attention because of its key roles in signal transduction and antimicrobial activities. Peroxynitrite is produced by the reaction of superoxide anion (O2−•) with nitric oxide (•NO). When Peroxynitrite is mis-regulated, it is regarded as a key contributor to numerous pathologies such as cancer, diabetes, and cardiovascular disease. Thus, it is essential to know the peroxynitrite’s diverse roles in pathophysiological functions. But in living system, ONOO− has a so short lifetime (about 10 ms) that it is impossible to directly measure ONOO− in living cell or tissue samples by using established methods. In the previous study, many fluorescence probes which detect peroxynitrite were reported and these probes have a variety of reactive functional group. On the basis of recent study, we expect that probe using hydrazide among reactive functional group can detect specific ONOO-. So, to develop ONOO− detection and imaging, we found that the far red emitting fluorescence probe 1, containing a rhodamine derivative and a hydrazide reactive group. We experimented in HeLa and RAW 264.7 cells whether this probe can be utilized or not. In addition, we isolated bone marrow-derived neutrophils in lung infected mouse with GFP-tagged Pseudomonas aeruginosa (PAO1) and detected peroxynitrite by far red emitting fluorescence probe 1. This experiments revels that this probe can detect exogenous and endogenous peroxynitrite in pathogen-infected mouse model system. Consequently, this probe has cell permeability and can detect more sensitive and selective fluorescence of peroxynitrite than other ROS/RNS. Moreover, long wavelength absorption (600 nm) and emission (638 nm) bands occur when this probe reacts with peroxynitrite. This characteristic has some advantages such as deep photon penetration in tissue, minimum photo-damage to biomolecules and minimum interference from background auto-fluorescence of biological samples. We expect that a far red emitting fluorescence probe will utilize for molecular imaging in observations of the roles played by peroxynitrite in pathophysiological processes.